RU2013153802A - Композиции и способы, включающие варианты сериновой протеазы - Google Patents
Композиции и способы, включающие варианты сериновой протеазы Download PDFInfo
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Abstract
1. Выделенный вариант субтилизина, где указанный вариант субтилизина представляет собой зрелую форму, обладающую протеолитической активностью, и содержит аминокислотную последовательность, включающую комбинацию замен аминокислот в положениях, выбранных из: T022R+V104I+Q245R, G020R+S024R+S101A, T022R+S103A+V104I, T022R+V104I+A232V, T022R+S103A+Q245R, T022R+A232V+Q245R, T022R+S103A+A232V, T022R+S103A+V104I+A232V, T022R+V104I+A232V+Q245R, T022R+S103A+V104I+Q245R, T022R+S103A+A232V+Q245R, T022R+S103A+V104I+A232V+Q245R, T022A+N076D+S103A+V104I+A232V+Q245R, T022A+N076D+S103A+V104I+A232V, T022A+N076D+V104I+A232V+Q245R, T022A+N076D+S103A+A232V+Q245R, T022A+N076D+S103A+V104I+Q245R, T022A+N076D+V104I+Q245R, T022A+N076D+A232V+Q245R, T022A+N076D+V104I+A232V, T022A+N076D+S103A+Q245R, T022A+N076D+S103A+A232V, T022A+N076D+S103A+V104I, T022A+N076D+S103A, T022A+N076D+V104I, T022A+N076D+A232V, T022A+N076D+Q245R, G020R+S101A+T213A, G020R+R045T+S101A, G020R+S101A+G211Q, S024R+S101A+T213A, G020R+T022W+S101A, S024R+S101A+G211Q, N043R+S101A+T213A, N043R+R045T+S101A, S024R+N043R+S101A, S024R+R045T+S101A, S101A+G211Q+T213A, G020R+N043R+S101A, R045T+S101A+T213A, T022W+S024R+S101A, T022R+S101G+V104I, T022W+S101A+T213A, T022R+S101G+Q245R, T022W+S101A+G211Q, G020R+S024R+Q245R, T022R+S101G+V104I+Q245R, T022R+S101G+A232V, T022R+S101G+S103A, T022R+S101G+S103A+V104I, T022R+S101G+S103A+Q245R, T022R+S101G+A232V+Q245R, T022R+S101G+V104I+A232V, G020R+S024R+S101A+T213A, G020R+S024R+T213AS101N+V104I+A232V, S101N+S103A+V104I, S101N+S103A+A232V, S101N+S103A+V104I+A232V, G020R+S103A+A232V, G020R+V104I+A232V, G020R+S103A+V104I+A232V, G020R+S103A+V104I, N018R+N076D+N116A, N018R+N076D+T213A, N018R+N076D+S101A, N018R+N076D+A215F или N018R+N076D+G211Q, и где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.2. Выделенный вариант субтилизина по п. 1, где указанными положениями являются T022A+N076D+S103A+V104I+A232V+Q245R, и где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.3. Выделенный вариант субтилизина по п. 1
Claims (52)
1. Выделенный вариант субтилизина, где указанный вариант субтилизина представляет собой зрелую форму, обладающую протеолитической активностью, и содержит аминокислотную последовательность, включающую комбинацию замен аминокислот в положениях, выбранных из: T022R+V104I+Q245R, G020R+S024R+S101A, T022R+S103A+V104I, T022R+V104I+A232V, T022R+S103A+Q245R, T022R+A232V+Q245R, T022R+S103A+A232V, T022R+S103A+V104I+A232V, T022R+V104I+A232V+Q245R, T022R+S103A+V104I+Q245R, T022R+S103A+A232V+Q245R, T022R+S103A+V104I+A232V+Q245R, T022A+N076D+S103A+V104I+A232V+Q245R, T022A+N076D+S103A+V104I+A232V, T022A+N076D+V104I+A232V+Q245R, T022A+N076D+S103A+A232V+Q245R, T022A+N076D+S103A+V104I+Q245R, T022A+N076D+V104I+Q245R, T022A+N076D+A232V+Q245R, T022A+N076D+V104I+A232V, T022A+N076D+S103A+Q245R, T022A+N076D+S103A+A232V, T022A+N076D+S103A+V104I, T022A+N076D+S103A, T022A+N076D+V104I, T022A+N076D+A232V, T022A+N076D+Q245R, G020R+S101A+T213A, G020R+R045T+S101A, G020R+S101A+G211Q, S024R+S101A+T213A, G020R+T022W+S101A, S024R+S101A+G211Q, N043R+S101A+T213A, N043R+R045T+S101A, S024R+N043R+S101A, S024R+R045T+S101A, S101A+G211Q+T213A, G020R+N043R+S101A, R045T+S101A+T213A, T022W+S024R+S101A, T022R+S101G+V104I, T022W+S101A+T213A, T022R+S101G+Q245R, T022W+S101A+G211Q, G020R+S024R+Q245R, T022R+S101G+V104I+Q245R, T022R+S101G+A232V, T022R+S101G+S103A, T022R+S101G+S103A+V104I, T022R+S101G+S103A+Q245R, T022R+S101G+A232V+Q245R, T022R+S101G+V104I+A232V, G020R+S024R+S101A+T213A, G020R+S024R+T213AS101N+V104I+A232V, S101N+S103A+V104I, S101N+S103A+A232V, S101N+S103A+V104I+A232V, G020R+S103A+A232V, G020R+V104I+A232V, G020R+S103A+V104I+A232V, G020R+S103A+V104I, N018R+N076D+N116A, N018R+N076D+T213A, N018R+N076D+S101A, N018R+N076D+A215F или N018R+N076D+G211Q, и где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
2. Выделенный вариант субтилизина по п. 1, где указанными положениями являются T022A+N076D+S103A+V104I+A232V+Q245R, и где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
3. Выделенный вариант субтилизина по п. 1, где указанные положения выбраны из T022R+S103A+V104I+A232V+Q245R, T022A+N076D+S103A+V104I+A232V, T022A+N076D+V104I+A232V+Q245R, T022A+N076D+S103A+A232V+Q245R или T022A+N076D+S103A+V104I+Q245R, и где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
4. Выделенный вариант субтилизина п. 1, где указанные положения выбраны из T022R+S103A+V104I+A232V, T022R+V104I+A232V+Q245R, T022R+S103A+V104I+Q245R, T022R+S103A+A232V+Q245R, T022A+N076D+V104I+Q245R, T022A+N076D+A232V+Q245R, T022A+N076D+V104I+A232V, T022A+N076D+S103A+Q245R, T022A+N076D+S103A+A232V, T022A+N076D+S103A+V104I, T022R+S101G+V104I+Q245R, T022R+S101G+S103A+V104I, T022R+S101G+S103A+Q245R, T022R+S101G+A232V+Q245R, T022R+S101G+V104I+A232V, G020R+S024R+S101A+T213A, S101N+S103A+V104I+A232V или G020R+S103A+V104I+A232V, где указанные положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
5. Выделенный вариант субтилизина п. 1, где указанные положения выбраны из T022R+V104I+Q245R, G020R+S024R+S101A, T022R+S103A+V104I, T022R+V104I+A232V, T022R+S103A+Q245R, T022R+A232V+Q245R, T022R+S103A+A232V, T022A+N076D+S103A, T022A+N076D+V104I, T022A+N076D+A232V, T022A+N076D+Q245R, G020R+S101A+T213A, G020R+R045T+S101A, G020R+S101A+G211Q, S024R+S101A+T213A, G020R+T022W+S101A, S024R+S101A+G211Q, N043R+S101A+T213A, N043R+R045T+S101A, S024R+N043R+S101A, S024R+R045T+S101A, S101A+G211Q+T213A, G020R+N043R+S101A, R045T+S101A+T213A, T022W+S024R+S101A, T022R+S101G+V104I, T022W+S101A+T213A, T022R+S101G+Q245R, T022W+S101A+G211Q, G020R+S024R+Q245R, T022R+S101G+A232V, T022R+S101G+S103A, G020R+S024R+T213A, S101N+V104I+A232V, S101N+S103A+V104I, S101N+S103A+A232V, G020R+S103A+A232V, G020R+V104I+A232V, G020R+S103A+V104I, N018R+N076D+N116A, N018R+N076D+T213A, N018R+N076D+S101A, N018R+N076D+A215F или N018R+N076D+G211Q, где указанные положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
6. Выделенный вариант субтилизина, где указанный вариант субтилизина представляет собой зрелую форму, обладающую протеолитической активностью, и содержит аминокислотную последовательность, включающую комбинацию замен аминокислот в положениях, выбранных из S103A+V104I+S188D, S188D+Q245R+N248D, S103A+S188D+Q245R, V104I+S188D+Q245R, S103A+V104I+S188D+Q245R, T022A+S103A+S188D, T022A+V104I+S188D, T022A+S103A+V104I+S188D, G159D+S188D+Q245R, S103A+S188D+A232V, S103A+G159D+S188D, V104I+S188D+A232V, V104I+G159D+S188D, S103A+V104I+S188D+A232V, S103A+V104I+G159D+S188D, S103A+S188D+N248D, V104I+S188D+N248D, S103A+V104I+S188D+N248D, G159D+S188D+N248D, S188D+A232V+Q245R, S103A+S188D+A232V+Q245R, V104I+S188D+A232V+Q245R, S103A+V104I+S188D+A232V+Q245R, T022A+S188D+Q245R, T022A+S103A+S188D+Q245R, T022A+V104I+S188D+Q245R, S103A+S188D+E271F, V104I+S188D+E271F, T022A+S103A+V104I+S188D+Q245R, S103A+V104I+S188D+E271F, T022A+S188D+N248D, T022A+S103A+S188D+N248D, T022A+V104I+S188D+N248D, S103A+S188D+Q245R+N248D, T022A+S103A+V104I+S188D+N248D, V104I+S188D+Q245R+N248D, S103A+V104I+S188D+Q245R+N248D, T022A+S188D+A232V, T022A+S103A+S188D+A232V, T022A+V104I+S188D+A232V, T022A+S103A+V104I+S188D+A232V, S188D+A232V+N248D, S188D+Q245R+E271F, T022A+S188D+E271F, T022A+S188D+Q245R+N248D, S103A+S188D+A232V+N248D, S103A+S188D+Q245R+E271F, S188D+N248D+E271F, V104I+S188D+A232V+N248D, V104I+S188D+Q245R+E271F, T022A+S103A+S188D+E271F, T022A+S103A+S188D+Q245R+N248D, S103A+S188D+N248D+E271F, T022A+V104I+S188D+E271F, T022A+V104I+S188D+Q245R+N248D, S103A+V104I+S188D+Q245R+E271F, S103A+V104I+S188D+A232V+N248D, V104I+S188D+N248D+E271F, S188D+A232V+E271F, T022A+S103A+V104I+S188D+E271F, T022A+S103A+V104I+S188D+Q245R+N248D, S103A+V104I+S188D+N248D+E271F, S103A+S188D+A232V+E271F, V104I+S188D+A232V+E271F, T022A+S188D+A232V+Q245R, T022A+S188D+Q245R+E271F, T022A+S188D+N248D+E271F, S103A+V104I+S188D+A232V+E271F, T022A+S103A+S188D+A232V+Q245R, S188D+A232V+Q245R+N248D, S188D+Q245R+N248D+E271F, T022A+S188D+A232V+N248D, T022A+S103A+S188D+Q245R+E271F, T022A+S103A+S188D+N248D+E271F, T022A+V104I+S188D+A232V+Q245R, S103A+S188D+A232V+Q245R+N248D, S103A+S188D+Q245R+N248D+E271F, T022A+S103A+S188D+A232V+N248D, T022A+V104I+S188D+N248D+E271F, T022A+V104I+S188D+Q245R+E271F, S188D+A232V+Q245R+E271F, S188D+A232V+N248D+E271F, T022A+S103A+V104I+S188D+A232V+Q245R, V104I+S188D+A232V+Q245R+N248D, T022A+V104I+S188D+A232V+N248D, V104I+S188D+Q245R+N248D+E271F, T022A+S188D+A232V+E271F, T022A+S103A+V104I+S188D+Q245R+E271F, T022A+S103A+V104I+S188D+N248D+E271F, T022A+S188D+Q245R+N248D+E271F, S103A+S188D+A232V+Q245R+E271F, S103A+S188D+A232V+N248D+E271F, S103A+V104I+S188D+Q245R+N248D+E271F, T022A+S103A+V104I+S188D+A232V+N248D, S103A+V104I+S188D+A232V+Q245R+N248D, T022A+S103A+S188D+A232V+E271F, V104I+S188D+A232V+N248D+E271F, V104I+S188D+A232V+Q245R+E271F, T022A+S103A+S188D+Q245R+N248D+E271F, T022A+V104I+S188D+A232V+E271F, T022A+S188D+A232V+N248D+E271F, T022A+S188D+A232V+Q245R+N248D, S188D+A232V+Q245R+N248D+E271F, T022A+V104I+S188D+Q245R+N248D+E271F, S103A+V104I+S188D+A232V+N248D+E271F, S103A+V104I+S188D+A232V+Q245R+E271F, T022A+S188D+A232V+Q245R+E271F, T022A+S103A+V104I+S188D+A232V+E271F, T022A+S103A+S188D+A232V+N248D+E271F, T022A+S103A+S188D+A232V+Q245R+N248D, S103A+S188D+A232V+Q245R+N248D+E271F, T022A+V104I+S188D+A232V+Q245R+N248D, T022A+S103A+S188D+A232V+Q245R+E271F, T022A+V104I+S188D+A232V+N248D+E271F, T022A+S188D+A232V+Q245R+N248D+E271F, V104I+S188D+A232V+Q245R+N248D+E271F, T022A+V104I+S188D+A232V+Q245R+E271F, G159D+S188D+Q245R+N248D, T022A+G159D+S188D, T022A+S103A+G159D+S188D, T022A+V104I+G159D+S188D, T022A+S103A+V104I+G159D+S188D, S103A+G159D+S188D+Q245R, V104I+G159D+S188D+Q245R, S103A+V104I+G159D+S188D+Q245R, S103A+G159D+S188D+N248D, V104I+G159D+S188D+N248D, S103A+V104I+G159D+S188D+N248D, G159D+S188D+A232V, S103A+G159D+S188D+A232V, V104I+G159D+S188D+A232V, S103A+V104I+G159D+S188D+A232V, T022A+G159D+S188D+Q245R, G159D+S188D+E271F, T022A+S103A+G159D+S188D+Q245R, S103A+G159D+S188D+E271F, T022A+V104I+G159D+S188D+Q245R, V104I+G159D+S188D+E271F, T022A+S103A+V104I+G159D+S188D+Q245R, S103A+V104I+G159D+S188D+E271F, T022A+G159D+S188D+N248D, T022A+S103A+G159D+S188D+N248D, T022A+V104I+G159D+S188D+N248D, S103A+G159D+S188D+Q245R+N248D, T022A+S103A+V104I+G159D+S188D+N248D, V104I+G159D+S188D+Q245R+N248D, S103A+V104I+G159D+S188D+Q245R+N248D, T022A+G159D+S188D+A232V, T022A+S103A+G159D+S188D+A232V, T022A+V104I+G159D+S188D+A232V, G159D+S188D+A232V+Q245R, T022A+S103A+V104I+G159D+S188D+A232V, S103A+G159D+S188D+A232V+Q245R, G159D+S188D+Q245R+E271F, V104I+G159D+S188D+A232V+Q245R, G159D+S188D+A232V+N248D, T022A+G159D+S188D+E271F, S103A+G159D+S188D+Q245R+E271F, G159D+S188D+N248D+E271F, S103A+V104I+G159D+S188D+A232V+Q245R, S103A+G159D+S188D+A232V+N248D, T022A+G159D+S188D+Q245R+N248D, T022A+S103A+G159D+S188D+E271F, V104I+G159D+S188D+Q245R+E271F, T022A+V104I+G159D+S188D+E271F, V104I+G159D+S188D+A232V+N248D, S103A+G159D+S188D+N248D+E271F, T022A+S103A+G159D+S188D+Q245R+N248D, S103A+V104I+G159D+S188D+Q245R+E271F, V104I+G159D+S188D+N248D+E271F, G159D+S188D+A232V+E271F, T022A+S103A+V104I+G159D+S188D+E271F, S103A+V104I+G159D+S188D+A232V+N248D, T022A+V104I+G159D+S188D+Q245R+N248D, S103A+V104I+G159D+S188D+N248D+E271F, S103A+G159D+S188D+A232V+E271F, V104I+G159D+S188D+A232V+E271F, T022A+G159D+S188D+N248D+E271F, T022A+G159D+S188D+A232V+Q245R, T022A+G159D+S188D+Q245R+E271F, S103A+V104I+G159D+S188D+A232V+E271F, G159D+S188D+Q245R+N248D+E271F, T022A+S103A+G159D+S188D+A232V+Q245R, T022A+S103A+G159D+S188D+Q245R+E271F, T022A+S103A+G159D+S188D+N248D+E271F, T022A+G159D+S188D+A232V+N248D, G159D+S188D+A232V+Q245R+N248D, T022A+V104I+G159D+S188D+Q245R+E271F, T022A+V104I+G159D+S188D+N248D+E271F, T022A+V104I+G159D+S188D+A232V+Q245R, S103A+G159D+S188D+Q245R+N248D+E271F, G159D+S188D+A232V+Q245R+E271F, G159D+S188D+A232V+N248D+E271F, T022A+S103A+G159D+S188D+A232V+N248D, V104I+G159D+S188D+Q245R+N248D+E271F, S103A+G159D+S188D+A232V+Q245R+N248D, T022A+G159D+S188D+A232V+E271F, T022A+G159D+S188D+Q245R+N248D+E271F, S103A+G159D+S188D+A232V+Q245R+E271F, T022A+V104I+G159D+S188D+A232V+N248D, V104I+G159D+S188D+A232V+Q245R+N248D, S103A+G159D+S188D+A232V+N248D+E271F, T022A+S103A+G159D+S188D+A232V+E271F, V104I+G159D+S188D+A232V+Q245R+E271F, V104I+G159D+S188D+A232V+N248D+E271F, T022A+V104I+G159D+S188D+A232V+E271F, G159D+S188D+A232V+Q245R+N248D+E271F, T022A+G159D+S188D+A232V+N248D+E271F, T022A+G159D+S188D+A232V+Q245R+E271F, T022A+G159D+S188D+A232V+Q245R+N248D, S101G+S103A+S188D, S101G+V104I+S188D, S101G+S103A+V104I+S188D, и где указанные положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
7. Выделенный вариант субтилизина по п. 6, где указанные положения выбраны из T022A+S103A+V104I+S188D+Q245R+N248D, T022A+S103A+V104I+S188D+A232V+Q245R, T022A+S103A+V104I+S188D+Q245R+E271F, T022A+S103A+V104I+S188D+N248D+E271F, S103A+V104I+S188D+Q245R+N248D+E271F, T022A+S103A+V104I+S188D+A232V+N248D, S103A+V104I+S188D+A232V+Q245R+N248D, T022A+S103A+S188D+Q245R+N248D+E271F, T022A+V104I+S188D+Q245R+N248D+E271F, S103A+V104I+S188D+A232V+N248D+E271F, S103A+V104I+S188D+A232V+Q245R+E271F, T022A+S103A+V104I+S188D+A232V+E271F, T022A+S103A+S188D+A232V+N248D+E271F, T022A+S103A+S188D+A232V+Q245R+N248D, S103A+S188D+A232V+Q245R+N248D+E271F, T022A+V104I+S188D+A232V+Q245R+N248D, T022A+S103A+S188D+A232V+Q245R+E271F, T022A+V104I+S188D+A232V+N248D+E271F, T022A+S188D+A232V+Q245R+N248D+E271F, V104I+S188D+A232V+Q245R+N248D+E271F, T022A+V104I+S188D+A232V+Q245R+E271F, T022A+S103A+V104I+G159D+S188D+Q245R, T022A+S103A+V104I+G159D+S188D+N248D, S103A+V104I+G159D+S188D+Q245R+N248D, T022A+S103A+V104I+G159D+S188D+A232V, S103A+V104I+G159D+S188D+A232V+Q245R, T022A+S103A+G159D+S188D+Q245R+N248D, S103A+V104I+G159D+S188D+Q245R+E271F, T022A+S103A+V104I+G159D+S188D+E271F, S103A+V104I+G159D+S188D+A232V+N248D, T022A+V104I+G159D+S188D+Q245R+N248D, S103A+V104I+G159D+S188D+N248D+E271F, S103A+V104I+G159D+S188D+A232V+E271F, T022A+S103A+G159D+S188D+A232V+Q245R, T022A+S103A+G159D+S188D+Q245R+E271F, T022A+S103A+G159D+S188D+N248D+E271F, T022A+V104I+G159D+S188D+Q245R+E271F, T022A+V104I+G159D+S188D+N248D+E271F, T022A+V104I+G159D+S188D+A232V+Q245R, S103A+G159D+S188D+Q245R+N248D+E271F, T022A+S103A+G159D+S188D+A232V+N248D, V104I+G159D+S188D+Q245R+N248D+E271F, S103A+G159D+S188D+A232V+Q245R+N248D, T022A+G159D+S188D+Q245R+N248D+E271F, S103A+G159D+S188D+A232V+Q245R+E271F, T022A+V104I+G159D+S188D+A232V+N248D, V104I+G159D+S188D+A232V+Q245R+N248D, S103A+G159D+S188D+A232V+N248D+E271F, T022A+S103A+G159D+S188D+A232V+E271F, V104I+G159D+S188D+A232V+Q245R+E271F, V104I+G159D+S188D+A232V+N248D+E271F, T022A+V104I+G159D+S188D+A232V+E271F, G159D+S188D+A232V+Q245R+N248D+E271F, T022A+G159D+S188D+A232V+N248D+E271F, T022A+G159D+S188D+A232V+Q245R+E271F или T022A+G159D+S188D+A232V+Q245R+N248D, и где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
