WO2001042432A3 - Modified enzymatic activity through subdomain swaps between related alpha/beta-barrel enzymes - Google Patents

Modified enzymatic activity through subdomain swaps between related alpha/beta-barrel enzymes Download PDF

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Publication number
WO2001042432A3
WO2001042432A3 PCT/GB2000/004661 GB0004661W WO0142432A3 WO 2001042432 A3 WO2001042432 A3 WO 2001042432A3 GB 0004661 W GB0004661 W GB 0004661W WO 0142432 A3 WO0142432 A3 WO 0142432A3
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WO
WIPO (PCT)
Prior art keywords
enzymes
subdomain
swaps
beta
substrate
Prior art date
Application number
PCT/GB2000/004661
Other languages
French (fr)
Other versions
WO2001042432A2 (en
Inventor
Alan Fersht
Myriam Altamirano
Original Assignee
Medical Res Council
Alan Fersht
Myriam Altamirano
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Medical Res Council, Alan Fersht, Myriam Altamirano filed Critical Medical Res Council
Priority to AU17205/01A priority Critical patent/AU1720501A/en
Publication of WO2001042432A2 publication Critical patent/WO2001042432A2/en
Publication of WO2001042432A3 publication Critical patent/WO2001042432A3/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

Methods of obtaining enzymes that bind target substrate and catalyse desired reactions. α/β-barrel proteins are categorised into two classes based on catalytic lid structure. Lids can be grafted onto scaffolds with additional minor modifications at conserved and non-conserved residues to provide candidate product enzymes for screening for the desired properties. Design of a novel enzyme which binds a target substrate and catalyses a reaction of choice is facilitated by selection of a scaffold which binds the substrate and of a catalytic lid of the correct class for the desired reaction. Targeted or focussed mutagenesis may be used to refine substrate binding and catalysis.
PCT/GB2000/004661 1999-12-08 2000-12-06 Modified enzymatic activity through subdomain swaps between related alpha/beta-barrel enzymes WO2001042432A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU17205/01A AU1720501A (en) 1999-12-08 2000-12-06 Methods of producing novel enzymes

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GBGB9929061.1A GB9929061D0 (en) 1999-12-08 1999-12-08 Methods of producing novel enzymes
GB9929061.1 1999-12-08

Publications (2)

Publication Number Publication Date
WO2001042432A2 WO2001042432A2 (en) 2001-06-14
WO2001042432A3 true WO2001042432A3 (en) 2002-05-10

Family

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Family Applications (1)

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PCT/GB2000/004661 WO2001042432A2 (en) 1999-12-08 2000-12-06 Modified enzymatic activity through subdomain swaps between related alpha/beta-barrel enzymes

Country Status (3)

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AU (1) AU1720501A (en)
GB (2) GB9929061D0 (en)
WO (1) WO2001042432A2 (en)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2003523742A (en) * 2000-02-03 2003-08-12 ドマンティス リミテッド Combinatorial protein domains
AU2001277614A1 (en) * 2000-08-07 2002-02-18 Domantis Limited Novel proteins
EP1456362A2 (en) * 2001-02-09 2004-09-15 California Institute of Technology Method for the generation of proteins with new enzymatic function
US7335504B2 (en) * 2003-06-18 2008-02-26 Direvo Biotechnology Ag Engineered enzymes and uses thereof
AU2006304804B2 (en) 2005-10-21 2011-06-02 Vertex Pharmaceuticals Incorporated Modified proteases that inhibit complement activation
US9493747B2 (en) 2008-01-03 2016-11-15 The General Hospital Corporation Engineered transglutaminase barrel proteins

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994005772A1 (en) * 1992-09-08 1994-03-17 Rutgers, The State University Of New Jersey Improved enzymes for the production of 2-keto-l-gulonic acid

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994005772A1 (en) * 1992-09-08 1994-03-17 Rutgers, The State University Of New Jersey Improved enzymes for the production of 2-keto-l-gulonic acid

Non-Patent Citations (9)

* Cited by examiner, † Cited by third party
Title
ALTAMIRANO MYRIAM M ET AL: "Directed evolution of new catalytic activity using the alpha/beta-barrel scaffold.", NATURE (LONDON), vol. 403, no. 6770, 10 February 2000 (2000-02-10), pages 617 - 622, XP002173865, ISSN: 0028-0836 *
CHEN Z ET AL: "COMPLEMENTING AMINO ACID SUBSTITUTIONS WITHIN LOOP 6 OF THE ALPHA-BETA-BARREL ACTIVE SITE INFLUENCE THE CARBON DIOXIDE OXYGEN SPECIFICITY OF CHLOROPLAST RIBULOSE-1 5-BISPHOSPHATE CARBOXYLASE-OXYGENASE", BIOCHEMISTRY, vol. 30, no. 36, 1991, pages 8846 - 8850, XP002173864, ISSN: 0006-2960 *
DARIMONT BEATRICE ET AL: "Mutational analysis of the active site of indoleglycerol phosphate synthase from Escherichia coli.", PROTEIN SCIENCE, vol. 7, no. 5, May 1998 (1998-05-01), pages 1221 - 1232, XP000993562, ISSN: 0961-8368 *
FERSHT ALAN R ET AL: "Designing new enzymes.", BIOCHEMICAL SOCIETY TRANSACTIONS, vol. 28, no. 3, 2000, 671st Meeting of the Biochemical Society.;England, UK; April 11-13, 2000, pages A53, XP000993578, ISSN: 0300-5127 *
HOPFNER K P ET AL: "New enzyme lineages by subdomain shuffling", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF USA,NATIONAL ACADEMY OF SCIENCE. WASHINGTON,US, vol. 95, no. 17, August 1998 (1998-08-01), pages 9813 - 9818, XP002103693, ISSN: 0027-8424 *
O'BRIEN PATRICK J ET AL: "Catalytic promiscuity and the evolution of new enzymatic activities.", CHEMISTRY & BIOLOGY (LONDON), vol. 6, no. 4, April 1999 (1999-04-01), pages R91 - R105, XP000993553, ISSN: 1074-5521 *
PUJADAS GERARD ET AL: "TIM barrel fold: Structural, functional and evolutionary characteristics in natural and designed molecules.", BIOLOGIA (BRATISLAVA), vol. 54, no. 3, June 1999 (1999-06-01), pages 231 - 254, XP000993511, ISSN: 0006-3088 *
STEHLIN CATHERINE ET AL: "Deletion mutagenesis as a test of evolutionary relatedness of indoleglycerol phosphate synthase with other TIM barrel enzymes.", FEBS LETTERS, vol. 403, no. 3, 1997, pages 268 - 272, XP002173863, ISSN: 0014-5793 *
WILMANNS M ET AL: "THREE-DIMENSIONAL STRUCTURE OF THE BIFUNCTIONAL ENZYME PHOSPHORIBOSYLANTHRANILATE ISOMERASE INDOLEGLYCEROLPHOSPHATE SYNTHASE FROM ESCHERICHIA-COLI REFINED AT 2.0 A RESOLUTION", JOURNAL OF MOLECULAR BIOLOGY, vol. 223, no. 2, 1992, pages 477 - 508, XP000993512, ISSN: 0022-2836 *

Also Published As

Publication number Publication date
GB0029781D0 (en) 2001-01-17
AU1720501A (en) 2001-06-18
WO2001042432A2 (en) 2001-06-14
GB2358633A (en) 2001-08-01
GB9929061D0 (en) 2000-02-02

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