JP6200578B2 - サイコースエピマー化酵素変異体及びこれを用いたサイコースの製造方法 - Google Patents
サイコースエピマー化酵素変異体及びこれを用いたサイコースの製造方法 Download PDFInfo
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Description
本願に記載されているエピマー化酵素変異体或いは前記変異体を生産するように組換えられた微生物を固定化させた担体で充填したカラムを含む、サイコース生産用固定化反応器に関する。
前記酵素反応物を精製することによってサイコースを収得する段階;を含んでもよい。
(1)置換させるアミノ酸配列位置の選定
Claims (13)
- アグロバクテリウム・ツメファシエンス(Agrobacterium tumefaciens)ATCC33970から由来したアミノ酸配列を有する野生型サイコースエピマー化酵素の変異体として、前記アミノ酸配列において77番目のアミノ酸であるグルタミン酸がプロリンに置換されたアミノ酸配列を有する、熱安定性が向上されたサイコースエピマー化酵素変異体。
- アグロバクテリウム・ツメファシエンス(Agrobacterium tumefaciens)ATCC33970から由来したアミノ酸配列を有する野生型サイコースエピマー化酵素の変異体として、前記アミノ酸配列において77番目のアミノ酸であるグルタミン酸がプロリンに置換され、前記アミノ酸配列において33番目のアミノ酸であるイソロイシンが、ロイシン及びシステインからなる群より選ばれた一つのアミノ酸に置換されるか、前記アミノ酸配列において213番目のアミノ酸であるセリンがシステインに置換されたアミノ酸配列を有する、熱安定性が向上されたサイコースエピマー化酵素変異体。
- アグロバクテリウム・ツメファシエンス(Agrobacterium tumefaciens)ATCC33970から由来したアミノ酸配列を有する野生型サイコースエピマー化酵素の変異体として、前記アミノ酸配列において77番目のアミノ酸であるグルタミン酸がプロリンに置換され、前記アミノ酸配列においてさらに33番目のアミノ酸であるイソロイシンがロイシンに置換され、前記アミノ酸配列においてさらに213番目のアミノ酸であるセリンがシステインに置換されたアミノ酸配列を有する、熱安定性が向上されたサイコースエピマー化酵素変異体。
- 請求項1〜請求項3のいずれか1項によるサイコースエピマー化酵素変異体をコーディングする、ポリヌクレオチド。
- 請求項1〜請求項3のいずれか1項によるサイコースエピマー化酵素変異体をコーディングする遺伝子を含む、組換えベクター。
- 請求項1〜請求項3のいずれか1項によるサイコースエピマー化酵素変異体を生産するように組換えられた、微生物。
- 前記微生物がコリネバクテリウム・グルタミカム(Corynebacterium glutamicum)である、請求項6に記載の微生物。
- 果糖に、請求項1〜請求項3のいずれか1項によるサイコースエピマー化酵素変異体、又は前記サイコースエピマー化酵素変異体を生産するように組換えされた微生物を提供し、酵素反応を起こす段階;及び
前記酵素反応物を精製することによってサイコースを収得する段階;を含む、サイコースの製造方法。 - 前記酵素反応段階で金属イオンがさらに提供される、請求項8に記載の製造方法。
- 前記金属がマンガンである、請求項9に記載の製造方法。
- 前記金属イオンが0.01mM〜5mMの濃度で追加される、請求項9に記載の製造方法。
- 請求項1〜請求項3のいずれか1項によるサイコースエピマー化酵素変異体又は請求項6による微生物を固定化させた担体で充填したカラムを含む、サイコース生産用固定化反応器。
- 請求項12による固定化反応器に果糖溶液を供給することによってサイコースを生産する方法。
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PCT/KR2014/003545 WO2014175655A1 (ko) | 2013-04-23 | 2014-04-23 | 사이코스 에피머화 효소 변이체 및 이를 이용한 사이코스의 제조 방법 |
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KR101789837B1 (ko) | 2009-09-17 | 2017-10-25 | 아이아이아이 홀딩스 3, 엘엘씨 | 자기저항소자 및 이를 이용한 비휘발성 반도체 기억장치 |
KR102087396B1 (ko) * | 2015-11-16 | 2020-03-10 | 주식회사 삼양사 | 과당-함유 기질로부터 사이코스를 생산하는 방법 |
ES2861065T3 (es) | 2015-12-23 | 2021-10-05 | Cj Cheiljedang Corp | Composición para producir D-psicosa que comprende D-psicosa 3-epimerasa y sal y método para producir D-psicosa utilizando la misma |
KR101723007B1 (ko) | 2016-02-29 | 2017-04-04 | 씨제이제일제당(주) | 고순도 d-사이코스를 제조하는 방법 |
CN106148311B (zh) * | 2016-09-12 | 2019-06-21 | 上海立足生物科技有限公司 | 一种d-阿洛酮糖-3-差向异构酶的突变体及其应用 |
CN110462036B (zh) * | 2016-11-16 | 2023-07-14 | Cj第一制糖株式会社 | 一种新型d-阿洛酮糖3-差向异构酶以及使用该酶制备d-阿洛酮糖的方法 |
MA46152B1 (fr) | 2016-12-14 | 2020-11-30 | Bonumose Llc | Production enzymatique de d-allulose |
WO2018116266A1 (en) * | 2016-12-23 | 2018-06-28 | Petiva Private Ltd. | D-psicose 3-epimerase mutant and uses thereof |
KR101981430B1 (ko) | 2017-06-23 | 2019-05-23 | 씨제이제일제당 (주) | D-사이코스 붕산염 착물로부터 크로마토그래피를 이용한 d-사이코스의 생산 방법 및 d-사이코스를 포함하는 조성물 |
KR102114865B1 (ko) * | 2017-12-08 | 2020-05-27 | 씨제이제일제당 주식회사 | 신규한 사이코스-6-인산 탈인산효소, 상기 효소를 포함하는 사이코스 생산용 조성물, 상기 효소를 이용하여 사이코스를 제조하는 방법 |
US11104893B2 (en) | 2018-01-24 | 2021-08-31 | Matsutani Chemical Industry Co., Ltd. | Ketose 3-epimerase with improved thermal stability |
KR102439295B1 (ko) | 2018-11-30 | 2022-09-02 | 씨제이제일제당 주식회사 | D-사이코스 결정 및 이의 제조 방법 |
KR102138862B1 (ko) * | 2019-03-08 | 2020-07-30 | 씨제이제일제당 주식회사 | 알룰로스를 생산하는 스태필로코커스 속 미생물 및 이를 이용한 알룰로스 제조방법 |
KR102487975B1 (ko) * | 2019-10-31 | 2023-01-12 | 주식회사 삼양사 | 우수한 전환 활성을 갖는 균체 고정화 비드 및 이의 제조방법 |
KR102448351B1 (ko) * | 2020-04-27 | 2022-09-28 | 대상 주식회사 | 알룰로스 에피머화 효소 변이체, 이의 제조방법 및 이를 이용한 알룰로스의 제조방법 |
JP2024505616A (ja) | 2021-12-29 | 2024-02-07 | テサン・コーポレイション | 常時発現用新規プロモーター変異体およびその用途 |
CN114958815B (zh) * | 2022-04-29 | 2023-07-25 | 西北工业大学 | 一种d-阿洛酮糖3-差向异构酶及其固定化方法 |
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