8. Выделенный вариант субтилизина по п. 6, где указанные положения выбраны из S103A+V104I+S188D+A232V+Q245R, T022A+S103A+V104I+S188D+Q245R, T022A+S103A+V104I+S188D+N248D, S103A+V104I+S188D+Q245R+N248D, T022A+S103A+V104I+S188D+A232V, T022A+S103A+S188D+Q245R+N248D, T022A+V104I+S188D+Q245R+N248D, S103A+V104I+S188D+Q245R+E271F, S103A+V104I+S188D+A232V+N248D, T022A+S103A+V104I+S188D+E271F, S103A+V104I+S188D+N248D+E271F, S103A+V104I+S188D+A232V+E271F, T022A+S103A+S188D+A232V+Q245R, T022A+S103A+S188D+Q245R+E271F, T022A+S103A+S188D+N248D+E271F, T022A+V104I+S188D+A232V+Q245R, S103A+S188D+A232V+Q245R+N248D, S103A+S188D+Q245R+N248D+E271F, T022A+S103A+S188D+A232V+N248D, T022A+V104I+S188D+N248D+E271F, T022A+V104I+S188D+Q245R+E271F, V104I+S188D+A232V+Q245R+N248D, T022A+V104I+S188D+A232V+N248D, V104I+S188D+Q245R+N248D+E271F, T022A+S188D+Q245R+N248D+E271F, S103A+S188D+A232V+Q245R+E271F, S103A+S188D+A232V+N248D+E271F, T022A+S103A+S188D+A232V+E271F, V104I+S188D+A232V+N248D+E271F, V104I+S188D+A232V+Q245R+E271F, T022A+V104I+S188D+A232V+E271F, T022A+S188D+A232V+N248D+E271F, T022A+S188D+A232V+Q245R+N248D, S188D+A232V+Q245R+N248D+E271F, T022A+S188D+A232V+Q245R+E271F, T022A+S103A+V104I+G159D+S188D, S103A+V104I+G159D+S188D+Q245R, S103A+V104I+G159D+S188D+N248D, S103A+V104I+G159D+S188D+A232V, T022A+S103A+G159D+S188D+Q245R, T022A+V104I+G159D+S188D+Q245R, S103A+V104I+G159D+S188D+E271F, T022A+S103A+G159D+S188D+N248D, T022A+V104I+G159D+S188D+N248D, S103A+G159D+S188D+Q245R+N248D, V104I+G159D+S188D+Q245R+N248D, T022A+S103A+G159D+S188D+A232V, T022A+V104I+G159D+S188D+A232V, S103A+G159D+S188D+A232V+Q245R, V104I+G159D+S188D+A232V+Q245R, S103A+G159D+S188D+Q245R+E271F, S103A+G159D+S188D+A232V+N248D, T022A+G159D+S188D+Q245R+N248D, T022A+S103A+G159D+S188D+E271F, V104I+G159D+S188D+Q245R+E271F, T022A+V104I+G159D+S188D+E271F, V104I+G159D+S188D+A232V+N248D, S103A+G159D+S188D+N248D+E271F, V104I+G159D+S188D+N248D+E271F, S103A+G159D+S188D+A232V+E271F, V104I+G159D+S188D+A232V+E271F, T022A+G159D+S188D+N248D+E271F, T022A+G159D+S188D+A232V+Q245R, T022A+G159D+S188D+Q245R+E271F, G159D+S188D+Q245R+N248D+E271F, T022A+G159D+S188D+A232V+N248D, G159D+S188D+A232V+Q245R+N248D, G159D+S188D+A232V+Q245R+E271F, G159D+S188D+A232V+N248D+E271F или T022A+G159D+S188D+A232V+E271F, и где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
9. Выделенный вариант субтилизина по п. 6, где указанные положения выбраны из S103A+V104I+S188D+Q245R, T022A+S103A+V104I+S188D, S103A+V104I+S188D+A232V, S103A+V104I+G159D+S188D, S103A+V104I+S188D+N248D, S103A+S188D+A232V+Q245R, V104I+S188D+A232V+Q245R, T022A+S103A+S188D+Q245R, T022A+V104I+S188D+Q245R, S103A+V104I+S188D+E271F, T022A+S103A+S188D+N248D, T022A+V104I+S188D+N248D, S103A+S188D+Q245R+N248D, V104I+S188D+Q245R+N248D, T022A+S103A+S188D+A232V, T022A+V104I+S188D+A232V, T022A+S188D+Q245R+N248D, S103A+S188D+A232V+N248D, S103A+S188D+Q245R+E271F, V104I+S188D+A232V+N248D, V104I+S188D+Q245R+E271F, T022A+S103A+S188D+E271F, S103A+S188D+N248D+E271F, T022A+V104I+S188D+E271F, V104I+S188D+N248D+E271F, S103A+S188D+A232V+E271F, V104I+S188D+A232V+E271F, T022A+S188D+A232V+Q245R, T022A+S188D+Q245R+E271F, T022A+S188D+N248D+E271F, S188D+A232V+Q245R+N248D, S188D+Q245R+N248D+E271F, T022A+S188D+A232V+N248D, S188D+A232V+Q245R+E271F, S188D+A232V+N248D+E271F, T022A+S188D+A232V+E271F, G159D+S188D+Q245R+N248D, T022A+S103A+G159D+S188D, T022A+V104I+G159D+S188D, S103A+G159D+S188D+Q245R, V104I+G159D+S188D+Q245R, S103A+G159D+S188D+N248D, V104I+G159D+S188D+N248D, S103A+G159D+S188D+A232V, V104I+G159D+S188D+A232V, T022A+G159D+S188D+Q245R, S103A+G159D+S188D+E271F, V104I+G159D+S188D+E271F, T022A+G159D+S188D+N248D, T022A+G159D+S188D+A232V, G159D+S188D+A232V+Q245R, G159D+S188D+Q245R+E271F, G159D+S188D+A232V+N248D, T022A+G159D+S188D+E271F, G159D+S188D+N248D+E271F, G159D+S188D+A232V+E271F или S101G+S103A+V104I+S188D, и где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
10. Выделенный вариант субтилизина по п. 6, где указанные положения выбраны из S103A+V104I+S188D, S188D+Q245R+N248D, S103A+S188D+Q245R, V104I+S188D+Q245R, T022A+S103A+S188D, T022A+V104I+S188D, G159D+S188D+Q245R, S103A+S188D+A232V, S103A+G159D+S188D, V104I+S188D+A232V, V104I+G159D+S188D, S103A+S188D+N248D, V104I+S188D+N248D, G159D+S188D+N248D, S188D+A232V+Q245R, T022A+S188D+Q245R, S103A+S188D+E271F, V104I+S188D+E271F, T022A+S188D+N248D, T022A+S188D+A232V, S188D+A232V+N248D, S188D+Q245R+E271F, T022A+S188D+E271F, S188D+N248D+E271F, S188D+A232V+E271F, T022A+G159D+S188D, G159D+S188D+A232V, G159D+S188D+E271F, S101G+S103A+S188D или S101G+V104I+S188D, и где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
11. Выделенный вариант субтилизина, где указанный вариант субтилизина представляет собой зрелую форму, обладающую протеолитической активностью, и содержит аминокислотную последовательность, включающую комбинацию замен аминокислот, выбранную из S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S101G-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S101G-S103A-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S101G-S103A-V104I-S188D-A232V-Q245R-N248D-E271F, T022A-S101G-S103A-V104I-G159D-S188D-Q245R-N248D-E271F, T022A-S101G-S103A-V104I-G159D-S188D-A232V-N248D-E271F, T022A-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-E271F, T022A-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248D, S101G-S103A-V104I-N116L-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S103A-V104I-N116L-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S101G-V104I-N116L-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S101G-S103A-N116L-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S101G-S103A-V104I-N116L-S188D-A232V-Q245R-N248D-E271F, T022A-S101G-S103A-V104I-N116L-G159D-A232V-Q245R-N248D-E271F, T022A-S101G-S103A-V104I-N116L-G159D-S188D-Q245R-N248D-E271F, T022A-S101G-S103A-V104I-N116L-G159D-S188D-A232V-N248D-E271F, T022A-S101G-S103A-V104I-N116L-G159D-S188D-A232V-Q245R -E271F, T022A-S101G-S103A-V104I-N116L-G159D-S188D-A232V-Q245R-N248D-E271F, G020R-N062E-S078G-G118S-S188D-N248D-H249R, S024R-N062E-G118R-A158E-S188D, T022A-S024R-T033S-G118R-S166D-S188D, S024R-N062E-S078G-G118S-S188D-Q245R-N248D, G020R-S024R-N062E-S078D-G118S-P129E-G159D, G020R-S024R-N062E-S078G-G118D-P129E, G020R-S024R-N062E-S078G-G118S-P129E-G159D-S188D-Q245R-N248D, G020R-N062E-S078G-G118S-G159D-Q245R-N248D, S024R-N116L-A158E-S166D, G020R-N062E-S078G-G118S-G159D-S188D-H249R, G020R-S024R-S078G-G118D-P129E-G159D-S188D, S024R-G118R-S166D, T022A-S024R-T033S-G118R-A158E-S166D-A273V, G020R-N062E-S078G-G118S-P129E-G159D-S188D-Q245R, T022A-G118R-A158E-S166D, G020R-S024R-N062E-S078D-G118S-G159D-S188D-Q245R-N248D, T022A-S024R-G118R-S166D-S188D, N062E-S078G-G118S-G159D, T022A-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248E-E271H, G020R-S024R-N062E-S078G-G118D-Q245R-N248D, S024R-T033S-N116L-A158E-S166D, S024R-N062E-S078G-G118D-P129E-G159D-Q245R, G020R-N062E-S078D-G118S-Q245R, T022A-S024R-N062E-N116L-A158E, G020R-N062E-S078G-G118D-P129E, S024R-T033S-N062E-N116L-G118R-S188D, G020R-N062E-S078G-G118D-P129E-G159D-S188D-H249R, S024R-S078G-G118S-P129E-G159D-Q245R-N248D, G020R-N062E-S078G-G118D-S188D-H249R, T022Q-S101G-S103A-V104I-G159D-S188D-A232L-Q245T-N248D-E271T, S078G-G118S-P129E-G159D-S188D-Q245R-N248D-H249R, T022Q-S101G-S103A-V104I-G159D-S188E-A232V-Q245T-N248H-E271T, G020R-S024R-S078D-G118S-G159D-S188D-H249R, G020R-S078D-G118S-P129E-G159D-Q245R-N248D-H249R, S024R-N062E-N116L-G118R, T022A-S024R-N116L-G118R-A158E-S188D, S024R-N062E-S078G-G118S-P129E-Q245R, S024R-N062E-G118R-A158E, G020R-N062E-S078G-G118S-G159D-S188D, G020R-S024R-S078D-G118S-G159D-S188D-Q245R-N248D-H249R, S024R-S078D-G118S-P129E-G159D-Q245R, A098Q-S099T-G102A-S103G, T022A-G118R-S166D-S188D, S024R-S078D-G118S-G159D-S188D-H249R, T022A-S024R-N116L-G118R-S166D-S188D, T022A-N062E-G118R-A158E, G020R-S078G-G118D-G159D-S188D-Q245R, S024R-N062E-S078G-G118D-G159D-S188D-Q245R, T022Q-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248E-E271F, T022A-S101G-S103A-V104I-G159D-S188E-A232S-Q245R-N248E-E271F, G020R-N062E-S078G-G118S-P129E-N248D-H249R, S024R-N116L-G118R-S128I-S166D-S188D, G020R-S024R-N062E-S078G-G118D-P129E-G159D-S188D-H249R, G020R-S024R-N062E-S078G-G118D-G159D-S188D-N248D, G020R-S024R-S078G-G118D-P129E-S188D-Q245R-N248D, S024R-N062E-S078D-G118S-S188D-H249R, T033S-N062E-G118R, T022Q-S101G-S103A-V104I-G159D-S188D-A232S-Q245T-N248E-E271H, T033S-G118R-A158E-S166D, T022Q-S101G-S103A-V104I-G159D-S188E-A232S-Q245V-N248H, G020R-S024R-S078D-G118D-S188D-Q245R-N248D-H249R, T022A-S024R-N062E, G020R-S024R-N062E-S078D-G118D-G159D-S188D-R247L, T022Q-S101G-S103A-V104I-G159D-S188D-A232S-Q245R-N248E-E271T, G020R-N062E-S078G-G118S-P129E-G159D-Q245R-N248D, T022Q-S101G-S103A-V104I-G159D-S188E-A232S-Q245R-N248H, P014S-G020R-S024R-S078G-G118S-P129E-S188D-Q245R-N248D, T022A-S101G-S103A-V104I-G159D-S188E-A232V-Q245R-N248E-E271L, G020R-S078D-G118D-G159D-S188D-H249R, T022Q-S101G-S103A-V104I-G159D-S188E-A232L-Q245R-N248D-E271T, T022A-S101G-S103A-V104I-G159D-S188E-A232T-Q245T-N248E-E271L, S024R-N062E-S078D-G118S-S188D-Q245R, T022Q-S101G-S103A-V104I-G159D-S188E-A232L-Q245R-N248E-E271H, G020R-S078D-G118S-P129E-G159D-H249R, T022A-S101G-S103A-V104I-G159D-S188D-A232S-Q245R-N248H-E271F, T022A-S101G-S103A-V104I-G159D-S188D-A232L-Q245R-N248H-E271H, S024F-T033S-S101G-S103A-V104I-N116A-Y209A-T213A-A232V, S024F-S101G-S103A-V104I-N116L-S128N-Y209V-G211Q-T213A-A232V, S024R-S078G-G118S-G159D-S188D-Q245R-N248D, G020R-S024R-S078G-G118S-S188D-Q245R, G020R-S024R-N062E-S078D-G118D-P129E-G159D-Q245R, S024R-S078D-G118D-G159D-S188D-H249R, G020R-S078G-G118S-G159D-S188D-H249R, G020R-S024R-N062E-S078G-G118S-S188D-N248D, S078D-G118S-G159D-Q245R, G020R-S024R-N062E-S078G-G118D-P129E-G159D-N248D-H249R, S024R-N062E-S078G-G118S-P129E-Q245R-N248D, G020R-S078G-G118S-P129E-S188D-Q245R-N248D, S024R-N062E-S078G-G118S-G159D-Q245R, G020R-N062E-S078D-G118D-P129E-Q245R, T022A-S101G-S103A-V104I-G159D-S188E-P210L-A232T-Q245T-N248D-E271T, G020R-S024R-N062E-S078G-G118S-P129E, G020R-S024R-N062E-S078G-G118D-G159D-S188D-H249R, T022Q-S101G-S103A-V104I-G159D-S188D-A232T-Q245V-N248D-E271L, A016S-T022A-S101G-S103A-V104I-N116L-S128N-Y209V-A232V, T022A-S101G-S103A-V104I-G159D-S188E-A232S-Q245T-N248H-E271H, G020R-N062E-S078G-G118D-P129E-G159D-H249R, T022A-S101G-S103A-V104I-G159D-S188D-A232L-Q245R-N248E, G020R-S024R-S078G-G118D-G159D-S188D-Q245R, S024R-N062E-S078G-G118S-G159D-Q245R-N248D, G020R-S078G-G118S-G159D-S188D-Q245R, S024R-N062E-S078G-G118S-G159D-S188D, T022A-S101G-S103A-V104I-G159D-S188E-A232V-Q245R-N248H-E271H, G020R-S024R-N062E-S078G-G118D-G159D, T022A-S101G-S103A-V104I-G159D-S188D-A232T-V244A-Q245R-N248H-E271H, S024R-S078G-G118S-P129E-G159D-S188D-Q245R, T022Q-S101G-S103A-V104I-G159D-S188E-A232V-Q245V-N248D-E271F, G020R-S024R-N062E-S078G-G118D-G159D-S188D-Q245R, T022A-S024R-N062E-S128I-A158E, G020R-S024R-S078G-G118D-N248D-H249R, S024R-S078G-G118D-P129E-Q245R-H249R, T022A-S101G-S103A-V104I-G159D-S188D-A232T-Q236K-Q245T-N248E, N062E-S078G-G118S-G159D-Q245R-N248D, T033S-G118R-S166D, G020R-S024R-S078G-G118D-P129E-S188D-N248D, G020R-S078G-G118S-P129E-G159D-S188D-Q245R-N248D, A016S-T022A-S101G-S103A-V104I-L111V, T022Q-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248E-E271H, S024R-N116L-G118R-S128I-A158E-S188D-V199I, G020R-S024R-S078D-G118D-G159D-H249R, T022A-S101G-S103A-V104I-G159D-S188E-A232S-Q245R-N248E-E271T, G020R-S024R-S078D-G118S-S188D-N248D-H249R, T022A-S101G-S103A-V104I-G159D-S188D-A232T-Q245R-N248D-E271T, G020R-S024R-S078G-G118D-P129E-S188D-H249R, S024R-N062E-S078G-G118S-S188D-N248D, T022A-S101G-S103A-V104I-N116L-S128N-Y209A-G211Q-T213A-A232V, T022Q-S101G-S103A-V104I-G159D-S188D-A232T-Q245R-N248D-E271L, S024R-A048V-S078G-G118D-G159D-S188D-H249R, G020R-S024R-N062E-S078D-G118D, G102A-S103G-V104I, G020R-S024R-S078D-G118D-G159D-Q245R-N248D-H249R, T022A-T033S-G118R-A158E, T022A-S024R-N116L-S128I-S188D, G020R-S024R-S078D-G118D-P129E-G159D-Q245R-N248D-H249R, T022Q-S101G-S103A-V104I-G159D-S188E-A232T-Q245T-N248E-E271F, S024R-N062E-S078G-G118S-S188D-Q245R, S024R-N062E-S078G-G118D-G159D, G020R-S024R-N062E-S078D-G118S-P129E-G159D-Q245R, T022Q-S101G-S103A-V104I-G159D-S188E-A232S-Q245V-N248E-E271T, G020R-S078G-G118D-S188D-N248D-H249R, T022A-S024R-N062E-N116L-G118R-A158E-S166D-S188D, S078G-G118S-G159D-S188D-H249R, T022Q-S101G-S103A-V104I-G159D-S188E-A232T-Q245V-N248H-E271F, G020R-S078G-G118S-P129E-G159D-S240F-N248D-H249R, S024F-T033S-S101G-S103A-V104I-N116A-Y209A-G211Q-T213A-A232V, N062E-S078D-G118S-S188D-H249R, T022A-S101G-S103A-V104I-G159D-S188E-A232V-Q245V-N248D-E271T, G020R-S078G-G118S-G159D-S188D-Q245R-N248D, T022Q-S101G-S103A-V104I-G159D-S188D-A232T-Q245R-N248D-E271F, S024R-S078G-G118D-G159D-S188D-Q245R-N248D-H249R, G020R-S024R-N062E-S078D-G118D-Q245R, S024R-S078G-G118D-P129E-S188D-Q245R, N062E-S078G-G118S-S188D-Q245R-N248D-H249R, S024R-N062E-S078G-G118D-G159D-H249R, G020R-S078G-G118S-P129E-G159D-S188D-H249R, S024R-N062E-S078G-G118S-P129E-S188D-H249R, T022Q-S101G-S103A-V104I-G159D-S188E-A232L-Q245R-N248H, T022Q-S101G-S103A-V104I-G159D-S188D-A232V-Q245V-N248H-E271L, G020R-S101G-S103A-V104I-N116A-S128N-Y209V-G211Q-T213A-A232V, S024R-N062E-S078D-G118S-P129E-G159D-H249R, T022A-S101G-S103A-V104I-G159D-S188E-A232V-Q245R-N248D-E271H, G020R-N062E-S078G-G118D-P129E-S188D-H249R, T022Q-S101G-S103A-V104I-G159D-S188E-A232L-Q245V-N248E-E271L, G020K-T022L-S078G-G118S-P129E-G159D, S078G-G118S-S188D-Q245R-N248D, T022A-S024F-S101G-S103A-V104I-N116A-S128N-Y209A-A232V, G020R-S024R-N062E-S078G-G118S-G159D-N248D, N062E-G118R-S166D-S188D, T022Q-S101G-S103A-V104I-G159D-S188E-A232T-Q245R-N248E-E271T, T022A-S101G-S103A-V104I-G159D-S188D-A232S-Q245T-N248H-E271F, G020R-N062E-S078G-G118S-G159D-H249R, S024R-N062E-N116L-A158E, S024R-S078G-G118D-P129E-Q245R-N248D-H249R, G020R-S024R-N062E-S078D-G118S-P129E-S188D-Q245R, S024R-S078G-G118D-P129E-G159D-S188D-H249R, T022Q-S101G-S103A-V104I-G159D-S188E-A232L-Q245R-N248D-E271H, S024R-S078D-G118D-P129E-S188D-Q245R-N248D-H249R, G020R-S024R-S078G-G118D-P129E-G159D-N248D-H249R, T022A-S101G-S103A-V104I-G159D-S188E-A232V-Q245T-N248E-E271F, T022Q-S101G-S103A-V104I-G159D-S188D-A232L-Q245T-N248H-E271T, S024R-S078D-G118S-P129E-G159D, G020R-N062E-S078G-G118S-P129E-S188D-Q245R, T022A-S024R-N116L-G118R-S128I-S188D, T022A-S101G-S103A-V104I-G159D-S188E-A232L-Q245T-N248E-E271H, G020R-S024R-S078D-G118S-P129E-G159D-S188D-Q245R-N248D, G020R-S024R-N062E-S078G-G118S-P129E-S188D-N248D-H249R, T022A-S101G-S103A-V104I-G159D-S188E-A232V-Q245V-N248H-E271H, G020R-S078G-G118D-P129E-G159D-S188D-H249R, G020R-S024R-S078D-G118S-S188D-Q245R-H249R, T022A-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248E-E271F, T022A-S101G-S103A-V104I-G159D-S188D-A232T-Q245R-N248D-E271F, S024R-N062E-S078G-G118S-S188D-N248D-H249R, G020R-N062E-S078G-G118S-G159D-Q245R, S024R-N062E-G118R-A158E-S166D-S188D, S024R-N062E-S078G-G118S-P129E-G159D-N248D-H249R, G020R-S024R-N062E-S078G-G118S-P129E-A158T-G159D-Q245R-N248D, T022Q-S101G-S103A-V104I-G159D-S188E-A232T-Q245R-N248E-E271L, G020R-S024R-N062E-S078D-G118S-S188D-H249R, S024R-S078G-G118S-S188D-N248D, T022A-S101G-S103A-V104I-G159D-S188E-A232L-Q245V-N248E-E271H, T022Q-S101G-S103A-V104I-G159D-S188D-A232L-Q245T-N248H-E271H, S099G-S101G, T022Q-S101G-S103A-V104I-G159D-S188D-A232T-Q245V-N248D-E271F, G020R-N062E-S078D-G118S-P129E-G159D-Q245R, T022A-S101G-S103A-V104I-G159D-S188E-A232T-Q245T-N248H-E271H, T022Q-S101G-S103A-V104I-G159D-S188E-A232V-Q245V-N248H-E271L, T022A-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248E-E271T, T022Q-S101G-S103A-V104I-G159D-S188E-A232L-Q245T-N248E-E271T, T022Q-S101G-S103A-V104I-G159D-S188E-A232L-Q245V-N248E-E271H, T022Q-S101G-S103A-V104I-G159D-S188D-A232T-Q245R-N248H-E271L, G020R-S078G-G118S-P129E-G159D-S188D-Q245R-N248D-H249R, G020R-S024R-N062E-S078G-G118D-S188D-N248D, S024R-N062E-S078G-G118S-P129E-G159D, A016S-T022A-S101G-S103A-V104I-N116A-S128N-Y209V-G211Q-A232V, S024R-S078G-G118D-S188D-Q245R-N248D-H249R, G020R-S024R-S078D-G118D-G159D-S188D-Q245R-N248D-H249R, A016S-T022A-S101G-S103A-V104I-N116A-Y209V-G211Q-T213A-A232V, S024R-N062E-S078G-G118D-S188D-H249R, T022L-S078N-G118R-S166D-T213A-L217E, T022A-S101G-S103A-V104I-G159D-S188D-A232S-Q245R-N248E-E271H, S024R-S078G-G118D-G159D-N248D-H249R, S024R-S078G-G118D-P129E-S188D-Q245R-N248D, T022Q-S101G-S103A-V104I-G159D-S188E-A232T-Q245T-N248H-E271T, T022A-S101G-S103A-V104I-G159D-S188D-A232S-Q245R-N248H-E271H, G020R-S024R-S078G-G118S-G159D-S188D-Q245R-N248D, T022A-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271L, 