Title:
MESO-2,3-BUTANEDIOL DEHYDROGENASE, AND MUTANT AND USE THEREOF
Document Type and Number:
WIPO Patent Application WO/2023/155474
Kind Code:
A1
Abstract:
Disclosed are a meso-2,3-butanediol dehydrogenase and a mutant and use thereof. The meso-2,3-butanediol dehydrogenase has an amino acid sequence set forth in SEQ ID NO: 1. According to the present invention, a meso-2,3-butanediol dehydrogenase with high-temperature resistance is found; on the basis of the meso-2,3-butanediol dehydrogenase, by using an accelerated sampling molecular dynamics simulation method to dynamically describe the release process of a product, the problem that high stability and high activity cannot be balanced is solved, and a mutant capable of catalyzing the preparation of meso-2,3-butanediol is obtained, which has very good high-temperature resistance. After heat treatment at 100 °C for 30 min, the residual enzyme activity is 23.9%. In catalyzing the formation of meso-2,3-butanediol by acetoin, the activity is increased by a factor of about 2-5. The product meso-2,3-butanediol obtained for the reaction has extremely high optical purity, which provides a broad application prospect for the production of meso-2,3-butanediol by bioconversion.
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Inventors:
YU HAORAN (CN)
PU ZHONGJI (CN)
PU ZHONGJI (CN)
Application Number:
PCT/CN2022/127954
Publication Date:
August 24, 2023
Filing Date:
October 27, 2022
Export Citation:
Assignee:
ZJU HANGZHOU GLOBAL SCIENTIFIC AND TECH INNOVATION CENTER (CN)
International Classes:
C12N9/04; C12N1/21; C12N15/53; C12N15/70; C12P7/18; C12R1/19
Foreign References:
| CN114703156A | 2022-07-05 | |||
| CN108368523A | 2018-08-03 | |||
| CN105200068A | 2015-12-30 | |||
| US20050003500A1 | 2005-01-06 |
Other References:
蒲中机 (PU, ZHONGJI): "基于酶催化循环理性设计NAD+依赖的氧化还原酶 (Non-official translation: Rational Design of NAD+-dependent Oxidoreductases Based on Enzyme-catalyzed Cycles)", 中国博士学位论文全文数据库 (CHINA DOCTORAL DISSERTATIONS FULL-TEXT DATABASE), no. 3, 15 March 2022 (2022-03-15), ISSN: 1674-022X
DATABASE Protein 7 March 2021 (2021-03-07), ANONYMOUS : "MULTISPECIES: (S)-acetoin forming diacetyl reductase [Lactococcus] ", XP093086331, retrieved from NCBI Database accession no. WP_011835462.1
RADOš DUšICA; TURNER DAVID L.; CATARINO TERESA; HOFFART EUGENIA; NEVES ANA RUTE; EIKMANNS BERNHARD J.; BLOMBACH BASTIAN;: "Stereospecificity ofCorynebacterium glutamicum2,3-butanediol dehydrogenase and implications for the stereochemical purity of bioproduced 2,3-butanediol", APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, SPRINGER BERLIN HEIDELBERG, BERLIN/HEIDELBERG, vol. 100, no. 24, 29 September 2016 (2016-09-29), Berlin/Heidelberg, pages 10573 - 10583, XP036105347, ISSN: 0175-7598, DOI: 10.1007/s00253-016-7860-6
HAO WENBO, JI FANG-LING, WANG JING-YUN, ZHANG YUE, WANG TIAN-QI, CHE WEN-SHI, BAO YONG-MING: "Effects of DI 94G Mutant on meso-2, 3-Butanediol Dehydrogenase Catalytic Properties", ZHONGGUO SHENGWU GONGCHENG ZAZHI - CHINA BIOTECHNOLOGY, ZHONGGUO KEXUEYUAN WENXIAN QINGBAO ZHONGXIN,CHINESE ACADEMY OF SCIENCES, DOCUMENTATION AND INFORMATION CENTER, CN, vol. 36, no. 1, 11 January 2016 (2016-01-11), CN, pages 47 - 54, XP093086338, ISSN: 1671-8135, DOI: 10.13523/j.cb.20160107
DATABASE Protein 7 March 2021 (2021-03-07), ANONYMOUS : "MULTISPECIES: (S)-acetoin forming diacetyl reductase [Lactococcus] ", XP093086331, retrieved from NCBI Database accession no. WP_011835462.1
RADOš DUšICA; TURNER DAVID L.; CATARINO TERESA; HOFFART EUGENIA; NEVES ANA RUTE; EIKMANNS BERNHARD J.; BLOMBACH BASTIAN;: "Stereospecificity ofCorynebacterium glutamicum2,3-butanediol dehydrogenase and implications for the stereochemical purity of bioproduced 2,3-butanediol", APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, SPRINGER BERLIN HEIDELBERG, BERLIN/HEIDELBERG, vol. 100, no. 24, 29 September 2016 (2016-09-29), Berlin/Heidelberg, pages 10573 - 10583, XP036105347, ISSN: 0175-7598, DOI: 10.1007/s00253-016-7860-6
HAO WENBO, JI FANG-LING, WANG JING-YUN, ZHANG YUE, WANG TIAN-QI, CHE WEN-SHI, BAO YONG-MING: "Effects of DI 94G Mutant on meso-2, 3-Butanediol Dehydrogenase Catalytic Properties", ZHONGGUO SHENGWU GONGCHENG ZAZHI - CHINA BIOTECHNOLOGY, ZHONGGUO KEXUEYUAN WENXIAN QINGBAO ZHONGXIN,CHINESE ACADEMY OF SCIENCES, DOCUMENTATION AND INFORMATION CENTER, CN, vol. 36, no. 1, 11 January 2016 (2016-01-11), CN, pages 47 - 54, XP093086338, ISSN: 1671-8135, DOI: 10.13523/j.cb.20160107
Attorney, Agent or Firm:
HANGZHOU TIANQIN INTELLECTUAL PROPERTY AGENCY CO., LTD. (CN)
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