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T022A-G118R-S166D, G020R-S024R-S078G-G118S-G159D-S188D-Q245R, T022A-S024R-T033S-N116L-G118R-A158E-S188D, T022Q-S101G-S103A-V104I-G159D-S188E-A232T-Q245V-N248D-E271F, G020R-S024R-S078D-G118S-P129E-Q245R-N248D-H249R, A016S-T022A-V104I-S128N, S024R-N062E-N076D-S101A-S103A-V104I-S188D-P210I-A232V-Q245R, T022L-G118R-S166D-T213A-L217E, S078D-G118S-P129E-G159D-S188D-Q245R-H249R, G195W-N269E, G020R-S024R-S078G-G118S-P129E-Q245V-N248E, N269W, T022A-T033S-G118R-A158E-S166D-S188D, G020R-N062E-S078G-G118D-P129E-G159D-Q245R-H249R, A016S-S101A-S103A, G097P-S099G-S103A-V104I, S024R-S078D-G118D-P129E-Q245R, T022Q-S101G-S103A-V104I-G159D-S188D-A232L-Q245R-N248E, S078G-G118D-P129E-S188D-Q245R, T033S-N062E-G118R-A158E, G020R-S078D-G118S-G159D-N248D, A016S-T022A-S103G-V104I-S128N-L148I, A016S-S101G-S103A-V104I-N116A-Y209A-A232V, S101A-S103A-V104I-S188D-M222S-A232V-Q245R, T022A-S024F-S101G-S103A-V104I-N116A-S128N-Y209V-T213A-A232V, A016S-T022A-S024F-S101G-S103A-V104I-N116A-Y209A-T213A-A232V, T022A-S101G-S103A-V104I-G159D-S188D-T213A-A232V-Q245R-N248D-E271F, A016S-S101G-S103A-V104I-N116L-Y209A-T213A-A232V, G020R-S078D-G118S-S188D, G020R-S078D-G118S-P129E-Q245R-N248D, T022L-S166D-T213A, G020R-S024R-S078D-G118S-P129E-G159D-S188D-N248D, S240G, S078G-G118S-P129E-Q245R-N248D, S024R-S078D-G118D-P129E-G159D-Q245R-N248D, S024R-S078G-G118S-G159D, T022A-S024R-N076D-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F-A272V, N062E-S078G-G118S-P129E-S188D-Q245R-N248D-H249R, S024F-S078N-G118R-S166D, S078G-G118S-P129E-N248D, N155W, A016S-S024F-S101G-S103A-V104I-N116A-S128N-Y209V-G211Q-T213A-A232V, G020R-N062E-S078D-G118D-P129E-Q245R-N248D, G020R-S078D-G118D-P129E-S188D-Q236R-N248D-H249R, T022A-S024R-N116L-A158E-S188D, S101G-S103A-V104I-N116A-S128N-Y209V-G211Q-T213A-A232V, S078D-G118S-S188D-Q245R-N248D, T022L-S024F-S078N-L217E, S099A-S101A-G102A, G020R-S078D-G118D-G159D-N248D-H249R, T022A-N076D-S101G-S103A-V104I-I107V-P129E-G159D-S188D-A232V-Q245R-N248D-E271F, T033S-N116L-G118R-S166D-S188D, S024R-G118R-S128I-A158E-S166D-S188D, G020R-S024R-N062E-S078G-G118S-P129E-G159D-Q245R-N248D-H249R, N062E-S078G-G118D-N248D-H249R, N062E-S078R-P129E-S188D-Q245R-N248D, S024R-N062E-G159D-S188D-Q245R, G020R-N062E-G118R-S188D-N248D-H249R, N062E-S078R-P129E-G159D, G020R-S024R-N062E-P129E-S188D-N248D, N062E-S078R-G118R-G159D-S188D-P210L-N248D, N062E-S078R-Q245R-N248D, G020R-G159D-S188D-H249R, G020R-G118R-G159D-S188D-N248D, G118R-P129E-G159D-S188D-H249R, N062E-S078R-G118R-P129E-N248D, N062E-S078R-G159D, N062E-G118R-P129E-S188D-Q245R, G020R-S024R-P129E-G159D-S188D-N248D, S024R-N062E-S078R-G159D-N248D, G020R-P129E-S188D-H249R, G020R-P129E-G159D-S188D-Q245R, A016S-S024F-S101G-S103A-V104I-N116A-S128N-S188D-Y209V-T213A-A232V, G020R-G159D-S188D-Q245R-N248D, G118R-P129E-G159D-Q245R-N248D, G020R-N062E-S078R-P129E-S188D-N248D, G020R-P129E-S188D-Q245R, S024R-N062E-S188D-N248D, S078R-L111I-N185I-I198L-L217E, G020R-P129E-S188D-N248D-H249R, G020R-N062E-G118R-G159D-N248D, G020K-T022L-G118R-P129E-S188D, G020R-G118R-P129E-S188D, G020R-P129E-G159D-Q245R-N248D, G020R-S024R-P129E-G159D, G020R-P129E-G159D-Q245R-N248D-H249R, S024R-P129E-G159D-S188D-Q245R-N248D-H249R, G020R-P129E-Q245R-N248D, N062E-S078R-G159D-N248D, S024R-G118R-P129E-G159D-S188D-Q245R-N248D-H249R, G020R-G061W-P129E-G159D-S188D-Q245R, T022A-S101G-S103A-V104I-G159D-S188D-M222S-A232V-Q245R-N248D-A270V-E271F, N062E-G159D-S188D-H249R, A016S-S024F-S101G-S103A-V104I-N116A-S128N-S130D-Y209V-T213A-A232V, G020R-S024R-P129E-G159D-N248D-H249R, P129E-G159D-S188D-H249R, G118R-P129E-G159D-S188D-K237R-N248D-H249R, G020R-G159D-S188D-Q245R, G020R-S188D-Q245R-N248D-H249R, T022A-S101G-S103A-V104I-P129S-L217Q-A232V-Q245R, G020R-S078R-G118R-P129E-S188D-N248D, G020R-S024R-G118R-G159D-S188D-Q245R-N248D, G020R-G118R-P129E-G159D-S188D, G020R-G159D-S188D, G020R-G118R-P129E-S188D-N248D, A016S-S024R-S101G-S103A-V104I-N116A-S128N-Y209V-T213A-A232V, G020K-N062E-S188D-T213A-M222S, A016S-S024F-S101D-S103A-V104I-N116A-S128N-Y209V-T213A-A232V, S024G-S078N-S101N-Q109G-S128A-L217Q, T022A-G097S-S101N-S103A-V104I-S128A-G159D-S188D-L217Q-M222S-A232V-Q245R-N248D-E271F, T022A-S101G-S103A-V104I-G159D-S188D-M222S-A232V-Q245R-N248D-E271F, G020R-P129E-Q245R, S024R-P129E-G159D-Q245R-N248D, S024R-G118R-S188D-N248D, G020R-P129E-H249R, S024R-P129E-Q245R-N248D, T022A-S101G-S103A-V104I-L217Q-A232V-Q245R, G020R-S078R-P129E-G159D-S188D-H249R, T022A-S101G-S103A-V104I-S128A-P129S-L217Q-A232V-Q245R, G118R-P129E-H249R, G020R-N062E-G118R, G020R-G118R-P129E-G159D-Q245R-N248D, T022A-S101Q-S103A-V104I-P129S-L217Q-A232V-Q245R, S024R-S078R-G118R-P129E-G159D-S188D, P129E-Q245R-N248D-H249R, S024G-G061S-S078N-S101N-Q109G-S128A-L217Q-N243V, S078R-V084A-P129E-Q245R, S024R-G118R-G159D-S188D-N248D, G020R-G118R-P129E-G159D-H249R, S078R-P129E-G159D, G061P-S078N-G097A-S101N-S128A-V203Y-L217Q, P086S-S087G-A088V-G097A-N117S-S128A-L217Q, S024R-S078R-G159D-S188D-Q245R-N248D-H249R, S078R-G118R-P129E, S024G-S078N-S101N-S128A-L217Q-N243V, S024R-S078R-G159D-S188D, G020R-G118R-P129E, G020R-S024R-P129E-G159D-S188D-H249R, S024R-G118R-G159D-S188D, S024R-G118R-P129E-G159D-S188D-Q245R-N248D, S024G-S078N-S101N-S128A-L217Q, S024R-G159D-S188D-Q245R, S024G-S078N-S101N-Q109G-L217Q-N243V-N248A, G020R-S078R-G159D-S188D-Q245R-N248D, S078R-G118R-P129E-S188D-N248D, S024R-S188D-Q245R-N248D, G020R-S024R-S101A-P210I-G211Q, G020R-G159D-N248D-H249R, G020R-P129E, G020K-N062E-N116L-T213A-M222S, S024G-S078N-A088T-S101N-Q109G-N116T-S128A-A158S-L217Q-N218S-L257G, A016S-S024F-S101G-S103A-V104I-N116A-S128N-S166D-Y209V-T213A-A232V, S024R-N062E-S078R-P129E-H249R, N018K-G020R-S024R-R045T-S101A-P210I-G211Q-T213A, G020R-S024R-G118R-P129E-G159D-S188D-N248D-H249R, G020R-S024R-P129E-H249R, G020R-G118R-S188D-Q245R-N248D-H249R, G020R-S024R-N062E-S078G-G118S-Q245R, G020R-S024R-G118R-P129E-G159D-N248D, S024G-S078N-S101N-Q109G-S128A-L217Q-S256R, S024G-S078N-S101N-S128A-A158S-L217Q-L257G, G020R-S024R-G118R-S188D-N248D, G020R-G118R-G159D-S188D-Q245R-N248D-H249R, G159D-S188D-Q245R-N248D, S078N-G118R-S166D-L217E-M222S, G020R-G118R-P129E-G159D-Q245R-N248D-H249R, T022A-S101Q-S103A-V104I-L124V-S128A-P129S-A232V-Q245R, S024R-N062E-Q245R, S188D-Q245R-N248D, S024R-G118R-P129E-S188D-H249R, T022A-G097A-S101Q-S103A-V104I-P129S-A232V-Q245R, G020R-G118R-S188D-Q245R-N248D, S024G-S078N-S101N-Q109G-N116T-S128A-L217Q-N243V-S256R, G020R-G118R, G020R-S024R-S188D, N062E-P129E-G159D-S188D-Q245R, G020R-G159D-Q245R-N248D, G020R-N043R-S101A-P210I-G211Q, G097A-S101N-S128A-L217Q, G020R-G118R-S188D-N248D, S024R-S188D-N248D, G020R-S024R-S188D-Q245R-N248D, G020R-S024R-G118R-P129E-G159D-N248D-H249R, G020R-P129E-N248D, G159D-S188D, P129E-G159D, G020R-S188D-H249R, P129E-G159D-Q245R-N248D, G020R-G118R-G159D-N248D-H249R, G118R-S188D-N248D-H249R, G020K-S101D-G102A-L217E-S240R, N043R-G102A-L217E, N062E-S078N-S101D-S240R, G102A-L217E, G020K-S128L-L217E-S240R, G020K-N062E-S078N-L217E-S240R, N043R-S101D-S128L-L217E-S240R, G020K-S078N-S101D-G102A-P210L, N043R-S101D-L217E, G020K-G100S-L217E-S240R, S078N-G100S, G020K-N043R-S101D-G102A-L217E, G100S-S240R, G020K-G102A-L217E, G020K-S024G-N062E-S078N-S101N-Q109N-N116L-S128A-S188D, G020K-L217E-S240R, N043R-S101D, N043R-S078N-S101D-S128L-S240R, N043R-S078N-S101D-L217E-S240R, G020K-S078N-S101D, N043R-S128L-S240R, G020K-N062E-N116L-N123G-T213A, G020K-N062E-G097S-S101G-Q109G-N116L-S128A, G020K-N062E-S078N-G097A-S101N-Q109N-N116L-S188D, N043R-S078N-S101D-G102A-L217E, G020K-S078N-S099G-S128L-L217E, G020K-N062E-S101D, G020K-S128L-L217E, G020K-S024G-N062E-S078N-G097S-S101G-Q109G-N116L-L217Q, N043R-G097A-S101D-S128L, G020K-S024G-N062E-S101Q-Q109N-N116L-S128A, G020K-S024G-N062E-S078N-S101N-N116L-S128A-L217Q, S078N-G102A-S240R, N043R-S078N-G100S-L217E-S240R, G020K-S078N-S128L-L217E-S240R, G020K-N062E-N116L-S128Q, G020K-S101G-L217E-S240R, G020K-S024G-N062E-S101N-Q109G-N116L-S128A-L217Q, G020K-N062E-S078N-G097S-S101N-N116L-L217Q, G020K-N062E-G097S-S101N-Q109G-N116L-L217Q, G020K-S024G-N062E-S078N-G097A-S101Q-Q109N-N116L-M222S, G020K-S024G-N062E-S078N-G097S-S101Q-Q109N-N116L-S128A-S188D-L217Q, G020K-N062E-S078N-S101N-Q109G-N116L-S188D-L217Q или G020K-N062E-S105G-N116L-S128L-T213A, где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
12. Выделенный вариант субтилизина, где указанный вариант субтилизина представляет собой зрелую форму, обладающую протеолитической активностью, и содержит аминокислотную последовательность, включающую комбинацию замен аминокислот, выбранную из T022A-S101G-S103A-V104I-N116L-G159D-S188D-A232V-Q245R-N248D, S024R-N116L-A158E-S188D, G020K-S024F-S078N-G118R-T213A, T022A-S024R-T033S-N116L-G118R-S128I, G020R-S078D-G118D-S188D-Q245R, S024F-S101G-S103A-V104I-N116A-Y209A-T213A-A232V, S024R-N062E-S078G-G118D-G159D-S188D, T022A-N062E-N116L-G118R-A158E-S166D, G020K-T022L-S078N-S166D-L217E, G020R-S078D-G118D-G159D, T022A-S024R-T033S-N116L, G020R-S078D-G118D-P129E-G159D, G020R-S078D-G118D-G159D-H249R, S024R-N062E-S078G-G118S, S024R-S078D-G118D-S188D-Q245R-H249R, A016S-S024F-T033S-S101G-S103A-V104I-N116L-Y209V-T213A-A232V, G020R-N062E-S078D-G118D-P129E-G159D-Q245R, S024F-S101G-S103A-V104I-N116L-G118V-Y209V-G211Q-A232V, S024F-S166D-L217E, T022A-S024R-S166D, S024R-N062E-S078G-G118D-P129E-N248D, A016S-S101G-S103A-V104I-N116L-S128N-Y209V-A232V, A016S-T022A-S101G-S103A-V104I-N116A-Y209V-G211Q-A232V, S078D-G118S-G159D-N248D-H249R, S024R-S078G-G118D-P129E-G159D-S188D, T022A-S024R-N062E-N116L-G118R-S128I-S188D, S103A-S128N, T022L-S078N-G118R-S166D-T213A, S024F-T033S-S101G-S103A-V104I-N116L-S128N-Y209A-T213A-A232V, T022A-S024R-N076D-S101G-S103A-V104I-P129E-G159D-S188D-A232V-Q245R-N248D-E271F-A272V, S024R-S078D-G118S-S128N-P129E, S024R-S078G-G118S-P129E-G159D-S188D-Q245R-H249R, T022A-S024R-N076D-S101G-S103A-V104I-P129E-G159D-S188D-A232V-Q245R-N248D-A270V-E271F-A272V, T022Q-S101G-S103A-V104I-G159D-S188D-A232T-Q245T-N248D, S024R-T033S-N116L-A158E, S130E-N269K, S024R-T033S-N116L-S128I-A158E, A016V-G020R-S024R-S078D-G118D-P129E-Q245R-N248D, S099G-S101A-G102A-S103G-V104I, A016S-T022A-S101A-V104L-S128N-L148I, G020R-S024R-S078G-G118S-P129E-S188D, S259W, S259H, T022A-S101G-S103A-V104I-P129E-G159D-S188D-A232V-Q245R-N248D-E271F, N018D-S101G-S103A-V104I-N116A-Y209A-G211Q-T213A-A232V, G020K-S024F-S078N-S166D-T213A, N076D-S101A-S103A-V104I-S188D-A232V-Q245R, S078D-G118D-P129E-G159D-H249R, S024R-S078G-G118D-P129E-G159D-N248D, S024R-N062E-S101A-S103A-V104I-S188D-A232V-Q245R, E271R, G020R-S024R-S078D-G118D-P129E-G159D-S188D, T022A-S024R-N076D-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-A270V-E271F, G020R-N062E-S078D-G118S-G159D-S188D-Q245R-H249R, Q012R-S101G-S103A-V104I-N116L-S128N-Y209V-A232V, G020R-S078G-G118D-P129E-G159D-S188D-Q245R-N248D, G020R-N062E-S078G-G118S-P129E-G159D-S188D-Q245R-H249R, T022A-S101G-S103A-V104I-G118R-S128L-G159D-S188D-A232V-Q245R-N248D-E271F, G020R-S078D-G118D-G159D-S188D-Q245R-N248D, T022A-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F-A272V, G020R-S024R-N062E-S078D-G118S-P129E-S188D-Q245R-N248D-H249R, G020R-S078G-G118S-G159D-H249R, G020K-T022L-S024F-G118R-S128D-S166D, T022A-S024R-N062E-N116L-G118R-S128I-A158E-S166D, A016S-S103G-V104L, A016S-T033S-S101G-S103A-V104I-N116L-Y209A-A232V, G020R-S024R-N062E-S078D-G118S-P129E-S188D-N248D, G020K-T022L-S078N-G118R-S166D, G020K-T213A-L217E, S024R-N062E-S078D-G118S-P129E-G159D-S188D-Q245R-N248D-H249R, S078N-S166D-T213A-L217E, S078D-G118S-P129E-G159D-Q245R-H249R, T022A-N076D-S101G-S103A-V104I-P129E-G159D-S188D-A232V-Q245R-N248D-E271F-A272V, G020K-T022L-G118R-L217E, T022A-V104L-L111V, T022A-S101G-S103A-V104I-S128A-P129E-G159D-S188D-A232V-Q245R-N248D-E271F, S024R-N062E-S078G-G118S-P129E-S188D-N248D, G020R-S024R-S078D-G118D-G159D-S188D-N248D, T022L-S078N-L217E, G020R-S024R-S078G-G118S-S188D, S024F-S101G-S103A-V104I-N116L-Y209V-G211Q-T213A-A232V, T022A-N116L-G118R-A158E-S166D, G020R-S024R-S078G-G118S-P129E-S188D-Q245R-N248D-H249R, T022A-V104I-S128N, S024R-S078G-G118D-Q245R, N062E-S078G-G118S-P129E-G159D-S188D-H249R, T022A-S101G-S103A-V104I-I107V-G159D-S188D-A232V-Q245R-N248D-A270V-E271F-A272V, S024R-N062E-S078D-G118S-P129E-G159D-Q245R-N248D-H249R, T022A-T033S-G118R-A158E-S188D, T022A-S078N-G097A-S101N-S103A-V104I-S128A-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, 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T022A-S024R-N116L-G118R-S166D, S024R-N062E-S078D-G118D-P129E-G159D-Q245R-N248D-H249R, G020K-T022L-S024F-S078N-G118R-S128D-T213A, T022A-S024G-N076D-S078N-G097A-S101Q-S103A-V104I-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S024G-S101N-S103A-V104I-S128A-G159D-S188D-A232V-Q245R-N248D-E271F-T274I, G097P-S099A-V104L, G097P-S099A-S103A, T022A-S024G-S078N-G097S-S101Q-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-V104L-L148I, T022A-S024G-G097A-S101N-S103A-V104I-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S024R-S101G-S103A-V104I-P129E-G159S-S188D-A232V-Q245R-N248D-E271F, T022A-S024G-N076D-S078N-S101G-S103A-V104I-Q109G-G159D-S188D-A232V-Q245R-N248D-E271F, S024R-S078D-G118S-G159D-Q245R-H249R, T022W-N043R-R045T-S101A-P210I-T213A, S024G-N076D-S078N-S101N-H120Q-S128A-L217Q, G020R-S024R-G118R-P129E-G159D-H249R, G020R-S078R-G159D-S188D, P129E-S188D-Q245R-H249R, S078R-P129E-Q245R, S078R-P129E-N248D, T022A-S101Q-S103A-V104I-P129S-A232V-Q245R, S188D-N248D, S078R-G118R-G159D, G020R-S078G-G118S-Q245R-N248D-H249R, S024R-S078R-G118R-G159D, G020R-P129E-G159D-N248D, G020R-S024R-G159D-N248D, G097A-S128A-L217Q, S024R-P129E-G159D-N248D, T022K-S101A-S103N-V104L-A232T-Q245R, S078G-G118S-S188D-Q245R-H249R, S099T-S101G-G102A-S103A, G020R-G118R-G159D-Q245R, S024G-S078N-S101N-L217Q, S024G-S078N-S101N-S128A-L217Q-S256R, S078R-G118R-G159D-N248D, S078G-G118S-Q245R-H249R, G118R-P129E-N248D, G020K-T022L-Q245R, A098Q-S099T-S101G-S103A, G118R-H249R, S024R-G118R-N248D-H249R, G020R-S024R-S078G-G118S-Q245R-N248D-H249R, A098F-S099A-S101G-V104I, G020R-T022W-S024R-N043R-R045T-S101A-G211Q, A098Q-S099G-S101G-V104L, G020K-N062E-G097A-S101Q-Q109G-N116L-S188D-L217Q, G020K-N062E-S078N-G097A-S101Q-Q109G-N116L, G020K-N062E-G097A-S101Q-Q109N-N116L-L217Q, N043R-S078N-G102A-S128L-L217E-S240R, S078N-G097S-S101Q, N043R-N062E-S078N-S101D-S128L-S240R, S128L-L217E-E271L, G020K-S024G-N062E-G097S-S101N-Q109N-N116L-S128A-L217Q, G020K-N062E-N116L-S128L-T213A, S078N-G097A-S101G-Q109G-S188D-L217Q, S101D-S128L-L217E-S240R, G020K-S024G-N062E-G097A-S101Q-Q109G-N116L-S128A, G020K-N062E-S078N-G097S-S101N-Q109N-N116L-S128A-S188D-L217Q, G020K-N062E-G097S-S101Q-Q109G-N116L-S128A-S188D-L217Q, G020K-S024G-N062E-S078N-G097S-S101N-Q109N-N116L-S128A, S024G-S078N-G097S-S101G-S128A-S188D-L217Q, S024G-S078N-G097A-S101N-Q109G-S128A-S188D-L217Q, G020K-N043R-G102A-S128L, G020K-S078N-S101D-G102A-L217E-S240R, G020K-N062E-G100S, S078N-S128L-L217E-E271L, S128Q-L217E-E271L, G020K-S024G-N062E-S078N-G097A-S101N-N116L-S128A-L217Q, G020K-S024G-N062E-S078N-G097S-S101G-N116L-S128A-L217Q, G020K-S078N-S101D-G102A-L217E, G020K-N062E-S128L, G020K-N062E-S078N-S101G-Q109N-N116L, G020K-S024G-N062E-S078N-G097S-S101Q-N116L-S128A-L217Q, S078N-S128L-L217E-S240R, S101D-S128Q-L217E-S240R-E271L, N043R-G102A-S240R, G020K-N062E-S078N-S101N-Q109N-N116L-M222S, G020K-N062E-S101N-Q109G-N116L-S128A-L217Q, G020K-S024G-N062E-S078N-G097A-S101G-N116L-L217Q, G020K-N062E-S101G-N116L-S188D, G020K-N062E-G097S-S101Q-Q109N-N116L-S128A-S188D, G020K-I035V-G100S-S128L-L217E, N043R-S101D-G102A-S240R, N062E-S078N-G100S, G020K-N043R-N062E-A215T, N062E-S240R, N043R-S128L-L217E-S240R, N062E-S101D-S240R, G020K-S101D-N116L-T213A, G020K-S101D-N116L-S128Q-P210S-T213A, G020K-S101D-T213A, G020K-S024G-N062E-S078N-G097S-S101Q-Q109N-N116L-S128A, G020K-N062E-S078N-G097S-S101Q-N116L-S128A-L217Q, S024G-G097S-S101N-Q109G-S188D-L217Q, G020K-S024G-N062E-S101N-Q109G-N116L-L217Q, L217E-S240R, S101D-L217E-S240R-E271L, N043R-N062E-S101D-S128L-S240R, G097A-S101N-S128A-S188D-L217Q, G020K-N062E-S078N-S101N-N116L-S128A-L217Q, G020K-N062E-S078N-G097S-S101G-Q109N-N116L-L217Q, G020K-S024G-N062E-G097S-S101N-Q109G-N116L-S128A-L217Q, G020K-S024G-N062E-G097S-S101G-Q109G-N116L-S128A-S188D-L217Q, G020K-N062E-S101N-Q109N-N116L-S128A-S188D, S078N-G097S-S101N-Q109N-S128A-S188D-L217Q-W241L, G020K-N043R-N062E-S078N-S101D, G020K-N062E-S101D-S240R, G020K-S101D-S128Q, G020K-N116L-S128Q-Y209H-T213A, G020K-N043R-S078N-G100S-L217E-N218D, S024G-S101N-Q109G-S128A-S188D-L217Q, G020K-S024G-N062E-S078N-G097S-S101N-Q109G-N116L-S128A-S188D, S078N-S101N-Q109G-S188D-L217Q, G020K-S024G-N062E-S078N-G097S-S101N-Q109G-N116L-S188D-L217Q, G020K-N062E-S078N-S101Q-Q109N-N116L-S128A-S188D-L217Q, G020K-N062E-S101G-Q109G-N116L, G020K-S024G-N062E-G097S-S101N-Q109G-N116L-S128A-S188D-M222S, S024G-G097A-S101G-S188D-L217Q, S024G-S078N-G097A-S101G-S128A-S188D, G097S-S101N-Q109G-S128A-S188D, G020K-S078N-S101D-I246T, N043R-S128L-L217E, G020K-S101D-L217E-S240R, S128L-L217E-T224A-E271L, G020K-S078N-S101D-S128Q-L217E-E271L, N062E-S078N-S101D-L217E-S240R, G020K-S101D-L217E-E271L, G020K-S078N-S128Q-L217E-S240R, G020K-N062E-S101D-N116L-T213A, G100S-S101D, S078N-G097S-S101G-S128A-L217Q, G020K-N062E-G097S-S101N-N116L-S128A-L217Q, G020K-N062E-G097S-S101N-N116L-L217Q, S101N-Q109G-S188D, G020K-S024G-N062E-G097S-S101Q-Q109N-N116L-S128A-L217Q, G020K-S024G-N062E-G097S-S101N-N116L, G020K-S024G-N062E-S101N-N116L-S128A, G020K-S024G-N062E-S078N-S101Q-Q109N-N116L-S128A, G020K-N062E-S078N-S101N-Q109G-N116L-S188D, G020K-N062E-S078N-S101N-Q109G-N116L-S128A, G020K-S024G-N062E-S078N-G097S-S101N-Q109G-N116L-S128A-M222S, G020K-S024G-N062E-S078N-G097S-S101G-N116L-S128A, S024G-S078N-G097S-S101Q-Q109G-S128A-S188D-L217Q, S024G-S078N-S101G-Q109N-S188D-L217Q, S024G-S078N-G097A-S101G-Q109G-S188D, S078N-S101D-S128Q-L217E-S240R-E271L, G020K-S128Q-L217E, S128Q-L217E-S240R-E271L, G020K-S078N-S101D-L217E, G020K-S078N-S101D-L217E-E271L, G020K-N116L-S128Q-T213A, G020K-N116L-N123G, S101G-S188D-L217Q, G020K-S024G-N062E-S078N-S101G-N116L-L217Q, S078N-G097A-S101G-Q109N-S128A-S188D, G020K-S078N-S128L-L217E-E271L, G020K-N062E-S101Q-Q109G-N116L, G020K-S101D-L217E, S024G-G097A-S101N-Q109N-S128A, G020K-S078N-S101D-S128L-L217E-S240R, G097A-S101G-L217Q, S024G-S101Q-Q109G-S128A-L217Q, G020K-S024G-N062E-T071A-S078N-G097S-S101G-N116L-L217Q, G020K-S024G-N062E-S078N-S101G-N116L-S128A, G020K-S024G-N062E-S078N-G097A-S101N-N116L, G020K-N062E-G097A-S101N-N116L, G020K-S024G-N062E-G097A-S101N-Q109G-N116L-S128A, G020K-S024G-N062E-S078N-G097S-S101G-Q109N-N116L, G020K-N062E-S078N-G097S-S101Q-Q109G-N116L-S128A-S188D-L217Q, G020K-N062E-S101N-Q109G-N116L-L217Q, G020K-S024G-N062E-G097A-S101N-Q109G-N116L-S188D-L217Q, G020K-S024G-N062E-S101G-N116L, S078N-G097S-S101N-Q109G-G159D-L217Q, S024G-G097A-S101N-Q109G-S188D, S078N-S101G-S128A-L217Q, S078N-G097A-S101N-S128A-S188D-L217Q, S024G-G097S-S101G-S128A-S188D-L217Q, S024G-S078N-G097S-S101Q-Q109G-S128A-S188D-L217Q-V227A, G020K-N062E-S078N, S078N-S128L-L217E-S240R-E271L, G020K-N062E-I072N-S101D-N116L-S128Q, G020K-T057P-N062E-N116L-S128Q-T213A, G020K-N043R-S078N-S101D-S240R, S078N-G097S-S101N-Q109N-S128A-S188D-L217Q, S024G-S101N-Q109N-S128A-S188D, G020K-S024G-N062E-G097S-S101Q-N116L-S128A, S024G-S078N-G097S-S101N-Q109N-S128A-S188D, N043R-G100S, G020K-N062E-S078N-S101Q-Q109G-N116L-S128A, S078N-S101Q-Q109N-S188D-L217Q, G020K-N116L-S128L-T213A, G020K-S078N-S101D-L217E-S240R, G020K-N062E-S101N-Q109N-N116L-M222S, G020K-S024G-N062E-G097A-S101Q-Q109N-N116L-S128A, S078N-G097S-S101G-S128A-S188D, G097S-S101G-Q109N-S128A-M222S, S024G-S078N-G097A-S101Q-S128A-M222S, G097A-S101G-Q109N-L217Q, S024G-S078N-S101G-S128A-S188D-L217Q, S024G-G097A-S101N-Q109G-S128A-S188D, G097S-S101N-Q109G-S128A, S078N-G097A-S101Q-L217Q, G020K-N043R-S078N-S101D, G020K-N043R-E054D-N062E-S078N-S240R, G020K-S101D-S240R, S128Q-L217E-S240R, G020K-S128Q-L217E-V244A-E271L, G020K-L217E-E271L, G020K-S078N-S128L-L217E, S078N-S101D-S128Q-L217E-E271L, G020K-N062E-L217E-E271L, G020K-N062E-S101D-N116L-S128Q-T213A, G020K-N062E-S078N-N116L-S128I, G020K-N062E-S078N-S128Q-T213A, S024G-G097A-S101G-Q109G-S128A, S024G-G097S-S101N-Q109N-S128A-L217Q, G020K-S024G-N062E-S078N-S101N-N116L-S128A-S188D-L217Q, G020K-S101D, G020K-N062E-S078N-G097S-S101Q-N116L-S128A, G020K-N062E-G097S-S101G-N116L-S188D, N043R-S078N-G100S-S128L-S240R, G097S-S101N-Q109N-S188D, G097S-S101Q-Q109N-S128A-L217Q, G020K-N062E-S101N-Q109G-N116L-S128A-S188D-M222S, G020K-S024G-N062E-S078N-G097S-S101Q-Q109G-N116L, G020K-N062E-M222S, G097S-S101Q-Q109G-S188D, G020K-S078N-L217E-S240R, G020K-N062E-S078N-G097A-S101N-Q109G-N116L-S188D, G020K-S024G-N062E-G097S-S101Q-Q109G-N116L-S188D, G020K-N062E-S078N-G097A-S101Q-N116L-S128A-M222S, G020K-N062E-S078N-S101N-N116L-S188D, G020K-S024G-N062E-S078N-S101G-N116L, G020K-N062E-S078N-S101N-N116L-S128A-S188D, G020K-S024G-N062E-G097S-S101N-Q109G-N116L-S128A-S188D-L217Q, G020K-N062E-G097S-S101N-Q109N-N116L, G020K-N062E-S101G-N116L-S188D-L217Q, G097A-S101N-S128A, S101G-Q109N-S128A-M222S, S024G-G097S-S101Q-Q109G-S188D-L217Q, G097S-S101Q-Q109N-S128A-S188D, S024G-S078N-G097A-S101Q-S188D-L217Q, S024G-S078N-S101G-Q109G-S128A-L217Q, G020K-N062E-S128L-S240R, N062E-S078N-S128L-S240R, N043R-G102A-S128L-L217E-S240R, N062E-S078N-L217E-S240R-E271L, S078N-S128Q-L217E-N269S, G020K-N062E-N123G-T213A, G020K-N062E-S078N-S099G-N116L-S128Q-T213A-L217N, G020K-N062E-S078N-S128I-L217N, G097A-S101Q-S128A-S188D-L217Q, G020K-N062E-S078N-G097S-S101N-Q109G-N116L, G020K-S024G-N062E-S078N-G097S-S101N-Q109G-N116L-S188D, G020K-N043R-S128L-S240R, S024G-S078N-G097A-S101N-Q109N-S128A, G020K-N062E-G097S-S101Q-N116L-L217Q, G020K-N062E-S078N-S128I, G020K-S024G-N062E-L090V-G097A-S101Q-N116L-M222S, G020K-N062E-S078N-S101Q-Q109G-N116L-S188D-L217Q, G020K-S024G-N062E-G097A-S101G-N116L-S128A-S188D-L217Q, G020K-N062E-G097S-S101N-Q109N-N116L-S188D-L217Q, G020K-S024G-A048V-N062E-S078N-G097A-S101N-N116L-S128A-L217Q, G020K-N062E-S078N-S101Q-Q109G-N116L, G020K-S024G-N062E-G097S-S101N-Q109N-N116L-S128A, G020K-N062E-S078N-G097A-S101Q-Q109G-N116L-S188D, G020K-N062E-S078N-G097A-S101N-Q109N-N116L-N218D, S024G-S101N-Q109G-S128A-S188D-M222S, S024G-G097A-S101Q-Q109N-S188D-L217Q, S024G-S078N-G097S-S101Q-Q109N-S128A-S188D, S101N-Q109G-M222S, S024G-I072V-G097S-S101N-S128A-L217Q, S101G-S128A, N043R-S078N-G102A-A215G, G020K-S101D-S128L-L217E-S240R, S078N-S101D-S128Q-L217E-S240R, G020K-N116L-S128L-M222S, G020K-N062E-S078N-N116L-T213A-M222S, G020K-N062E-S078N-N116L-S128Q, G020K-N062E-T213A или G020K-N062E-S078N-N116L-N123G-M222S , где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
13. Выделенный вариант субтилизина, где указанный вариант субтилизина представляет собой зрелую форму, обладающую протеолитической активностью, и содержит аминокислотную последовательность, включающую комбинацию замен аминокислот, выбранную из T022A-S024G-N076D-G097S-S101G-S103A-V104I-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, G020K-T022L-S078N-G118R-S128D-T213A, S024R-S078G-G118S-P129E-S188D-Q245R-H249R, T022A-N076D-S101G-S103A-V104I-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-S101G-S103A-V104I-S128A-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-S024G-G097S-S101Q-S103A-V104I-Q109G-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-N076D-S101N-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, T033S-G118R-S128I-A158E, T022A-S103A-L111V-L148I, T022A-N076D-A085T-S101G-S103A-V104I-P129E-S188D-A232V-Q245R-N248D-E271F, T022A-S024R-N076D-S101G-S103A-V104I-A158E-S188D-A232V-Q245R-N248D-E271F, G020K-T022L-S078N-S128D-T213A-L217E, T022A-T033S-P129E-S166D, T022A-S024G-N076D-G097A-S101Q-S103A-V104I-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-G097A-S101N-S103A-V104I-Q109G-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, G020K-S078N-S166D-T213A, T022A-S101Q-S103A-V104I-L124V-P129S-G159D-S188D-A232V-Q245R-N248D-E271F, T022Q-S101G-S103A-V104I-G159D-S188E-A232V-Q245T-N248D, A158E-S166D-S188D, L021S-T022A-S024G-N076D-S101Q-S103A-V104I-S128A-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-G097A-S101N-S103A-V104I-L124V-S128A-P129E-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-S101G-S103A-V104I-G159D-S188D-A232V-N238Y-Q245R-N248D-A270V-E271F, T022A-T033S-N062E-N116L-S188D, T022A-S024G-S049N-N076D-S101G-S103A-V104I-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-G097S-S101Q-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, G097P-A098Q-S099A-S103G-V104I, G020K-S024F-G118R-S128D-S166D, T022A-S024G-S101G-S103A-V104I-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S024R-T033S-G118R-S166D, T022A-S024R-N076D-S101G-S103A-V104I-G159D-S188D-A232V-N238Y-Q245R-N248D-A270V-E271F, N062E-S078D-G118S-P129E-N248D-H249R, T022A-S024G-V026I-S101N-S103A-V104I-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, G097P-A098Q-S099A-S101A, S078G-G118D-P129E-S188D-Q245R-H249R, T022A-G097A-S101N-S103A-V104I-L126I-P129E-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, G020R-S024R-N062E-S078G-G118D-P129E-G159D-S188D-Q236R-Q245R-N248D, T022A-N076D-S078N-S101N-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-N076D-S101G-S103A-V104I-P129E-G159D-S188D-A232V-N238Y-Q245R-N248D-E271F-A272V, T022A-N076D-N077D-S078N-G097S-S101N-S103A-V104I-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-N043R-N076D-S101G-S103A-V104I-P129E-A158E-S188D-A232V-Q245R-N248D-A270V-E271F, S099G-S101A-G102A-S103A, T022A-S024G-G097A-S101N-S103A-V104I-Q109G-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, G020R-S024R-S078D-G118S-P129E-Q245R, S056C, S024R-T033S-N062E-N116L, T022A-S024G-N076D-S101G-S103A-V104I-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T033S-S101G-S103A-V104I-N116L-S128N-Y209A-T213A-A232V, A098Q-S099A-G100S-S101G-V104I, T022A-S078N-S101N-S103A-V104I-Q109G-S128A-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-S024R-N076D-S101G-S103A-V104I-G159D-S188D-Q245R-N248D-A270V-E271F-A272V, T022A-N076D-G097S-S101Q-S103A-V104I-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S024G-S078N-S101N-S103A-V104I-Q109N-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-L090I-S101G-S103A-V104I-S128A-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, G020R-N062E-S078G-G118D-P129E-G159D-S188D-N248D-H249R, A016S-T033S-S101G-S103A-V104I-N116L-S128N-Y209V-T213A-A232V, G020R-S078G-G118D-N248D-H249R, T022A-N062Q-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, S024R-N062E-N116L-A158E-S166D-S188D, T022A-S024G-N076D-S101Q-S103A-V104I-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-S024R-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-A270V-E271F, T022A-N062Q-S101G-S103A-V104I-P129E-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-N076D-S101N-S103A-V104I-Q109G-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-S101G-S103A-V104I-P129E-S188D-A232V-Q245R-N248D-E271F, T022A-S024G-N076D-S078N-G097A-S101N-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S024R-N076D-S101G-S103A-V104I-I107V-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S024R-N116L-G118R, T022A-S024R-T033S-N062E-N116L-G118R-S128I-A158E-S188D, T022A-N076D-G097S-S101N-S103A-V104I-Q109G-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S024G-N076D-G097S-S101Q-S103A-V104I-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, S024R-T033S-N062E-N116L-S128I-S188D, T022A-S024G-N076D-G097S-S101N-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, S240V, N062E-G118R, T022A-S101G-S103A-V104I-G159D-S188E-A232V-Q245T-N248E, S024R-N062E-S078G-G118D-P129E-S188D-Q245R-N248D, A016S-S103A-V104I, A088P, T022A-S101N-S103A-V104I-Q109N-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-S101N-S103A-V104I-L126I-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, G097P-A098F-V104L, T022A-G097S-S101N-S103A-V104I-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-G097A-S101N-S103A-V104I-Q109N-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, G020K-T022L-S128D, T022A-S024G-S101G-S103A-V104I-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, G020R-S024R-N062E-S078D-G118D-P129E-G159D-S188D-N248D-H249R, S024F-S078N-G118R-L217E, T022A-S024G-S078N-G097S-S101Q-S103A-V104I-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S024R-S101G-S103A-V104I-A158E-G159E-S188D-A232V-Q245R-N248D-E271F-A272V, S024R-N062E-N116L-G118R-S128I-S188D, G118R-S128D-S166D-T213A, T022A-G097A-S101G-S103A-V104I-P129E-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-T033S-S101G-S103A-V104I-N116L-S128L-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S078N-S101Q-S103A-V104I-Q109G-S128A-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-G097S-S101N-S103A-V104I-Q109G-S128A-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-S024G-G097S-S101N-S103A-V104I-Q109N-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-N076D-G097A-S101N-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S101G-S103A-V104I-L126I-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, G020R-S078G-G118S-S188D-H249R, T022A-S024G-N076D-S101Q-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S101A-S103A, T022A-N076D-S101G-S103A-V104I-P129E-G159E-S188D-A232V-Q245R-N248D, S024R-S078D-G118D-S188D-N248D-H249R, S099A-V104I, T022A-G097S-S101Q-S103A-V104I-Q109N-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, S024R-S078G-G118D-G159D-N248D, T022A-S024G-S078N-S101Q-S103A-V104I-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-N076D-S101G-S103A-V104I-P129E-G159E-S188D-A232V-Q245R-N248D-E271F, T022A-G097A-S101Q-S103A-V104I-L124V-S128A-P129E-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-G097A-S101Q-S103A-V104I-L124V-G159D-S188D-A232V-Q245R-N248D-E271F, S078G-G118D-P129E, S212W, T022A-S101G-S103A-V104I-I107V-P129E-G159D-S188D-A232V-N238Y-Q245R-N248D-A270V-E271F-A272V, T022A-S078N-S101Q-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-G097A-S101G-S103A-V104I-S128A-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, S101G-V104L, T022A-S101G-S103A-V104I-I107V-P129E-G159D-S188D-A232V-N238Y-Q245R-N248D-E271F, T022A-S024R-S101G-S103A-V104I-P129E-G159D-S188D-A232V-Q245R-N248D-E271F-A272V, T022A-S024G-G097A-S101Q-S103A-V104I-Q109N-S128A-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-S024G-N076D-S101G-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S024R-N076D-S101G-S103A-V104I-P129E-A158E-S188D-A232V-Q245R-N248D-E271F-A272V, T022A-T033S-S101G-S103A-V104I-N116L-N117S-Y209A-G211Q-T213A-A232V, S101G-S103A-V104I-N116L-G118V-Y209A-G211Q-A232V, T022A-N062E-S101N-S103A-V104I-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S024G-G097S-S101N-S103A-V104I-S128A-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, S024F-T213A, T022A-S024G-N076D-S101N-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-N076D-S101Q-S103A-V104I-Q109G-S128A-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-S078N-G097A-S099G-S101G-S103A-V104I-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-N076D-S101G-S103A-V104I-G159D-S188D-L217Q-N218S-A232V-Q245R-N248D-E271F, T022A-S024G-N076D-S101N-S103A-V104I-Q109G-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-S024G-S101N-S103A-V104I-Q109N-G159D-S188D-A232V-Q245R-N248D-E271F, G020R-S024R-S078D-G118D-Q245R, T022A-S078N-I079V-S101N-S103A-V104I-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-S024R-N116L-G118R-S128I-A158E-S166D, S078G-G118D-S188D, G097P-A098Q-S099G-G102A, S099A-S103G, S078D-G118D-P129E-Q245R-N248D, T022A-S024G-S078N-S101Q-S103A-V104I-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-S078N-S101N-S103A-V104I-Q109G-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-N076D-S078N-G097S-S101Q-S103A-V104I-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-S024R-N116L-S128I-A158E-S166D-S188D, A016S-T022A-S101A-S103G-V104I, T022A-G097S-S101N-S103A-V104I-S128A-G159D-R186H-S188D-A232V-Q245R-N248D-E271F, T022A-G097A-S101Q-S103A-V104I-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-S101N-S103A-V104I-L126I-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-S101Q-S103A-V104I-L126I-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-N076D-G097S-S101N-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, T022A-N043R-N076D-S101G-S103A-V104I-P129E-S188D-A232V-Q245R-N248D-E271F-A272V, T022A-S024R-T033S-N062E-N116L-S128I-S188D, T022A-S024G-S078N-G097S-S101G-S103A-V104I-S128A-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022L-S078N-G118R-S128D-T213A, T022A-N076D-G097A-A098T-S101Q-S103A-V104I-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, T022A-S024G-S078N-G097A-S101Q-S103A-V104I-S128A-G159D-S188D-A232V-Q245R-N248D-E271F, S103A-V104L, S024R-S078G-G118D-P129E-G159D-S188D-N248D, T022A-S024R-N043R-S101G-S103A-V104I-S188D-A232V-Q245R-N248D-E271F, T022A-S024R-G061W-N076D-S101G-S103A-V104I-P129E-S188D-A232V-Q245R-N248D-E271F, G020R-N062E-S078D-G118S-S188D-N248D, T022A-S101G-S103A-V104I-A158E-G159E-S188D-A232V-Q245R-N248D-E271F, G100S-V104I, T022A-S024G-N076D-S078N-G097S-S101N-S103A-V104I-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, G097P-A098F-S103A, T022A-G097S-S101Q-S103A-V104I-Q109G-G159D-S188D-A232V-Q245R-N248D-E271F, S024F-S078N-G118R-S128D-L217E, T022A-S101Q-S103A-V104I-Q109G-S128A-G159D-S188D-L217Q-A232V-Q245R-N248D-E271F, S024R-T033S-N116L-S128I-A158E-S166D, T022L-S024F-S128D, T022A-S101G-S103A-V104L, T022A-N043R-S101G-S103A-V104I-A158E-G159E-S188D-A232V-Q245R-N248D-E271F-A272V, T022A-N076D-S101G-S103A-V104I-I107V-G159D-S188D-A232V-Q245R-N248D-E271F, T022L-S024F-S078N, T022A-N062E-A158E-S188D, G020K-T022L-S166D-T213A, T022A-S024G-T071A-S078N-S101N-S103A-V104I-G159D-S188D-A232V-Q245R-N248D-E271F, A016S-T022A-S101G-S103A-V104I-L148I, S078D-G118S-S188D-Q245R-N248D-H249R, G020K-T022L-S128D-S166D, 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G020K-S024G-N062E-S101Q-Q109N-N116L-S188D, G020K-N062E-S078N-N116L-S128I-L217N, S024G-S078N-G097A-S101N-Q109G-S188D, S024G-G097A-S101Q-Q109N-L217Q, G020K-S101D-S128L, S101N-Q109G-S128A, S078N-S101D-L217E, S024G-S078N-S101N-S128A-M222S, G020K-S024G-N062E-G097A-S101Q-N116L-S128A-S188D, S024G-S078N-G097A-S101N-Q109N-S128A-M222S, G020K-N062E-S078N-G097A-S101N-N116L-S128A-S188D-L217Q, G020K-S024G-N062E-S078N-G097S-S101Q-N116L-S188D, G020K-N062E-S078N-G097S-S101G-N116L-M222S, G020K-S024G-N062E-S078N-S101N-N116L-M222S, G020K-S024G-N062E-G097S-S101Q-N116L, G020K-S024G-N062E-S101Q-Q109G-N116L-S128A-S188D, G020K-S024G-N062E-G097S-S101N-Q109N-N116L-S128A-S188D-L217Q, G020K-N062E-S078N-G097S-S101N-N116L, S024G-S078N-S101G-Q109G-M222S, S078N-G097A-S101G-Q109G-S128A, S078N-G097S-S101N-Q109N-S128A, S078N-G097A-S101N-Q109N-S188D, S024G-S101N-S128A-S188D-M222S, S101Q-Q109N-S188D-M222S, S101D-G102A-L217E, G020K-N062E-S078N-S101D-L217E, G020K-S078N-S101D-I107T-S128L-S240R, G020K-S078N-G100S-L217E-N218D, G020K-N043R-S101D, N043R-G100S-S128L-L217E, G020K-S078N-S101D-S128L-L217E-E271L, G020K-S101D-S128L-E271L, G020K-S101D-N116L-S128L-T213A, G020K-N062E-S078N-T213A-L217N, G020K-N062E-N116L-L217N, G020K-N062E-S128I-T213A, N062E-S101D-S128L-S240R, G097S-S101N-S128A, G020K-N062E-S078N-N116L-T213A-L217N, G020K-S101D-M222S, S024G-S101G-Q109G, G097A-S101Q-Q109G, G097S-S101Q-Q109N-S188D, G020K-N062E-G097S-S101N-N116L-S128A-S188D-L217Q, S078N-S101Q-S128A-M222S, S078N-G097A-S101Q-Q109N, S024G-S078N-G097A-S101Q-S128A-S188D, S078N-S101D-G102A, G020K-N062E-S078N-G097S-S101Q-Q109G-N116L-S128A-S188D-M222S, G020K-S024G-N062E-S078N-G097S-S101N-Q109N-N116L-S188D-L217Q-M222S, G020K-N062E-G097S-S101Q-N116L-S128A, G020K-S024G-N062E-G097S-S101Q-N116L-S128A-S188D-L217Q, S024G-S101N-Q109N-S128A, S024G-G097S-S101Q-S128A-S188D, S024G-S101Q-L217Q, G097S-S101G-S128A, S024G-S078N-S101Q-S128A-L217Q, S024G-S078N-G097S-S101N-S188D, S024G-G097A-S101N-S128A-M222S, G097A-S101Q-Q109N-S188D-M222S, S024G-S078N-G097S-S101Q, G020K-S101D-S128L-S212P-L217E-S240R, G020K-S101D-S128L-L217E-N218S-S240R, S101D-S128Q, G020K-N062E-S078N-E271L, S101D-E271L, S101D-S240R-E271L, G020K-S101D-E271L, G020K-N116L-S128L-T213A-M222S, G020K-N116L-S128Q-M222S, G020K-N062E-V084A-S128Q-M222S, G020K-N062E-S078N-N116L, S078N-S101N-Q109G-S128A-M222S, S024G-G097S-S101Q-Q109G, G020K-N062E-T213A-L217N, G097S-S101G-S128A-S188D, G020K-S240R, N062E-S078N, G020K-S128Q-M222S, S078N-G097S-S101G-Q109G, G020R-S024G-N043R-S078N-S101Q, G097S-S101N-Q109G-S128A-M222S, S024G-G097S-S101G-Q109N, S024G-S078N-G097A-S101G-Q109N, G020K-N062E-S078N-S101N-N116L-S128A-S188D-L217Q, G020K-N062E-S101Q-Q109G-N116L-S128A-M222S, G020K-S024G-N062E-S101Q-Q109N-N116L, S024G-S078N-S101Q-M222S, S024G-G097A-S101Q-Q109G-S128A, S101Q-Q109N-L217Q, S101Q-Q109G-S188D-L217Q, S024G-G097A-S101N-S188D-M222S, S024G-S078N-G097A-S101Q-Q109N, G097A-S101Q-S128A, G097A-S101Q-Q109G-S128A, S024G-G097S-S101Q-M222S, S078N-S101D-L217E-S240R, G020K-G100S-S128L, G020K-N043R-S078N-S128L-S240R, G020K-N043R-S101D-S240R, G020K-N043R-S078N-S128L, N062E-S128L, G020K-S078N-S101D-S128L, S078N-S101D, G020K-S101D-N116L-T213A-M222S, G020K-T213A-M222S, G020K-N116L-T213A, G020K-N116L, G020K-N062E-S128Q-L217N, S078N-G097S-S101G-Q109N-S128A, S078N-S101Q-Q109G, N062E-S078N-S128L, G020K-N062E-N116L-S128Q-T213A-L217N, G020K-N116L-T213A-M222S, G020K-S024G-N062E-S101Q-Q109H-N116L-M222S, G020K-N116L-S128Q-T213A-M222S, S024G-G097S-S101N, G020K-P055T-N062E-N116L-L217N, S024G-S101Q-S128A-S188D-M222S, G020K-M222S, G020K-N062E-S078N-G097A-S101N-N116L-S188D-L217Q, G097S-S101G-Q109N-M222S, G020K-S024G-N062E-S078N-G097A-S101N-N116L-S128A-L217Q-M222S, G020K-S024G-N062E-G097S-S101N-Q109N-N116L-S128A-S188D-L217Q-M222S, G020K-N062E-S078N-S101Q-N116L-S128A, S024G-S078N-S101N-Q109G-S188D-L217Q-M222S, S101N-Q109N-S128A-M222S, S101Q-Q109G-S128A-S188D, G097A-S101G-S188D-M222S, S078N-G097S-S101N-Q109G-S128A-S188D-L217Q-M222S, S024G-G097A-S101Q-M222S, S024G-G097S-S101N-Q109N-S128A-M222S, S024G-S078N-G097S-S101N-M222S, G020K-G102A-S240R, S078N-S128Q-E271L, G020K-N062E-S078N-S128Q, G020K-N062E-S078N-T213A-M222S, G020K-N062E-S078N-S128I-T213A-L217N, S078N-G097A-S101N, G020K-N062E-N116L-S128I-T213A-L217N, S024G-G097A-S101Q-S128A-S188D-M222S, S024G-S078N-G097A-S101N, S024G-S078N-G097S-S101G-M222S, G020K-N062E-S078N-N116L-S128Q-T213A-M222S, G020K-N062E-S128Q-M222S, G020K-N062E-S101Q-N116L, G020K-N062E-S101Q-Q109G-N116L-S188D-L217Q, G020K-S024G-N062E-G097A-S101N-N116L-S128A-S188D, G020K-S024G-N062E-S078N-S101Q-Q109G-N116L-S188D-M222S, S078N-G097S-S101N-Q109G-S128A-L217Q-M222S, S024G-S078N-S101N-Q109G-S128A-L217Q-M222S, G097A-S101Q-S128A-S188D, S024G-S078N-G097A-S101N-Q109N-S128A-S188D-L217Q-M222S, S024G-S078N-G097A-S101G-L217E-M222S, S078N-G097S-S101Q-Q109G-T274I, G097A-S101G-M222S, S101N-Q109N, G020K-L217E-S240R-E271L, S128Q-S240R-E271L, G020K-S101D-N116L-S128Q-M222S, G020K-P055L-N062E-N116L-S128I, G020K-N062E-S101G-N116L-S128A-M222S, G020K-S078N-S101D-S128L-S240R-E271L, G020K-N062E-S078N-G097S-S101Q-Q109N-N116L-L217Q-M222S, S078N-S101D-S240R-E271L, S024G-S078N-S101N-Q109G-L217Q-M222S, G097A-S101G-Q109G-L217Q-M222S, G020K-S024G-N062E-S078N-G097S-S101N-N116L-S128A-L217Q-M222S, G020K-S078N-S128L-E271L, G020K-N062E-S078N-N116L-L217N, G097A-S101G-Q109G-S188D-L217Q-M222S, S024G-S078N-G097S-S101N-Q109G-S128A, G020K-S024G-N062E-S078N-G097S-S101N-N116L-S188D-M222S, G020K-S024G-N062E-G097S-S101Q-N116L-S128A-M222S, G020K-N062E-S078N-G097A-S101G-Q109G-N116L-S128A-S188D-L217Q, G020K-N062E-S101G-N116L-S128A-L217Q-M222S, S024G-G097A-S101Q-S188D-M222S, S024G-S101G, I072V-S101Q-S128A, S078N-G097S-S101N-Q109N, S024G-G097A-S101N-Q109N-S188D-L217Q-M222S, S078N-S101Q-Q109N-S188D-L217Q-M222S, G020K-N043R-N062E-S128L-L217E, N043R-G102A-S128L-S240R, G020K-N116L-N123G-T213A-M222S, G020K-S101D-S128Q-T213A-M222S, S024G-G097A-S101Q-Q109G, G020K-S078N, G020K-N062E-S101Q-N116L-S188D, G020K-N062E-S078N-G097S-S101N-Q109G-N116L-S188D-L217Q-M222S, G020K-S024G-N062E-G097S-S101G-N116L-S128A-L217Q-M222S, G020K-N062E-G097A-S101N-N116L-S128A-L217Q-M222S, S078N-G097S-S101G-Q109N-M222S, S024G-G097A-S101Q-Q109N, S024G-G097S-S101N-Q109N-S128A-S188D-L217Q-M222S, S078N-S101N-Q109N-S188D-L217Q-M222S, S078N-S101D-S128L-L217E-S240R, G020K-N062E-S101D-N116L-S128L, G097S-S101N-S188D-M222S, G020K-S078N-S128L-S240R-E271L, G020K-N062E-N116L-S128I-T213A, G020K-S024G-N062E-S078N-G097S-S101Q-N116L-S188D-M222S, G020K-N062E-S101Q-Q109G-N116L-L217Q-M222S, G020K-S024G-N062E-G097A-S101Q-N116L-L217Q-M222S, G020K-N062E-S078N-G097S-S101Q-N116L-S128A-S188D-M222S-A232T, G020K-S024G-N062E-S078N-G097S-S101Q-Q109N-N116L-S128A-L217Q-M222S, G020K-N062E-G097A-S101Q-N116L-S188D-M222S, S024G-S078N-G097S-S101N-Q109G-S128A-L217Q-M222S, S024G-G097A-S101Q-Q109N-S128A-S188D-L217Q-M222S, G020K-N043R, G020K-N043R-S078N-G102A-S128L, G097R-S101D, G020K-N062E-S078N-I107V-N123G-S128I, G020K-N062E-S078N-N116L-N123G-T213A, G020K-N062E-N116L-S128I-M222S, G020K-N043R-S240R, S078N-S101D-S128L, G020K-S078N-S128Q-E271L, S078N-S101G-S128A-S188D-L217Q-M222S, S078N-G097S-S101G-Q109N-S128A-L217Q-M222S или G020K-N062E-N116L-S128Q-T213A-M222S, где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
14. Выделенный вариант субтилизина, где указанный вариант субтилизина представляет собой зрелую форму, обладающую протеолитической активностью, и содержит аминокислотную последовательность, включающую комбинацию замен аминокислот, выбранную из G020R-T022W-S024R-R045T-S101A-N204S-G211Q, T022W-S024R-S101A-P210I-T213A, G020R-S101A-G211Q, G020R-S024R-S101A-P210I-G211Q-T213A, G020R-S101A-P210I-T213A, T022W-S024R-N043R-S101A-P210I, G020R-S024R-N043R-R045T-S101A-G211Q, T022R-S101T-S103N-V104L-A232M-Q245R, G020R-T022A-S024R-R045T-S101A-T213A, T022R-S101G-S103N-V104I-A232M-Q245R, G020R-T022W-S024R-S101A-N116L-G211Q-T213A, G020R-T022W-N043R-S101A-N116L-G211Q, G020R-T022W-N043R-R045T-S101A, T022W-N043R-R045T-S101A-P210I-T213A, G020R-S024R-R045T-S101A-Q109R-P210I-G211Q-T213A, S024R-N043K-S101A-N204D-P210I, G020R-S024R-R045T-S101A-P210I, S024R-N043R-R045T-S101A-P210I-T213A, G020R-S024R-R045T-S101A-T213A, T022R-S101A-S103N-V104I-A232T-Q245R, G020R-T022W-S024R-R045T-S101A-N204S-P210I-G211Q, T022W-N043R-S101A-P210I-G211Q, G020R-S024R-N043K-R045T-S101A-N116L-P210I, T022A-S101G-S103A-V104I-I107V-L217Q-A232V-Q245R, S024R-N043R-R045T-S101A-G211Q-T213A, T022R-S101A-S103G-V104I-A232M-Q245R, T022R-S101G-S103N-V104L-A232V-Q245R, S024R-S078G-G118S-Q245R-N248D-H249R, S024R-N043R-S101A-P210I-T213A, T022K-S101N-S103A-V104I-A232T-Q245R, G020R-N043R-S101A-P210I-G211Q, T022R-S101T-S103A-V104I-A232M-Q245R, S024R-V026A-N043R-S101A-P210I-G211Q, T022A-S078N-S101G-S103A-V104I-S128A-L217Q-A232V-Q245R, T022K-S101T-S103G-V104I-A232L-Q245R, G020R-T022W-S024R-N043R-R045T-S101A-G211Q, G020R-S024R-S078G-G118D-Q245R, T022A-S101N-S103A-V104I-S128A-P129S-A232V-Q245R, T022A-G097A-S101G-S103A-V104I-L217Q-A232V-Q245R, G020R-T022W-N043R-S101A-G211Q-T213A, N018K-G020R-S024R-R045T-S101A-P210I-G211Q-T213A, G020R-T022W-S024R-N043R-R045T-S101A, T022R-S101N-S103A-V104L-A232T-Q245R, G020R-S024R-S101A-T213A, T022R-S101N-S103G-V104I-A232V-Q245R, G020R-T022W-S024R-S101A-G211Q-T213A, T022A-S024G-S101G-S103A-V104I-S128A-L217Q-A232V-Q245R, S024R-S078G-G118D-P129E-Q245R-H249R, T022R-S101T-S103A-V104L-A232L-Q245R, G020R-S024R-S101A-G211Q-T213A, T022A-S024G-S101G-S103A-V104I-L217Q-A232V-Q245R, G020R-T022W-N043R-V051A-S101A-P210I-G211Q, S024R-S078G-G118D-Q245R, N043R-S101A-P210I, T022Y-S101G-S103N-V104I-A232L-Q245R, G020R-T022W-N043R-S101A-T213A, G020R-S024R-S078D-G118S-S188D-Q245R-H249R, T022Y-S101N-S103N-V104I-A232T-Q245R, G020R-T022W-S024R-F050L-S101A-G211Q-T213A, N018S-T022W-S024R-N043R-S101A-G211Q, T022A-S101G-S103A-V104I-S128A-P129S-L217Q-A232V-Q245R, T022R-S101N-S103N-V104I-A232V-Q245R, T022A-S024G-S078N-G097S-S101Q-S103A-V104I-L217Q-A232V-Q245R, S101A-P210I-G211Q-T213A, G020R-N043R-R045T-S101A-P210I-G211Q-T213A, T022R-S101A-S103N-V104I-A232M-Q245S, G020R-T022W-N043R-R045T-S101A-G211Q, G020R-T022W-N043R-R045T-S101A-N116L-P210I, G020R-S024R-S078D-G118S-Q245R, G020R-T022W-S024R-N043R-S101A-P210I-G211Q-T213A, G020R-S024R-N043R-R045T-V051A-S101A-P210I-G211Q, T022A-S101G-S103A-V104I-P129S-L217Q-A232V-Q245R, G020R-S078G-G118D-Q245R-N248D-H249R, T022K-S101A-S103N-V104L-A232T-Q245R, T022A-N076D-S078N-G097S-S101G-S103A-V104I-L217Q-A232V-Q245R, S024R-N043R-R045T-S101A-P210I, T022A-S024G-G061R-S078N-S101G-S103A-V104I-S128A-A232V-Q245R, T022A-S078N-S101G-S103A-V104I-L217Q-A232V-Q245R, T022A-S024G-S078N-S101Q-S103A-V104I-S128A-L217Q-A232V-Q245R, T022Q-S101T-S103A-V104L-A232V-Q245R, G020R-T022W-S024R-R045T-S101A-G211Q, P014R, T022R-S101A-S103N-V104L-A232M-Q245R, T022K-S101G-S103N-V104I-A232V-Q245R, G020R-S024R-S078D-G118D-Q245R, T022A-G097S-S101N-S103A-V104I-L217Q-A232V-Q245R, G020R-S024R-S101A-P210I-T213A, T022K-S101N-S103N-V104I-A232V-Q245R, T022Y-S101T-S103A-V104I-A232L-Q245R, T022A-N076D-S078N-S101G-S103A-V104I-S128A-L217Q-A232V-Q245R, G020R-S024R-N043R-R045T-S101A-P210I-G211Q-T213A, G020R-R045T-S101A-P210I-T213A, G020R-S078D-G118S-Q245R-H249R, G020R-T022W-R045T-V051A-S101A, T022R-S101T-S103A-V104I-A232V-Q245R, N043R-S101A, G020R-S024R-N043R-R045T-S101A-G211Q-T213A, T022R-S101G-S103G-V104I-A232L-Q245R, G020R-S101A-P210I, T022K-S101T-S103N-V104I-A232V-Q245R, G020R-T022W-N043R-V051A-S101A, G020R-T022W-N043R-S101A, G020R-T022W-S024R-N043R-S101A-P210I, G020R-T022W-S024R-N043R-R045T-S101A-P210I-T213A, G020R-N043R-R045T-S101A-T213A, T022A-S024G-S078N-S101N-S103A-V104I-L217Q-A232V-Q245R, T022Y-S101N-S103N-V104L-A232T-Q245R, G020R-S101A-N116L-P210I-G211Q, T022R-S101A-S103N-V104L-A232L-Q245R, G020R-S024R-N043R-S101A, T022R-S101G-S103A-V104L-A232T-Q245R, G020R-S024R-S101A-G211Q, T022A-S101Q-S103A-V104I-P129S-A232V-Q245R, T022A-S024G-G097A-S101G-S103A-V104I-L217Q-A232V-Q245R, T022A-S024G-S078N-S101G-S103A-V104I-S128A-L217Q-A232V-Q245R, T022K-S101A-S103N-V104I-A232T-Q245R, G020R-T022W-S024R-R045T-S101A, T022Y-S101G-S103A-V104I-A232V-Q245R, S024R-N043R-R045T-S101A-N116L-P210I-G211Q-T213A, T022A-S024G-S078N-S101N-S103A-V104I-S128A-L217Q-A232V-Q245R, G020R-N043R-S101A-P210I, 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T022A-S101G-S103A-V104I-A232V-N238Y-Q245R, G020R-S078G-G118D-P129E-G159D-Q245R-H249R, S024R-S078D-G118S-S188D-Q245R-H249R, T022A-S101G-S103A-V104I-A232V-Q245R-A270V, T022A-S024G-G097A-S101Q-S103A-V104I-A232V-Q245R, T022A-S101T-S103G-V104I-A232V-Q245R, G020R-T022W-S101A-T213A, T022A-S101G-S103A-V104I-S128A-A232V-Q245R, T022A-S101N-S103A-V104I-Q109G-A232V-Q245R, T022Q-S101N-S103N-V104L-A232L-Q245R, N018S-G020R-S101A-P210I-G211Q, G020R-S078D-G118S-S188D-Q245R-H249R, T022A-S101G-S103N-V104I-A232L-Q245R, T022R-S101T-S103G-V104L-A232V-Q245R, T022A-N076D-S101G-S103A-V104I-A232V-Q245R-A270V-A272V, T022Q-S101N-S103G-V104I-A232V-Q245S, G020R-N043R-R045T-S101A-G211Q, T022A-N076D-S101G-S103A-V104I-L217Q-A232V-Q245R, T022R-S101A-S103A-V104L-A232V-Q245S, T022A-S078N-S101Q-S103A-V104I-Q109N-A232V-Q245R, S024R-S078G-G118S-T180N-Q245R-N248D, V004M, T022A-S101T-S103N-V104I-A232T-Q245R, T022A-S078N-S101G-S103A-V104I-A230T-A232V-Q245R, T022Y-S101N-S103N-V104I-A232L-Q245R, G020R-S024R-S078D-G118S-G159D-Q245R-N248D, G020R-S024R-S078G-G118S-G159D-S188D-Q245R, G020R-N043R-R045T-S101A-P210I-T213A, S024R-S078D-G118D-Q245R-H249R, G020R-S024R-N043R-R045T-S101A-Q109R-T213A, T022A-S078N-S101N-S103A-V104I-Q109N-A232V-Q245R, G020R-S024R-N043R-S101A-N204D-G211Q-T213A, T022A-G097A-S101Q-S103A-V104I-A232V-Q245R, G020R-S024R-S078G-G118S-N248D, G020R-S024R-S078G-G118S-G159D-Q245R-N248D, T022A-S024G-S078N-S101N-S103A-V104I-S128A-A232V-Q245R, S024R-S078G-G118D-G159D-Q245R-H249R, T022K-S101T-S103N-V104L-A232V-Q245R, S024R-S078G-G118S-Q245R, T022A-S101A-S103G-V104I-A232L-Q245R, T022A-G097A-S101Q-S103A-V104I-S128A-A232V-Q245R, T022A-S024G-S078N-G097S-S101G-S103A-V104I-A232V-Q245R, G020R-S078D-G118S-Q245R, T022A-S024R-N076D-S101G-S103A-V104I-A232V-N238Y-Q245R-A270V-A272V, S101A-P210I-T213A, T022A-S101Q-S103A-V104I-S128A-P129S-L217Q-A232V-Q245R, A114L, A016S-T022A-S101G-S103A-V104I-N116A-Y209V-G211Q-T213A-A232V, T022A-S101N-S103A-V104I-S128A-P129S-L217Q-A232V-Q245R, T022R-S101A-S103N-V104I-A232L-Q245S, T022Q-S101N-S103A-V104I-A232M-Q245W, T022W-S024R-S101A-N204S-G211Q-T213A, T022A-S101N-S103A-V104L-A232M-Q245R, T022A-S024G-S101Q-S103A-V104I-Q109N-S128A-A232V-Q245R, G020R-S078D-G118S-P129E-Q245R-H249R, T022Q-S101T-S103A-V104I-A232T-Q245S, S024R-N043R-R045T-S101A-P210I-G211Q-T213A, T022R-S101N-S103G-V104I-A232T-Q245R, S024R-S101A-P210I-G211Q-T213A, T022A-S024G-N076D-G097A-S101G-S103A-V104I-S128A-A232V-Q245R, G020R-S024R-S078G-G118S-S188D-N248D-H249R, T022A-G097S-S101Q-S103A-V104I-Q109G-S128A-A232V-Q245R, G020R-S024R-R045T-S101A-P210I-T213A, T022A-S078N-G097S-S101N-S103A-V104I-S128A-A232V-Q245R, T022A-S024G-S078N-G097A-S101Q-S103A-V104I-A232V-Q245R, T022R-S101G-S103N-V104I-A232T-Q245W, T022K-S101G-S103A-V104I-A232V-Q245R, T022W-S024R-S101A-G211Q-T213A, S024R-N043R-S101A-P210I-G211Q, T022Q-S101T-S103G-V104I-A232T-Q245R, T022A-S101G-S103A-V104I-A232V-Q245R-A272V, T022K-S101G-S103N-V104L-A232M-Q245R, T022A-S024G-G097A-S101Q-S103A-V104I-Q109N-A232V-Q245R, S078D-G118S-G159D-Q245R-H249R, G020R-T022W-S024R-A069T-S101A-P210I-T213A, T022A-S101N-S103A-V104I-S128A-L217Q-A232V-Q245R, G020R-T022W-S024R-N043R-R045T-S101A-P210I-G211Q, G020R-S024R-S078D-G118D-G159D-S188D-Q245R-H249R, T022A-S078N-S101N-S103A-V104I-S128A-L217Q-A232V-Q245R, G020R-S078G-G118D-G159D-Q245R-N248D-H249R, T022W-R045T-S101A-T213A, G020R-S024R-S078N-G097A-S101A-Q109N-N116L-L217Q, T022R-S024F-S101G-S103A-V104I-A232V-Q245R-E271H, G020R-S024R-S078N-G097S-S101Q-S128A-L217Q, T022R-S101G-S103A-V104I-A232V-Q245R-E271H, G020R-S024R-S101G-N116L-S128A, G020R-S024R-S101A-Q109N-N116L-S128A, T022R-S024F-S101G-S103A-V104I-A232V-Q245R-T260M, G020R-S024R-S078N-S101N-N116L-S128A, G020R-S024R-G097A-S101A-Q109G-N116L, G020R-S024R-S101N-S128A, G020R-S024R-S078N-S101G-S128A-L217Q, T022R-S024F-S101G-S103A-V104I-A232V-Q245R, T022R-S078N-G097S-S101Q-S103A-V104I-Q109N-N116L-A232V-Q245R, G020R-S024R-G097A-S101A-Q109N-N116L-S128A, G020R-S024R-G097S-S101N-Q109N-S128A, T022R-S101G-S103A-V104I-A232V-P239G-Q245R, G020R-S024R-G097S-S101A-S128A, G020R-S024R-S101G-Q109N-N116L-A215V, G020R-S024R-S078N-G097S-S101Q-Q109G-N116L-S128A, G020R-S024R-S078N-G097S-S101N-Q109N, G020R-S024R-S101G-S128A, T022R-S101G-S103A-V104I-A232V-Q245R-T260K-L267N, T022R-S101G-S103A-V104I-S128A-A232V-Q245R, T022R-S024G-G097S-S101N-S103A-V104I-A232V-Q245R, T022R-S024F-S101G-S103A-V104I-V121F-A232V-Q245R, G020R-S024R-G097A-S101Q-N116L-S128A-L217Q, T022R-G097A-S101N-S103A-V104I-N116L-A232V-Q245R, G020R-S024R-S078N-G097S-S101A-N116L-S128A, G020R-S024R-G097A-S101G-S128A-L217Q, T022R-S078N-S101G-S103A-V104I-N116L-A232V-Q245R, G020R-S024G-N043R-S078N-S101Q-Q109G-N116L-S128A-Q206R-L217Q, G020R-S024R-S078N-G097S-S101Q-N116L-S128A, G020R-S024R-S078N-G097S-S101N-Q109N-N116L-S128A, G020R-S024R-S101N-Q109G-N116L, G020R-S024R-S101Q-N116L-S128A-L217Q, G020R-N043R-S078N-S101G-Q109G-N116L, G020R-S024G-N043R-S078N-G097S-S101G-S128A, T022R-S024G-S078N-S101N-S103A-V104I-S128A-A232V-Q245R, T022R-S024G-G097A-S101Q-S103A-V104I-Q109G-N116L-S128A-A232V-Q245R, T022R-S101G-S103A-V104I-A232V-P239G-Q245R-E271I, G020R-S024R-S101Q-S128A, G020R-S024R-G097A-S101N-Q109G-N116L, G020R-S024G-N043R-S078N-G097A-S101G-Q109G-N116L, G020R-S024R-G097S-S101Q-Q109N-N116L-L217Q, T022R-S024G-S078N-G097A-S101Q-S103A-V104I-A232V-Q245R, G020R-S024R-G097A-S101G-Q109N-S128A, G020R-S024R-G097S-S101A-Q109N-S128A, T022R-G097S-S101Q-S103A-V104I-Q109G-A232V-Q245R, G020R-S024R-S078N-G097A-S101N-S128A-L217Q, G020R-S024G-N043R-S078N-G097S-S101G-N116L, G020R-S024R-S078N-G097S-S101G-Q109N-N116L-L217Q, T022R-S024G-S078N-S101N-S103A-V104I-Q109N-N116L-S128A-A232V-Q245R, G020R-S024R-S078N-G097S-S101A-Q109N-S128A-L217Q, G020R-S024R-S078N-G097A-S101G-Q109N-N116L-L217Q, G020R-S024G-N043R-S078N-S101Q, G020R-S024R-S078N-G097A-S101Q-Q109G-N116L, G020R-N043R-G097A-S101N-N116L-S128A, G020R-S024R-S078N-S101G-Q109G-N116L-S128A-L217Q, G020R-S024R-S078N-S101Q-Q109G-N116L-S128A-L217Q, G020R-S024R-S078N-G097S-S101G-Q109N-N116L-S128A, G020R-S024R-S078N-G097S-S101N-Q109N-N116L, T022R-S101N-S103A-V104I-Q109N-N116L-S128A-A232V-Q245R, T022R-S024G-G097S-S101G-S103A-V104I-Q109G-N116L-S128A-A232V-Q245R, T022R-S024G-S078N-S101G-S103A-V104I-N116L-S128A-A232V-Q245R, G020R-S024R-S078N-S101N-Q109N-N116L, G020R-S024R-S078N-S101A-Q109N-N116L-S128A, T022R-S024G-G097A-S101Q-S103A-V104I-N116L-S128A-A232V-Q245R, T022R-G097S-S101G-S103A-V104I-S128A-A232V-Q245R, T022R-G097A-S101Q-S103A-V104I-S128A-A232V-Q245R, T022R-S024G-S078N-G097S-S101Q-S103A-V104I-S128A-A232V-Q245R, T022R-S078N-G097A-S101Q-S103A-V104I-N116L-S128A-A232V-Q245R, G020R-S024R-S101G-Q109G-N116L-L217Q, G020R-S024R-S078N-S101Q-Q109N-N116L, G020R-S024R-S101G-N116L-S128A-L217Q, G020R-S024R-S101A-Q109G-N116L-S128A, T022R-S024G-S078N-G097A-S101G-S103A-V104I-N116L-L217Q-A232V-Q245R, T022R-S024G-S078N-G097S-S101Q-S103A-V104I-Q109G-N116L-S128A-A232V-Q245R, G020R-N043R-G097A-S101N-S128A-L233I, T022R-S101Q-S103A-V104I-A232V-Q245R, G020R-N043R-G097S-S101Q-Q109G-N116L-S128A, T022R-S078N-S101Q-S103A-V104I-A232V-Q245R, G020R-S024R-G097S-S101N-Q109G-N116L, T022R-S078N-G097S-S101G-S103A-V104I-S128A-A232V-Q245R, T022R-S024G-S101N-S103A-V104I-A232V-Q245R, G020R-S024R-G097A-S101N-Q109G-S128A, G020R-N043R-S101Q-N116L-S128A-L217Q, G020R-S024R-S078N-G097A-S101Q-Q109N-S128A, G020R-S024R-E089D-S101A-Q109N-N116L, G020R-S024R-G097A-S101G-Q109G-S128A, G020R-S024G-N043R-S101G-N116L-S128A-L217Q, G020R-N043R-S078N-S101Q-Q109G-L217Q, G020R-S024G-N043R-G097A-S101N-Q109N-N116L-L217Q, G020R-S024R-S078N-G097A-S101G-N116L-S128A, G020R-S024R-G097S-S101A-Q109N-N116L, G020R-S024R-S078N-S101A-Q109G-S128A, G020R-S024R-G097A-S101A-Q109G-N116L-S128A, G020R-N043R-G097A-S101Q-N116L-S128A, G020R-N043R-G097S-S101Q-S128A-L217Q, G020R-S024R-S078N-G097A-S101G-Q109G-N116L-L217Q, G020R-N043R-G097A-S101G-Q109N-N116L-S128A, G020R-N043R-S078N-G097S-S101N-Q109N-L217Q, G020R-S024R-G097S-A098E-S101A-N116L-S128A, G020R-S024R-S078N-G097S-S101A-Q109G-S128A, T022R-S024G-S078N-G097S-S101G-S103A-V104I-Q109N-N116L-L217Q-A232V-Q245R, G020R-S024R-S078N-G097A-S101Q-Q109N-N116L-L217Q, T022R-S024G-G097S-S101Q-S103A-V104I-S128A-A232V-Q245R, G020R-S024R-S101A-Q109N-N116L-S128A-L217Q, G020R-S024R-S101G-Q109G-N116L-S128A-L217Q, T022R-S078N-G097A-S101N-S103A-V104I-S128A-A232V-Q245R, G020R-S024G-N043R-S101N-N116L-S128A-L217Q, G020R-S024R-S078N-G097S-S101A-Q109N-N116L, G020R-S024R-S078N-G097A-S101N-Q109N-N116L-S128A, G020R-S024R-S078N-S101Q-Q109N-N116L-L217Q, T022R-S024G-S078N-S101N-S103A-V104I-N116L-S128A-A232V-Q245R, G020R-N043R-S101G-Q109N-N116L-S128A-L217Q, T022R-S101N-S103A-V104I-L217Q-A232V-Q245R, T022R-G097A-S101N-S103A-V104I-N116L-S128A-A232V-Q245R, G020R-S024R-G097A-S101A-S128A-L217Q, T022R-S024G-S078N-G097A-S101N-S103A-V104I-S128A-L217Q-A232V-Q245R, G020R-S024R-S078N-G097S-S101N-Q109G-N116L-S128A-L217Q, T022R-S024G-S101G-S103A-V104I-N116L-L217Q-A232V-Q245R, T022R-G097A-S101Q-S103A-V104I-N116L-S128A-A232V-Q245R, G020R-N043R-S101Q-S128A-L217Q, G020R-S024R-S078N-S101N-Q109G-S128A-L217Q, G020R-S024R-S078N-G097A-S101N-Q109N-N116L-S128A-L217Q, T022R-S024G-G097A-S101Q-S103A-V104I-N116L-L217Q-A232V-Q245R, G020R-S024R-S078N-G097S-S101G-S128A-N204S-L217Q, T022R-S024G-S078N-S101Q-S103A-V104I-N116L-S128A-A232V-Q245R, G020R-S024R-S078N-G097S-S101Q-Q109N-N116L-L217Q, G020R-S024R-S078N-S099G-S101Q-Q109N-N116L-L217Q, T022R-S024G-G097A-S101G-S103A-V104I-Q109G-N116L-L217Q-A232V-Q245R, T022R-S078N-G097A-S101Q-S103A-V104I-S128A-L217Q-A232V-Q245R, T022R-G097A-S101G-S103A-V104I-Q109G-N116L-L217Q-A232V-Q245R, G020R-S024R-G097A-S101N-Q109N-S128A-L217Q, T022R-G097S-S101N-S103A-V104I-S128A-L217Q-A232V-Q245R, G020R-S024R-K094R-S101N-N116L-S128A-L217Q, T022R-S024G-S078N-G097A-S101G-S103A-V104I-N116L-S128A-L217Q-A232V-Q245R, G020R-S024R-G097A-S101A-N116L-S128A-L217Q, G020R-S024R-S078N-G097W-S101Q-Q109N-N116L-L217Q, T022R-G097S-S101N-S103A-V104I-N116L-S128A-L217Q-A232V-Q245R, T022R-S101Q-S103A-V104I-N116L-L217Q-A232V-Q245R, T022R-S024G-S078N-G097A-S101Q-S103A-V104I-N116L-S128A-L217Q-A232V-Q245R, T022R-S078N-G097A-S101Q-S103A-V104I-Q109N-N116L-S128A-L217Q-A232V-Q245R, G020R-N043R-G097S-S101Q-Q109G-N116L-S128A-L217Q, T022R-S024G-G097A-S101G-S103A-V104I-N116L-S128A-L217Q-A232V-Q245R, T022R-G097S-S101G-S103A-V104I-L217Q-A232V-Q245R, T022R-S024G-G097A-S101Q-S103A-V104I-Q109G-N116L-L217Q-A232V-Q245R, T022R-S024G-S078N-G097A-S101Q-S103A-V104I-S128A-L217Q-A232V-Q245R, T022R-S101Q-S103A-V104I-Q109G-N116L-S128A-L217Q-A232V-Q245R, T022R-S078N-G097S-S101G-S103A-V104I-N116L-S128A-L217Q-A232V-Q245R, T022R-S024G-T071A-S078N-S101N-S103A-V104I-Q109N-N116L-S128A-A232V-Q245R, T022R-S024G-G097A-S101Q-S103A-V104I-S128A-L217Q-A232V-Q245R, T022R-S101G-S103A-V104I-Q109N-S128A-A232V-Q245R, T022R-S024G-G097S-S101G-S103A-V104I-S128A-L217Q-A232V-Q245R, T022R-S024G-S078N-S101Q-S103A-V104I-Q109N-N116L-S128A-L217Q-A232V-Q245R, T022R-S101G-S103A-A232V-Q245R, T022R-S101G-S103A-V104I-A232V-Q245R-E271A, T022R-S103A-V104I-A232V-Q245R, T022R-S101G-V104I-A232V-Q245R, T022R-S101G-S103A-V104I-Q245R, G020R-N043R-E271A, G020R-S024R-I035T-S101A-N116L, G020R-N043R-P239G, G020R-N043R-S242L или G020R-N043R-V234F, где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
15. Выделенный вариант субтилизина, где указанный вариант субтилизина представляет собой зрелую форму, обладающую протеолитической активностью, и содержит аминокислотную последовательность, включающую комбинацию замен аминокислот, выбранную из T022Y-S101N-S103A-V104I-A232T-Q245R, G020R-R045T-S101A-P210I-G211Q-L267F, T022Y-S101N-S103N-V104L-A232M-Q245R, T022A-P052T-S101G-S103A-V104I-L217Q-A232V-Q245R, T022A-S101G-S103N-V104L-A232T-Q245R, T022Y-S101A-S103N-V104L-A232V-Q245R, T022A-S101A-S103N-V104I-A232T-Q245R, T022Y-S101A-S103N-V104I-A232L-Q245R, T022Y-S101G-S103A-V104I-A232T-Q245R, T022A-S024G-N076D-S078N-G097S-S101N-S103A-V104I-L217Q-A232V-Q245R, T022A-S101N-S103N-V104L-A232M-Q245R, T022A-S101Q-S103A-V104I-S128A-P129S-A232V-Q245R, T022A-S101N-S103A-V104I-S128A-A232V-Q245R, S024R-S078D-G118S-G159D-Q245R-H249R, T022K-S101N-S103A-V104L-A232T-Q245R, T022R-S101T-S103N-V104I-A232T-Q245S, T022Q-S101N-S103N-V104I-A232M-Q245R, S024R-S078G-G118D-Q245R-H249R, T022R-S101N-S103N-V104L-A232V-Q245R, T022A-N076D-S101Q-S103A-V104I-Q109G-A232V-Q245R, G020R-S024R-S078G-G118D-S188D-Q245R-N248D, T022Y-S101N-S103A-V104I-A232V-Q245R, T022Q-S101N-S103A-V104L-A232L-Q245R, T022Q-S101N-S103N-V104I-A232L-Q245R, T022A-G097S-S101N-S103A-V104I-S128A-L217Q-A232V-Q245R, T022R-S101N-S103N-V104I-A232T-Q245S, G020R-S078D-G118D-G159D-Q245R-H249R, T022R-S101N-S103A-V104L-A232M-Q245R, T022A-N076D-G097S-S101Q-S103A-V104I-Q109N-S128A-A232V-Q245R, T022R-S101G-S103N-V104L-A232T-Q245S, G020R-T022W-N043R-R045T-S101A-G211Q-T213A, T022Q-S101T-S103N-V104I-A232T-Q245R, G020R-S024R-N043R-S101A-N204D-T213A, T022A-S024G-S101Q-S103A-V104I-Q109G-A232V-Q245R, G020R-S024R-S078D-G118D-S188D-Q245R-N248D-H249R, S024R-N043R-S101A-P210I, T022A-N076D-S101G-S103A-V104I-A232V-Q245R-A270V, G020R-S024R-S078D-G118S-H249R, S024R-S078G-G118S, T022R-S101G-S103N-V104L-A232V-Q245S, S024R-N043K-R045T-S101A, T022A-N076D-S078N-S101N-S103A-V104I-A232V-Q245R-T274A, T022A-S024G-N076D-S078N-S101G-S103A-V104I-L217Q-A232V-Q245R, T022Q-S101T-S103G-V104I-A232M-Q245W, S024R-S078G-G118S-P129E-Q245R-H249R, T022A-S024R-N076D-S101G-S103A-V104I-A232V-Q245R, V026W, T022R-S101G-S103N-V104I-A232L-Q245W, G020R-S024R-S078D-G118S-N248D, G020R-R045T-S101A-N204S-P210I, G020R-S024R-S078G-G118D-G159D-N248D-H249R, T022Y-S101A-S103N-V104I-A232V-Q245R, T022A-S024G-S078N-G097A-S101N-S103A-V104I-L217Q-A232V-Q245R, S024R-S101A-P210I-G211Q, T022R-S101N-S103N-V104I-A232V-Q245S, T022A-S024G-G097A-S101N-S103A-V104I-Q109G-A232V-Q245R, T022A-G097A-S101N-S103A-V104I-A232V-Q245R, T022A-S024G-N076D-S101Q-S103A-V104I-Q109N-A232V-Q245R, G020R-S078G-G118S-H249R, S078G-G118S-Q245R-H249R, T022A-G097A-S101N-S103A-V104I-L217Q-A232V-Q245R, T022Y-S101T-S103G-V104I-A232M-Q245R, T022K-S101G-S103N-V104I-A232L-Q245R, T022A-S024G-G097S-S101N-S103A-V104I-A232V-Q245R, S024R-S078G-G118D-G159D-S188D-Q245R-H249R, T022Q-S101N-S103A-V104I-A232V-Q245R, G020R-S024R-S078G-G118D-G159D-Q245R-N248D, T022A-S024G-S101G-S103A-V104I-A232V-Q245R, T022Q-S101G-S103A-V104L-A232T-Q245R, T022Q-S101T-S103A-V104I-A232T-Q245R, T022A-N076D-S101N-S103A-V104I-S128A-A232V-Q245R, T022A-S024G-N076D-S078N-G097A-S101G-S103A-V104I-A232V-Q245R, T022A-S101G-S103A-V104L-A232V-Q245R, T022A-S024G-S078N-G097S-S101Q-S103A-V104I-S128A-A232V-Q245R, T022A-S024G-N076D-G097S-S101G-S103A-V104I-A232V-Q245R, S024R-N043R-S101A-G211Q-T213A, T022A-G097S-S101Q-S103A-V104I-A232V-Q245R, S024R-S078G-G118S-Q245R-N248D, G020R-T022W-S024R-N043R-S101A-N204S-P210I-G211Q, T022A-S024G-S078N-G097S-S101N-S103A-V104I-Q109G-S128A-A232V-Q245R, T022A-S101G-S103A-V104I-P129S-A232V-Q245R, T022R-S101T-S103G-V104I-A232T-Q245S, T022A-S024G-I072V-G097S-S101Q-S103A-V104I-S128A-L217Q-A232V-Q245R, T022K-S101A-S103A-V104I-A232V-Q245W, G020R-T022W-S024R-R045T-S101A-N116L-P210I, T022R-S101T-S103G-V104I-A232T-Q245W, T022A-S024G-N076D-S078N-G097S-S101N-S103A-V104I-S128A-A232V-Q245R, T022A-G097A-S101N-S103A-V104I-S128A-A232V-Q245R, T022Y-S101T-S103N-V104L-A232V-Q245R, G020R-S024R-N062E-S078G-G118S-N248D, T022Q-S101G-S103A-V104I-A232V-Q245R, T022A-N076D-S078N-S101N-S103A-V104I-S128A-A232V-Q245R, T022A-S024G-G097A-S101N-S103A-V104I-S128A-A232V-Q245R, T022A-S024G-N076D-S078N-S101Q-S103A-V104I-A232V-Q245R, T022A-N076D-S078N-S101G-S103A-V104I-L217Q-A232V-Q245R, T022A-S024G-N076D-S101N-S103A-V104I-A232V-Q245R, T022R-S101N-S103A-V104I-A232L-Q245S, T022A-S078N-S101N-S103A-V104I-Q109N-L217Q-A232V-Q245R, T022R-S101A-S103A-V104I-A232V-Q245W, T022A-S101Q-S103A-V104I-A232V-Q245R, T022A-S078N-G097S-S101N-S103A-V104I-A232V-Q245R, T022A-G097A-S101Q-S103A-V104I-P129S-A232V-Q245R, T022A-S101T-S103A-V104I-A232L-Q245R, T022A-A085T-G097S-S101Q-S103A-V104I-L217Q-A232V-Q245R, T022A-S101T-S103N-V104L-S105G-A232M-Q245R, T022A-S101A-S103A-V104I-A232M-Q245S, T022Q-S101A-S103A-V104I-A232M-Q245R, T022Q-S101G-S103A-V104L-A232M-Q245R, T022A-S024G-G097A-S101Q-S103A-V104I-S128A-A232V-Q245R, T022A-S024G-G097S-S101Q-S103A-V104I-L217Q-A232V-Q245R, T022A-N076D-S078N-G097S-S101G-S103A-V104I-S128A-A232V-Q245R, T022A-G097A-S101G-S103A-V104I-H120Q-S128A-A232V-Q245R, T022A-G097A-S101N-S103A-V104I-S128A-P129S-L217Q-A232V-Q245R, G020R-S024R-N062E-S078G-G118D-Q245R, T022K-S101G-S103G-V104I-A232V-Q245R, S078G-G118S-S188D-Q245R-H249R, T022W-N043R-S101A-G211Q-T213A, T022A-S024G-N076D-G097S-S101N-S103A-V104I-Q109N-A232V-Q245R, T022Y-S101A-S103N-V104L-A232M-Q245R, G020R-T022W-S101A-N116L-G211Q-T213A, T022W-S024R-N043R-R045T-S101A-T213A, N043R-S101A-G211Q-T213A, T022Y-S101N-S103N-V104L-A232L-Q245R, T255G, T022A-G097S-S101N-S103A-V104I-S128A-A232V-Q245R, T022W-S024R-N043R-S101A-P210I-T213A, N269K, T022R-S101T-S103A-V104I-A232L-Q245S, T022A-S024G-I072V-S078N-G097S-S101N-S103A-V104I-S128A-A232V-Q245R, T022K-S101T-S103G-V104I-A232M-Q245R, H017R-N043R-R045T-S101A-P210I-T213A, T022A-S024G-G097S-S101G-S103A-V104I-S128A-A232V-Q245R, T022A-S078N-G097A-S101Q-S103A-V104I-A232V-Q245R, T022A-N076D-G097A-S101N-S103A-V104I-A232V-Q245R, T022A-S024G-V026A-S101G-S103A-V104I-S128A-A232V-Q245R, T022A-S024G-G097A-S101G-S103A-V104I-S128A-L217Q-A232V-Q245R, T022Q-S101T-S103G-V104I-A232L-Q245R, T022W-S024R-S101A-P210I-G211Q-T213A, T022Q-S101N-S103A-V104I-A232M-Q245R, G020R-R045T-S101A, T022Y-S101N-S103N-V104I-A232V-Q245R, T022R-S101G-S103A-V104I-A232L-Q245R, S024R-S078D-G118D-P129E-Q245R-H249R, T022R-S101A-S103A-V104I-A232M-Q245S, G020R-S101A-P210I-G211Q, T022W-N043R-R045T-S101A-P210I-G211Q, T022R-S101G-S103A-V104I-A232M-Q245S, T022A-S024G-N076D-S101G-S103A-V104I-L217Q-A232V-Q245R, T022R-S101T-S103N-V104I-A232V-Q245S, T022Q-S101G-S103N-V104L-A232M-Q245R, T022K-T038I-S101G-S103A-V104I-A232T-Q245S, N043R-R045T-S101A-P210I-G211Q, T022K-S101T-S103A-V104L-A232M-Q245R, T022Y-S101T-S103N-V104I-A232T-Q245R, G020R-S024R-N043R-S101A-P210I-T213A, S099G-S101G, T022A-S101N-S103N-V104L-A232T-Q245R, T022A-S024G-N076D-S078N-G097A-S101N-S103A-V104I-A232V-Q245R, T022R-S101N-S103A-V104I-A232V-Q245W, T022W-N043R-R045T-S101A-P210I, T022K-S101A-S103G-V104I-A232T-Q245R, T022Q-S101N-S103A-V104I-A232T-Q245S, T022A-S101Q-S103A-V104I-Q109G-A232V-Q245R, T022A-G097A-S101Q-S103A-V104I-S128A-P129S-L217Q-A232V-Q245R, T022A-S078N-G097A-S101Q-S103A-V104I-L217Q-A232V-Q245R, S024R-S078G-G118S-G159D-Q245R, T022K-S101N-S103N-V104I-A232T-Q245R, A098F-S099A-V104I, G020R-R045T-S101A-G211Q-T213A, T022A-N076D-S078N-G097A-S101G-S103A-V104I-A232V-Q245R, S024R-S078D-G118S-G159D-Q245R, T022A-S024G-N076D-S101N-S103A-V104I-Q109N-A232V-Q245R, T022K-S101T-S103N-V104I-A232M-Q245R, T022A-S024G-S078N-G097S-S101N-S103A-V104I-S128A-A232V-Q245R, T022Q-S101T-S103N-V104I-A232L-Q245S, A114T, T022Q-S101N-S103N-V104L-A232T-Q245R, T022A-S078N-S101G-S103A-V104I-Q109N-A232V-Q245R, T022K-S101T-S103A-V104I-A232M-Q245R, T022A-S024G-G097S-S101Q-S103A-V104I-Q109G-S128A-L217Q-A232V-Q245R, T022A-S101G-S103A-V104I-N116L-Y209V-G211Q-A232V, T022A-S101T-S103N-V104I-A232M-Q245W, G020R-T022W-S024R-S101A-N116L-G211Q, T022A-S078N-S101Q-S103A-V104I-S128A-L217Q-A232V-Q245R, G020R-R045T-S101A-G211Q, T022A-N076D-S078N-G097A-S101G-S103A-V104I-S128A-A232V-Q245R, S024R-S101A-T213A, T022A-S078N-S101G-S103A-V104I-A232V-Q245R, G020R-S024R-S078D-G118S-G159D-S188D-Q245R-N248D-H249R, T022Y-S101N-S103N-V104I-A232V-Q245W, T022Y-S101A-S103G-V104I-A232V-Q245R, T022A-S024R-N076D-S101G-S103A-V104I-A232V-N238Y-Q245R, G020R-S078G-G118S-G159D-H249R, T022A-S024R-S101G-S103A-V104I-P129E-A232V-Q245R-A272V, T022R-S101N-S103A-V104I-A232V-Q245S, G020R-S101A-P210I-G211Q-T213A, G020R-S024R-S078D-G118S-G159D-S188D-Q245R, S087R, G020R-S024R-R045T-A048T-S101A-G211Q, T022A-S024G-G097S-S101Q-S103A-V104I-A232V-Q245R, G020R-S024R-S078G-G118S-P129E-S188D-Q245R-N248D-H249R, T022A-G097S-S101G-S103A-V104I-Q109N-A232V-Q245R, L021S-T022A-S024G-N076D-S078N-S101G-S103A-V104I-S128A-A232V-Q245R, S242K, T022K-S101A-S103A-V104I-A232V-Q245R, T022K-S101N-S103A-V104L-A232L-Q245R, T022A-N076D-S101G-S103A-V104I-A232V-N238Y-Q245R-A270V-A272V, T022R-S101T-S103A-V104I-A232V-Q245S, T022Y-S101N-S103G-V104I-A232L-Q245R, T022Q-S101N-S103G-V104I-A232V-Q245R, T022A-S101G-S103A-V104I-A232V-Q245W, T022R-S101A-S103A-V104I-A232L-Q245S, G020R-T022W-R045T-F050L-S101A-P210I-G211Q-T213A, T022A-S101Q-S103A-V104I-S128A-L217Q-A232V-Q245R, G020R-S101A-G211Q-T213A, T022A-N076D-S101G-S103A-V104I-A232V-Q245R-A272V, T022A-S101G-S103A-V104I-A232V-N238Y-Q245R-A270V, T022A-S101N-S103G-V104I-A232V-Q245R, G020R-S101A-T208P-P210I-G211Q, D181S, S101G-S103A-V104I-N116L-Y209V-A232V, T022Q-S101A-S103N-V104I-A232T-Q245R, T022A-S101G-S103N-V104L-G211S-A232T-Q245R, T022A-G097A-S101Q-S103A-V104I-L217Q-A232V-Q245R, T022A-S101T-S103N-V104I-A232M-Q245S, T022A-S101G-S103A-V104I-Q109N-A232V-Q245R, T022Q-S101A-S103G-V104I-A232M-Q245R, G020R-S078G-G118S-G159D-N248D-H249R, T022A-N076D-S101N-S103A-V104I-A232V-Q245R, T022A-S024G-G097S-S101N-S103A-V104I-Q109G-S128A-A232V-Q245R, T022Q-S101A-S103A-V104I-A232T-Q245R, T022A-S101G-S103A-V104I-A232L-Q245R, T022A-S101G-S103A-V104I-P129E-A232V-N238Y-V244I-Q245R-A270V-A272V, G020R-S024R-S078D-G118D-G159D-Q245R-N248D-H249R, T022R-S101A-S103N-V104I-A232V-Q245W, T022Q-S101A-S103N-V104I-A232M-Q245R, T022Q-S101A-S103N-V104L-A232L-Q245R, T022A-S024G-S101Q-S103A-V104I-S128A-L217Q-A232V-Q245R, T022Q-S101G-S103G-V104I-A232M-Q245R, G020R-S078G-G118S-G159D-Q245R-H249R, T022R-S101A-S103N-V104L-A232V-Q245S, T022A-S024V-G097S-S101N-S103A-V104I-H120N-S128A-A232V-Q245R, T022A-A073V-N076D-S078N-S101N-S103A-V104I-A232V-Q245R, T022Q-S101G-S103N-V104L-A232V-Q245R-T274I, T022A-S101N-S103N-V104I-A232T-Q245R, T022W-S024R-R045P-S101A-P210I-G211Q-T213A, G020R-S024R-N062E-S078G-G118D-Q245R-N248D, T022Q-S101A-S103N-V104L-A232M-Q245R, T022Q-S101T-S103G-V104L-A232T-Q245R, T022R-S101G-S103N-A232L-Q245R, T022A-G097S-S101Q-S103A-V104I-Q109N-L217Q-A232V-Q245R, T022A-S024G-G097A-S101Q-S103A-V104I-S128A-L217Q-A232V-Q245R, S024R-S101A-G211Q-T213A, T022R-S101N-S103N-V104L-A232T-Q245S, G020R-R045T-S101A-P210I, T022R-S101A-S103A-V104I-A232M-Q245W, T022A-S101N-S103N-V104I-A232L-Q245R, T022A-S101G-S103G-V104I-A232M-Q245R, G020R-S024R-S078D-G118S-S188D-Q245R-N248D, S099T-S103G, T022W-N043R-S101A-P210I, T022W-A029V-N043R-S101A-P210I, F050R, T022A-S024G-N076D-S078N-G097S-S101Q-S103A-V104I-Q109N-A232V-Q245R, T022A-S024G-N076D-S101G-S103A-V104I-A232V-Q245R, T022R-S101G-S103N-V104L-A232L-Q245R, T022A-S101N-S103A-V104I-Q109N-S128A-A232V-Q245R, S024R-R045T-S101A, T022A-N076D-G097S-S101N-S103A-V104I-L217Q-A232V-Q245R, T022Q-S101G-S103A-V104I-A232M-Q245S, T022W-S024R-N043R-R045T-S101A-G211Q-T213A, G020R-S078G-G118S, T022W-N043R-G046C-S101A-P210I-G211Q, S101A-G211Q-T213A, T022A-N076D-S078N-S101G-S103A-V104I-A232V-Q245R, T022A-S024G-N076D-G097S-S101Q-S103A-V104I-A232V-Q245R, T022A-S101N-S103A-V104I-L126I-P129S-A232V-Q245R, T022R-S101T-S103N-V104I-A232L-Q245W, T022A-S101G-S103A-V104I-A232V-N238Y-Q245R-A270V-A272V, S078G-G118D-Q245R, T022A-S024R-S101G-S103A-V104I-P129E-A232V-N238Y-Q245R-A270V, T022A-N076D-G097S-S101N-S103A-V104I-Q109G-S128A-A232V-Q245R, T022W-S024R-R045T-S101A-N116L-P210I-T213A, T022R-S101T-S103N-V104I-A232L-Q245S, T022W-S024R-N043R-R045T-S101A-S106L-P210I-G211Q, T022A-E089G-G097A-S101Q-S103A-V104I-P129S-L217Q-A232V-Q245R, T022A-S101A-S103G-V104I-A232L-Q245S, N043R-S101A-P210I-T213A, N043R-R045T-S101A-T213A, S078G-G118D-H120N-H249R, T022Q-S101T-S103N-V104I-A232L-Q245R-R275H, T022Y-S101A-S103A-V104I-A232T-Q245S, T022K-S101N-S103N-V104I-A232M-Q245R, T022R-S101T-S103N-V104L-A232V-Q245S, T022A-N076D-G097S-S101N-S103A-V104I-A232V-Q245R, M119L, A098F-S099A-S101G-V104I, S078D-G118S-Q245R, T022Y-S101N-S103A-V104I-A232L-Q245S, T022A-G097A-S101N-S103A-V104I-S128A-L217Q-A232V-Q245R, G020R-S078G-G118D-N248D-H249R, T022R-S101A-S103N-V104I-A232T-Q245S, T022R-S101G-S103N-V104I-A232L-Q245S, S024R-S078D-G118D-S188D-Q245R-H249R, S132H, A098Q-S099G-S101G-V104L, N043R-S101A-P210I-G211Q-T213A, T022Q-S101G-S103A-V104I-A232M-Q245W, T022A-S101Q-S103A-V104I-L126I-P129S-A232V-Q245R, T022K-S101T-S103G-V104I-A232M-Q245S, T022W-N043R-R045T-S101A-G211Q-T213A, T022Q-S101N-S103G-V104I-A232V-Q245W, D181T, G020R-S024R-S078D-G118S-P129E-G159D-Q245R-H249R, G020R-S024R-S078G-G118D-G159D-S188D-Q245R, T022A-S101T-S103N-V104L-A232L-Q245S, T022A-S101G-S103N-V104L-A232V-Q245S, T022A-S024G-G097A-S101N-S103A-V104I-S128A-L217Q-A232V-Q245R, T022A-S024G-N076D-S101N-S103A-V104I-S128A-A232V-Q245R, T022Y-S101T-S103A-V104L-A232M-Q245R, T022Q-S101T-S103N-V104L-A232T-Q245R, F050S, T022Y-S101G-S103N-V104I-A232V-Q245R, G020R-R045T-S101A-T213A, T022K-S101N-S103G-V104I-A232V-Q245W, S024R-S078G-G118D-P129E-G159D-Q245R-H249R, T022Q-S101T-S103N-V104I-A232M-Q245W, S049W, T022A-S101A-S103N-V104L-A232L-Q245R, T022A-N076D-S101G-S103A-V104I-I107V-P129G-A232V-Q245R-A270V, S099A-V104I, T022Q-S101A-S103G-V104I-A232V-Q245W, T022A-N076D-S101G-S103A-V104I-A232V-N238Y-Q245R, T022A-S101G-S103A-V104I-L124V-L217Q-A232V-Q245R, T022R-G097S-S101Q-S103A-V104I-S128A-L217Q-A232V-Q245R, T022R-G097A-S101Q-S103A-V104I-Q109N-S128A-A232V-Q245R, R010A-T022R-S024F-S101G-S103A-V104I-A232V-Q245R-L267N-E271H, G020R-S024G-N043R-S078N-S101G-Q109N-N116L-L217Q-M222S, G020R-N043R-E271H, G020R-N043R-E271I, G020R-S024R-S101A-N116L-N269I, T022R-S101G-S103A-V104I-A232V-Q245R-E271F, G020R-N043R-E271F, G020R-N043R-N269I, G020R-S024R-S101A-N116L-E271H, T022R-S101G-S103A-V104I-A232V-Q245R-E271H, G020R-I035T-N043R, G020R-S024R-S101A-A114T-N116L, G020R-S024R-S101A-N116L-E271I, G020R-N043R-A114T, G020R-N043R-L267N, G020R-S024R-S101A-N116L-L267N, G020R-S024R-V028A-S101A-N116L, G020R-S024R-S101A-N116L-G258R, G020R-N043R-G258R, G020R-S024R-S101A-N116L-S242L, G020R-S024R-S101A-N116L-P239G, I008N-G020R-S024R-S101A-N116L, G020R-S024R-S101A-N116L-V234F, T022R-S101G-S103A-V104I-A232V, G020R-S024R-S101A-N116L-E271A, R010A-G020R-N043R, G020R-S024R-S101A-N116L-E271F, G020R-V028A-N043R, I008N-G020R-N043R или G020R-S024F-S101A-N116L, где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
16. Выделенный вариант субтилизина, где указанный вариант субтилизина представляет собой зрелую форму, обладающую протеолитической активностью, и содержит аминокислотную последовательность, включающую комбинацию замен аминокислот, выбранную из G020R-S024R-S078D-G118D-G159D-H249R, G020R-S024R-S078G-G118S-G159D-S188D-Q245R-N248D-H249R, T022A-S101T-S103G-V104I-A232T-Q245S, T022A-S101T-S103G-V104I-A232T-Q245R, T022Q-S101G-S103N-V104I-A232V-Q245W, T022W-N043R-S101A-T213A, T022A-N076D-S078N-G097A-S101N-S103A-V104I-S128A-A232V-Q245R, T022A-S024G-S078N-S101Q-S103A-V104I-S128A-A232V-Q245R, T022K-S101N-S103N-V104L-A232T-Q245R, T022A-S024G-S101Q-S103A-V104I-S128A-A232V-Q245R, G020R-S024R-N062E-S078G-G118D, G020R-S078G-G118D, T022Y-S101T-S103G-V104L-A232T-Q245R, R010H-T022A-S101G-S103A-V104I-A232V-Q245R-A272V, T022A-S024R-S101G-S103A-V104I-P129E-A232V-Q245R-A270V-A272V, T022A-N076D-S078N-G097S-S101Q-S103A-V104I-Q109G-A232V-Q245R, G020R-S024R-S078D-G118S-P129E-Q245R, T022A-N076D-S101G-S103A-V104I-A232V-N238Y-Q245R-A272V, T022K-S101N-S103N-V104I-A232V-Q245W, T022Q-S101T-S103N-V104I-A232M-Q245S, T022A-S078N-S101N-S103A-V104I-Q109G-L217Q-A232V-Q245R, W113Y, T022A-S101A-S103N-V104I-A232M-Q245S, T022A-S101Q-S103A-V104I-S128A-A232V-Q245R, G020R-S024R-S078G-G118D-G159D-S188D-H249R, T022A-S101N-S103G-V104I-A232T-Q245R, T022K-S101G-S103G-V104I-A232L-Q245R, T022A-N076D-G097A-S101Q-S103A-V104I-S128A-A232V-Q245R, T022Q-S101T-S103N-V104I-A232V-Q245S, T022A-S024F-S101G-S103A-V104I-N116L-Y209V-T213A-A232V, T022A-S101N-S103G-V104L-A232T-Q245R, T022A-S024R-S101G-S103A-V104I-P129E-A232V-N238Y-Q245R, T022A-N076D-G097S-S101Q-S103A-V104I-A232V-Q245R, T022A-N076D-S078N-G097A-S101N-S103A-V104I-A232V-Q245R, T022A-S101N-S103A-V104I-Q109G-S128A-A232V-Q245R, S259W, A016S-T022A-S024F-S101G-S103A-V104I-N116A-Y209V-G211Q-A232V, T022W-S101A-P210I-G211Q-T213A, T022A-S024G-N076D-G097S-S101N-S103A-V104I-A232V-Q245R, V028T-A215V, S024R-S078D-G118S-S188D-Q245R, T022Q-S101T-S103A-V104I-A232V-Q245W, T022A-S101N-S103A-V104I-Q109G-L217Q-A232V-Q245R, T022A-S024G-T071A-G097S-S101Q-S103A-V104I-S128A-A232V-Q245R, H120P, G020R-S024R-N062E-S078D-G118D-Q245R, T022A-N076D-S101Q-S103A-V104I-A232V-Q245R, T022Q-S101A-S103N-V104I-A232L-Q245W, T022Q-S101N-S103A-V104L-A232T-Q245R, T022W-S024R-N043R-R045T-S101A-N116L-P210I-G211Q, T022A-G097S-S101Q-S103A-V104I-S128A-L217Q-A232V-Q245R, G020R-S078G-G118S-G159D-S188D-H249R, A013S, A232N, T022A-N076D-S101G-S103A-V104I-A232V-Q245R, T022A-S024G-N076D-S078N-G097S-S101N-S103A-V104I-A232V-Q245R, I035T, V051M, N123A, G020R-S078G-G118S-G159D-Q245R-N248D, T022Y-S101A-S103N-V104I-A232L-Q245W, T022A-S024R-S101G-S103A-V104I-P129E-A232V-Q245R-A270V, G020R-S078D-G118S, T022R-S101N-S103N-V104L-A232M-Q245W, T022A-S024G-N076D-G097S-S101G-S103A-V104I-S128A-L217Q-A232V-Q245R, T022A-S101G-S103N-V104L-A232V-Q245R, T022A-N076D-G097A-S101Q-S103A-V104I-A232V-Q245R, T022A-S024R-N076D-S101G-S103A-V104I-I107V-P129E-A232V-Q245R, S024R-R045T-S101A-P210I-G211Q, T022A-S078N-S101N-S103A-V104I-Q109N-S128A-A232V-Q245R, T022A-S024G-S078N-G097A-S101N-S103A-V104I-Q109N-A232V-Q245R, G020R-S024R-S078G-G118D-P129E-G159D-H249R, G020R-S078G-G118D-P129E-S188D-Q245R-H249R, P014W, G020R-S024R-S078G-G118S-G159D-S188D-Q245R-N248D, T022R-S101G-S103N-V104I-A232M-Q245W, T022R-A098V-S101G-S103N-V104L-A232T-Q245R, T022A-S101T-S103A-V104I-A232V-Q245W, G020R-T022W-R045T-S101A-T213A, N043R-R045T-S101A, T022K-S101N-S103A-V104I-A232V-Q245R, T022K-S101T-S103N-V104I-A232L-Q245S, S056V, G020R-T022W-R045T-S101A-G211Q, G020R-S024R-S078D-G118D-Q245R-N248D, G020R-S024R-S078D-G118S-G159D-V244L-Q245R-N248D, T022A-G097A-S101Q-S103A-V104I-Q109G-S128A-A232V-Q245R, T022A-N076D-G097S-S101G-S103A-V104I-A232V-Q245R, G020R-T022W-R045T-S101A-N116L-P210I, T022Q-S101N-S103N-V104I-A232M-Q245S, T022Y-S101G-S103N-V104I-A232V-Q245W, A114V, S141C, T022A-N076D-S078N-S101N-S103A-V104I-A232V-Q245R, S056H, S078D-G118D-Q245R, S024R-N043R-R045T-S101A-T213A, T022Y-S101T-S103G-V104I-A232V-Q245S, T022A-S024G-G097S-S101Q-S103A-V104I-S128A-L217Q-A232V-Q245R, N269W, S078G-G118S-G159D-Q245R-N248D-H249R, T022W-S024R-R045T-S101A, T022A-S024R-S101G-S103A-V104I-P129E-A232V-N238Y-Q245R-A270V-A272V, T022A-S024R-S101G-S103A-V104I-P129E-A232V-Q245R, T022A-S024R-N076D-S101G-S103A-V104I-I107V-P129E-A232V-N238Y-Q245R-A270V-A272V, A122F, S024R-S078D-G118S-P129E-G159D-Q245R-H249R, T022K-S101T-S103G-V104I-A232V-Q245S, S009G-T022A-S078N-S101Q-S103A-V104I-Q109G-S128A-L217Q-A232V-Q245R, A098Q, T022A-S101N-S103A-V104L-A232L-Q245R, T022A-S101A-S103N-V104I-A232T-Q245W, T022A-S024G-N076D-G097A-S101N-S103A-V104I-A232V-Q245R, S024R-S078G-G118D-G159D-H249R, T022A-S024G-N076D-G097S-S101N-S103A-V104I-S128A-A232V-Q245R, T022A-S024G-N076D-S078N-S101Q-S103A-V104I-S128A-A232V-Q245R, T022A-S101Q-S103A-V104I-L124V-S128A-P129S-A232V-Q245R, T022R-S101N-S103A-V104I-A232M-W241R-Q245W, R045T-S101A-P210I-G211Q, G020R-S024R-S078G-G118S-P129E-Q245V-N248E, T022Q-S101T-S103N-V104I-A232L-Q245W, T022A-S024G-S078N-S101N-S103A-V104I-Q109G-L217Q-A232V-Q245R, T022A-S024G-G097A-S101N-S103A-V104I-L217Q-A223S-A232V-Q245R, T022K-S101T-S103A-V104I-A232T-Q245W, N043R-S101A-G211Q, T022A-S101G-S103A-V104I-Q109N-S128A-A232V-Q245R, T022A-G097A-S101G-S103A-V104I-S128A-L217Q-A232V-Q245R, G020R-S078G-G118S-P129E-H249R, S024R-S078G-G118S-G159D-S188D-Q245R, N043R-R045T-S101A-N204S-P210I-T213A, T022R-S101N-S103A-V104L-A232V-Q245S, T022R-S101N-S103G-V104I-A232V-Q245S, T022K-S101T-S103N-V104I-A232M-Q245S, T022A-N076D-S101N-S103A-V104I-L217Q-A232V-Q245R, T022A-S024G-N076D-S078N-S101N-S103A-V104I-Q109G-S128A-A232V-Q245R, G020R-T022W-R045T-S101A-P210I-T213A, T022A-N076D-G097A-S101G-S103A-V104I-L217Q-A232V-Q245R, T022W-S101A-P210I, T022A-G097A-S101N-S103A-V104I-L126I-P129S-A232V-Q245R, T022A-S078D-S101N-S103A-V104I-S128A-A232V-Q245R, Q236L, T022Y-S101T-S103N-V104I-A232V-Q245S, A200D, G020R-S078G-G118S-S188D-H249R, T022A-S101G-S103N-V104I-A232T-Q245W, T022A-S024G-N076D-G097A-S101N-S103A-V104I-S128A-A232V-Q245R, G157I, G020R-S024R-S078G-G118S-P129E-G159D-S188D-Q245R, T022Y-S101A-S103G-V104L-A232T-Q245R, S024R-S078G-G118D-H249R, T022Q-S101A-S103N-V104L-A232T-Q245S, T022Y-S101A-S103N-V104I-A232T-Q245W, T022A-S024G-S101N-S103A-V104I-Q109N-L217Q-A232V-Q245R, T022A-S101G-S103N-V104I-A232M-Q245S, T022A-S024G-N076D-S078N-S101G-S103A-V104I-S128A-A232V-Q245R, T022A-S101G-S103N-V104I-A232M-Q245W, I008N, T022A-S101N-S103G-V104I-A232M-Q245S, V026A, T022R-S101N-S103A-V104L-A232T-Q245W, T022A-S101A-S103N-V104I-A232L-Q245S, T022A-S101N-S103A-V104I-A232T-Q245R, N043R-R045T-S101A-P210I-G211Q-T213A, T022A-S024G-N076D-G097A-S101Q-S103A-V104I-S128A-A232V-Q245R, A098F-S099T-S101A, A098F-S099T-S101G-V104L, P014N, A098Q-S103G-V104I, S099T-S103A-V104I, S087W, S024R-N062E-S078G-G118S-H249R, S024R-N043R-R045T-S101A-G211Q, N043R-R045T-S101A-G211Q, G020R-S078G-G118S-N248D, T022Q-S101N-S103N-V104I-A232T-Q245W, S078D-G118S-P129E-Q245R-H249R, T022Q-S101T-S103G-V104I-A232T-Q245S, T022A-S024G-S101N-S103A-V104I-Q109N-S128A-A232V-Q245R, S024F-S101G-S103A-V104I-N116L-Y209A-T213A-A232V, S101A-N116L-P210I-T213A, T022R-S101N-S103N-V104L-A232L-Q245S, B015T, T022A-S101G-S103A-V104I-L124V-A232V-Q245R, G020R-S024R-S078D-G118D-G159D-Q245R, A114I, S212A, T022A-S101T-S103N-V104I-A232L-Q245S, T022A-S024G-S078N-G097S-S101N-S103A-V104I-S128A-T143I-L217Q-A232V-Q245R, S024R-S078G-G118S-G159D-S188D-Q245R-H249R, T022K-S101A-S103N-V104L-A232L-Q245S, T022W-S101A-T213A, T022A-N076D-S078N-G097S-S101Q-S103A-V104I-A232V-Q245R, S132V, S024R-S078G-G118D-S188D-Q245R-N248D-H249R, A098Q-S101A-S103A-V104L, A098Q-S099T-S103A, T022R-S101T-S103N-V104L-A232T-Q245W, T022A-N076D-S101Q-S103A-V104I-S128A-A232V-Q245R, V051A-S101A-P210I-T213A, A098Q-S099T-S101A-V104L, T022R-S101N-S103N-V104I-A232M-Q245W, S099A-S101A-S103A, G020R-S078G-G118D-S188D-Q245R, G020R-S024R-S078D-G118S-P129E-G159D-S188D-Q245R-H249R, T022A-N076D-S078N-G097A-S101Q-S103A-V104I-L217Q-A232V-Q245R, S024R-S078G-G118D, T022Q-S101N-S103N-V104L-A232M-Q245R, T022A-S101T-S103G-V104I-A232M-Q245S, A048G, T022Q-S099G-S101A-S103N-V104I-A232V-Q245R, S024R-R045T-S101A-G211Q-T213A, S024R-R045T-S101A-I107V-A114T-N116L-P210I-G211Q-T213A, T022R-S101T-S103G-V104I-A232L-Q245S, S099G-S103A, A098F-S099A, T022R-S101N-S103G-V104I-A232V-Q245W, T022A-S103A-V104I-L124T-A232V-Q245R, T022A-S024F-S101G-S103A-V104I-N116L-Y209V-A232V, T022K-S101N-S103N-V104L-A232V-Q245S, T022K-S101A-S103A-V104L-A232M-Q245R, T022Q-S101G-S103N-V104I-A232T-Q245W, T022A-S101T-S103A-V104L-A232T-Q245W, T022A-S101N-S103N-V104I-A232V-Q245S, T022A-N076D-S078N-G097A-S101N-S103A-V104I-L217Q-A232V-Q245R, A098M, T022A-S101G-S103G-V104I-A232L-Q245R, G020R-S101A-S207G-P210I-T213A, T022A-S024G-G097A-S101Q-S103A-V104I-Q109G-S128A-A232V-Q245R, G020R-S024R-S078G-G118S-G159D-Q245R-N248D-H249R, G020R-S024R-S078D-G118S-P129E-G159D-Q245R, T022Y-S101G-S103N-V104I-A232L-Q245S, V026Q, S240G, G020R-S024R-S078G-G118S-S188D, G020R-S024R-S078G-G118D-N238D-N248D, T022R-S101G-S103A-V104I-A232T-Q245S, T022W-S024R-R045T-S101A-P210I-G211Q, T022A-S024G-N076D-S078N-G097S-S101Q-S103A-V104I-A232V-Q245R, T022W-S024R-R045T-S101A-T213A, A200F, T022R-S101T-S103G-V104L-A232T-Q245S, T022A-S078N-G097S-S101Q-S103A-V104I-Q109N-L217Q-A232V-Q245R, T022A-S101G-S103N-V104I-A232T-Q245S, T022A-S101T-S103N-V104I-A232T-Q245S, T022A-S101A-S103A-V104L-A232V-Q245S, S099G-S101A-V104L, H017R-T022A-S024V-N076D-G097S-S101N-S103A-V104I-Q109N-S128A-A232V-Q245R, T022A-S101A-S103N-V104L-A232M-Q245S, T022A-S101Q-S103A-V104I-Q109G-L217Q-A232V-Q245R, T022A-S024G-N076D-S101Q-S103A-V104I-S128A-A232V-Q245R, T022A-S024G-S078N-G097S-S101N-S103A-V104I-S128A-S212P-L217Q-A232V-Q245R, S056L, T253L, T022W-R045T-S101A-G211Q-T213A, T022A-G097A-S101Q-S103A-V104I-S128A-L217Q-A232V-Q245R, T022A-S024G-N076D-S078N-G097A-S101N-S103A-V104I-S128A-A232V-Q245R, G020R-T022W-R045T-S101A-G211Q-T213A, S101A-G211Q, N123D, T022R-S101A-S103N-V104I-A232M-Q245W, T022Y-S101T-S103N-V104L-A232T-Q245R, T022A-S101G-S103A-V104I-N116L-Y209V-G211Q-T213A-A232V, G020R-S024R-S078D-G118D-N248D, A098Q-S099G-S101A, S099T-V104I, T022Q-S101G-S103A-V104I-A232V-Q245S, T022R-S101A-S103G-V104I-A232T-Q245S, T022K-S101T-S103N-V104I-A232T-Q245W, A016M, T022A-S024G-N076D-S101N-S103A-V104I-Q109G-A232V-Q245R, T022K-S101N-S103N-V104I-A232T-Q245S, G020R-N043R-R045T-S101A-N204D-P210I-G211Q, T022A-S101T-S103N-V104I-A232V-Q245S, T022Y-S101G-S103N-V104L-A232V-Q245R, T022A-S101T-S103N-V104I-A232V-Q245W, T022A-S024G-S078N-G097S-S101N-S103A-V104I-Q109G-L217Q-A232V-Q245R, A088T, T022Y-S101A-S103A-V104I-A232M-Q245S, T022W-S024R-R045T-S101A-N116L-P210I-G211Q, T022W-N043R-R045T-S101A-P210I-G211Q-T213A, T038A, S078D-G118S-G159D-Q245R, T022K-S101T-S103A-V104L-A232V-Q245W, T022A-S024G-N076D-S078N-G097S-S101N-S103A-V104I-Q109G-A232V-Q245R, T022A-S078N-S101Q-S103A-V104I-Q109N-L217Q-A232V-Q245R, G025S, G025T, A016S-S101G-S103A-V104I-N116L-M119V-Y209V-G211Q-A232V, T022A-N076D-V084A-S101G-S103A-V104I-A232V-Q245R, V011L, T022Y-S101T-S103G-V104I-A232M-Q245S, T022R-S101N-S103A-V104L-A232T-Q245S, S240V, G020R-S024R-S078G-G118D-P129E-S188D-H249R, T022W-S024R-R045T-S101A-G211Q-T213A, T022A-S024G-N076D-G097A-S101Q-S103A-V104I-Q109G-A232V-Q245R, V004Y, V051A, G195W-N269E, S024R-S078G-G118D-N248D-H249R, T022A-S099G-S101T-S103G-V104I-A232V-Q245S, T022Q-S101T-S103G-V104I-A232L-Q245W, T022Q-S101N-S103N-V104I-A232V-Q245W, T022A-S101T-S103G-V104I-A232V-Q245W, T022K-S101N-S103G-V104I-A232V-Q245R, T022Q-S101G-S103N-V104I-A232V-Q245S, T022R-S078N-G097S-S101G-S103A-V104I-Q109G-S128A-A232V-Q245R, T022R-S101G-S103A-V104I-M222S-A232V-Q245R-E271H, G020R-S024R-S101N-Q109G-N116L-M222S, R010A-G020R-S024R-S101A-N116L или G020R-N043R-M222S, где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
17. Выделенный вариант субтилизина по любому из пп. 1-16, где вариант субтилизина представляет собой вариант субтилизина из протеазы субтилизин GG36 Bacillus lentus, где указанная протеаза субтилизин GG36 Bacillus lentus содержит аминокислотную последовательность, приведенную в SEQ ID №:2.
18. Выделенный вариант субтилизина по любому из пп. 1-16, где указанный вариант субтилизина имеет по меньшей мере 90% идентичности с указанной протеазой субтилизин GG36 Bacillus lentus, содержащей аминокислотную последовательность, приведенную в SEQ ID №:2.
19. Выделенный вариант субтилизина по любому из пп. 1-16, где общий заряд варианта составляет 0, +1, +2, +3, +4, +5, -1, -2, -3, -4 или -5 по отношению к общему заряду протеазы субтилизин GG36 Bacillus lentus.
20. Выделенный вариант субтилизина по п. 19, где общий заряд варианта составляет -1, 0, +1, +2, +3, +4 или +5 по отношению к общему заряду протеазы субтилизин GG36 Bacillus lentus.
21. Выделенный вариант субтилизина по п. 20, где общий заряд варианта составляет -1, 0, +1, +2, +3 или +4 по отношению к общему заряду протеазы субтилизин GG36 Bacillus lentus.
22. Выделенный вариант субтилизина по п. 19, где общий заряд варианта составляет +1, 0, -1, -2, -3, -4 или -5 по отношению к общему заряду протеазы субтилизин GG36 Bacillus lentus.
23. Выделенный вариант субтилизина по п. 22, где общий заряд варианта составляет +1, 0, -1, -2, -3 или -4 по отношению к общему заряду протеазы субтилизин GG36 Bacillus lentus.
24. Выделенный вариант субтилизина по любому из пп. 1-16, где общий заряд получен с помощью одной или нескольких замен, выбранных из: A001E, V004E, R010H/A, Q012E, A015D, N018D, R019H/S, V026D, K027E, N043D, R045C/T/S/P, S049D, V051E, G061E, N062D/E, N076D, N077D, S078D, S087D, A098E, S101D, S106E, G115E, G118D, N123D, S128D, P129E, S130D/E, S132D, A158E, G159D/E, S160D, S166D, R170T, N183D, N184D, N185E, R186H, S188D/E, A194E, A200D, Y209E, A215D, N204D, S212D, L217D/E, N218D, A230E, K235F, N237D, K237E, N238D, S240D, N243D, Q245D, R247L, N248D/E, K251C, N263D, N269D/E, A272D, A273E или R275H/S, и где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
25. Выделенный вариант субтилизина по любому из пп. 1-16, где общий заряд получен с помощью одной или нескольких замен, выбранных из: A001R, V004R, Q012R, P014R, H017R, N018R/K, G020K/R, T022R/K, S024R/K, G025R, D032I, D041K/L/N, N043R/K, G046R, A048R, F050R/K, P055R, S056R/K, T057R, Q059K, G061R, N076K, S078R, P086R, S087R, E089G/P/I, G097R, S099R, Q109R, G115R, G118R, S132K, S144R, G159R/K, D181C/S/T, L196K, N204K, Q206R, K235R, Q236R/K, K237R, N238R, S240R, W241R, S242R/K, V244R, Q245R/K, N248R, H249R, N252R/K, S256R, G258R, T260K, N269R/K или E271F/H/T/L/W/R/S/I/A/G/V, и где положения аминокислот варианта субтилизина пронумерованы согласно нумерации соответствующих положений аминокислот в аминокислотной последовательности субтилизина BPN′ Bacillus amyloliquefaciens, приведенной в SEQ ID №:1.
26. Выделенный вариант субтилизина по любому из пп. 1-16, где вариант субтилизина имеет одну или несколько из следующих характеристик: a) эффективность по методике испытания 2, равную по меньшей мере 1,1, по меньшей мере 1,2, по меньшей мере 1,3, по меньшей мере 1,4, по меньшей мере 1,5, по меньшей мере 1,6, по меньшей мере 1,7, по меньшей мере 1,8, по меньшей мере 1,9, по меньшей мере 2, от 1,1 до примерно 10, от 1,1 до примерно 8 или даже от 1,1 до примерно 5; b) эффективность по методике испытания 3, равную по меньшей мере 1,1, по меньшей мере 1,2, по меньшей мере 1,3, по меньшей мере 1,4, по меньшей мере 1,5, по меньшей мере 1,6, по меньшей мере 1,7, по меньшей мере 1,8, по меньшей мере 1,9, по меньшей мере 2, от 1,1 до примерно 10, от 1,1 до примерно 8 или даже от 1,1 до примерно 5; c) эффективность по методике испытания 4, равную по меньшей мере 1,0, по меньшей мере 1,1, по меньшей мере 1,2, по меньшей мере 1,3, по меньшей мере 1,4, по меньшей мере 1,5, по меньшей мере 1,6, по меньшей мере 1,7, по меньшей мере 1,8, по меньшей мере 1,9, по меньшей мере 2, от 1,0 до примерно 10, от 1,0 до примерно 8 или даже от 1,0 до примерно 5; и/или d) эффективность по методике испытания 6, равную по меньшей мере 1,0, равную по меньшей мере 1,1, по меньшей мере 1,2, по меньшей мере 1,3, по меньшей мере 1,4, по меньшей мере 1,5, по меньшей мере 1,6, по меньшей мере 1,7, по меньшей мере 1,8, по меньшей мере 1,9, по меньшей мере 2, от 1,0 до примерно 10, от 1,0 до примерно 8 или даже от 1,0 до примерно 5; и/или е) эффективность по методике испытания 7, равную по меньшей мере 1,1, по меньшей мере 1,2, по меньшей мере 1,3, по меньшей мере 1,4, по меньшей мере 1,5, по меньшей мере 1,6, по меньшей мере 1,7, по меньшей мере 1,8, по меньшей мере 1,9, по меньшей мере 2, от 1,1 до примерно 15, от 1,1 до примерно 10 или даже от 1,1 до примерно 7.
27. Выделенная нуклеиновая кислота, содержащая полинуклеотидную последовательность, которая кодирует вариант субтилизина, представленный в пп. 1-26.
28. Вектор экспрессии, содержащий нуклеиновую кислоту по п. 27.
29. Вектор экспрессии по п. 28, где указанная по меньшей мере одна нуклеиновая кислота функционально связана с промотором.
30. Клетка-хозяин, включающая вектор экспрессии по любому из пп. 28 или 29.
31. Способ получения по меньшей мере одного варианта субтилизина из субтилизина Bacillus, включающий:
a) трансформацию клетки-хозяина вектором экспрессии, включающим по меньшей мере одну нуклеиновую кислоту, кодирующую по меньшей мере один вариант субтилизина по любому из пп. 1-26, с получением трансформированной клетки-хозяина; и
b) культивирование трансформированной клетки-хозяина в условиях, подходящих для получения по меньшей мере одного варианта субтилизина, с получением по меньшей мере одного варианта субтилизина.
32. Способ по п. 31, дополнительно включающий сбор полученного варианта субтилизина.
33. Способ по любому из пп. 30-31, где указанная клетка-хозяин принадлежит к виду рода Bacillus.
34. Способ по п. 33, где указанным видом рода Bacillus является B. subtilis.
35. Композиция, включающая по меньшей мере один вариант субтилизина по любому из пп. 1-26, где указанная композиция не является продуктом по уходу за тканью и по уходу за домом.
36. Композиция по п. 35, где указанная композиция представляет собой чистящую композицию.
37. Композиция по п. 35, где указанная чистящая композиция является гранулированной, порошковой, твердой, брусковой, жидкой, таблетированной, гелеобразной или пастообразной композицией.
38. Композиция по п. 35, дополнительно включающая по меньшей мере одно отбеливающее средство.
39. Композиция по п. 35, где указанная чистящая композиция является бесфосфатной.
40. Композиция по любому из пп. 35, где указанная чистящая композиция содержит фосфат.
41. Композиция по п. 35, дополнительно включающая по меньшей мере один дополнительный фермент.
42. Композиция по п. 41, где указанный по меньшей мере один дополнительный фермент выбран из гемицеллюлаз, целлюлаз, пероксидаз, протеаз, металлопротеаз, ксиланаз, липаз, фосфолипаз, эстераз, пергидролаз, кутиназ, пектиназ, пектатлиаз, маннаназ, кератиназ, редуктаз, оксидаз, фенолоксидаз, липоксигеназ, лигниназ, пуллуланаз, танназ, пентозаназ, маланаз, β-глюканаз, арабинозидаз, гиалуронидаз, хондроитиназ, лакказ и амилаз или любой их комбинации.
43. Композиция по п. 35, где указанный вариант субтилизина не является протеазой для холодной воды.
44. Способ чистки, включающий приведение в контакт поверхности или предмета с чистящей композицией, включающей по меньшей мере один вариант субтилизина по любому из пп. 1-26.
45. Способ чистки, включающий приведение в контакт поверхности или предмета с чистящей композицией, изложенной в любом из пп. 35-43.
46. Способ по любому из пп. 44-45, дополнительно включающий ополаскивание указанной поверхности или предмета после приведения в контакт указанной поверхности или предмета, соответственно, с указанной чистящей композицией.
47. Способ по любому из пп. 44-45, дополнительно включающий стадию ополаскивания указанной поверхности или предмета после приведения в контакт указанной поверхности или предмета с указанной чистящей композицией.
48. Способ по п. 47, дополнительно включающий стадию сушки указанной поверхности или предмета после указанного ополаскивания указанной поверхности или предмета.
49. Способ чистки поверхности или предмета, включающий: обеспечение чистящей композиции по любому из пп. 35-43 и поверхности или предмета, нуждающихся в чистке; и приведение указанной чистящей композиции в контакт с указанной поверхностью или предметом, нуждающимися в чистке, в условиях, подходящих для чистки указанной поверхности или предмета, с получением очищенной поверхности или предмета.
50. Способ по п. 49, дополнительно включающий стадию ополаскивания указанной очищенной поверхности или с получением ополоснутой поверхности или предмета.
51. Способ по любому из пп. 49-50, дополнительно включающий стадию сушки указанной ополоснутой поверхности или предмета.
52. Способ по любому из пп. 44, 45 или 49, где указанный вариант субтилизина не является вариантом протеазы для холодной воды.
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CA2834865A1 (en) | 2012-11-08 |
JP6522026B2 (ja) | 2019-05-29 |
WO2012151534A1 (en) | 2012-11-08 |
EP2705146A1 (en) | 2014-03-12 |
EP3486319A2 (en) | 2019-05-22 |
CA3111256A1 (en) | 2012-11-08 |
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AR086281A1 (es) | 2013-12-04 |
ES2707869T3 (es) | 2019-04-05 |
EP2705146B1 (en) | 2018-11-07 |
JP2017184726A (ja) | 2017-10-12 |
MX2013012690A (es) | 2014-03-21 |
RU2718648C2 (ru) | 2020-04-10 |
US9856466B2 (en) | 2018-01-02 |
CN103764823B (zh) | 2018-05-11 |
TR201901382T4 (tr) | 2019-02-21 |
JP2014522235A (ja) | 2014-09-04 |
MX338925B (es) | 2016-05-06 |
JP6105560B2 (ja) | 2017-03-29 |
EP4230735A1 (en) | 2023-08-23 |
CA3111256C (en) | 2023-05-09 |
US20140154782A1 (en) | 2014-06-05 |
EP3486319A3 (en) | 2019-07-10 |
CA3194062A1 (en) | 2012-11-08 |
CA2834865C (en) | 2021-03-09 |
EP3486319B1 (en) | 2022-11-30 |
CN103764823A (zh) | 2014-04-30 |
BR112013027963A2 (pt) | 2016-11-29 |
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