Title:
CYCLODEXTRIN GLUCOSYLTRANSFERASE WITH IMPROVED SOLVENT TOLERANCE AND PREPARATION THEREOF
Document Type and Number:
WIPO Patent Application WO/2024/045796
Kind Code:
A1
Abstract:
Disclosed are a cyclodextrin glucosyltransferase with improved solvent tolerance and preparation thereof, belonging to the technical field of enzyme engineering and genetic engineering. The present invention constructs four cyclodextrin glucosyltransferase mutants with improved organic solvent tolerance. The mutant with the best tolerance to DMSO and methanol is G539I/R146F/D147N, which has increased tolerance by 1.6 and 1.7 times compared with WT, respectively; the mutant with the best tolerance to ethanol is R146F, which has increased tolerance by 1.4 times compared with WT; the mutant with the best tolerance to acetone is G539I/R146F, which has increased tolerance by 1.5 times compared with the WT. The present invention helps to expand the application of glycosyltransferase in an organic reaction system, improves the enzymatic efficiency of CGTase on a natural hydrophobic substrate, and has great application prospects.
Inventors:
HAN RUIZHI (CN)
NI YE (CN)
JIANG YULIN (CN)
FAN ZHAOYUE (CN)
NI YE (CN)
JIANG YULIN (CN)
FAN ZHAOYUE (CN)
Application Number:
PCT/CN2023/102014
Publication Date:
March 07, 2024
Filing Date:
June 25, 2023
Export Citation:
Assignee:
UNIV JIANGNAN (CN)
International Classes:
C12N9/10; C12N1/21; C12N15/54; C12N15/70; C12P19/18; C12P19/60
Domestic Patent References:
| WO2002002725A1 | 2002-01-10 |
Foreign References:
| CN116144622A | 2023-05-23 | |||
| CN113817704A | 2021-12-21 | |||
| CN103484439A | 2014-01-01 |
Other References:
CHAI BAOCHENG, JIANG YULIN, NI YE, HAN RUIZHI: "Engineering the 182 site of cyclodextrin glucosyltransferase for glycosylated genistein synthesis", CHINESE JOURNAL OF BIOTECHNOLOGY, vol. 38, no. 2, 25 February 2022 (2022-02-25), pages 749 - 759, XP093143704, ISSN: 1872-2075, DOI: 10.13345/j.cjb.210144
KNEGTEL RONALD M A, WIND RICHE `LE D, ¨TTE HENRIE, ROZEBOOM J, KALK KOR H, BUITELAAR REINETTA M, DIJKHUIZEN LUBBERT, DIJKSTRA BAUK: "Crystal Structure at 2.3 Å Resolution and Revised Nucleotide Sequence of the Thermostable Cyclodextrin Glycosyltransferase from Thermoanaerobacterium thermosulfurigenes EM1", JOURNAL OF MOLECULAR BIOLOGY, ACADEMIC PRESS, UNITED KINGDOM, vol. 256, no. 3, 1 March 1996 (1996-03-01), United Kingdom , pages 611 - 622, XP093143707, ISSN: 0022-2836, DOI: 10.1006/jmbi.1996.0113
KNEGTEL RONALD M A, WIND RICHE `LE D, ¨TTE HENRIE, ROZEBOOM J, KALK KOR H, BUITELAAR REINETTA M, DIJKHUIZEN LUBBERT, DIJKSTRA BAUK: "Crystal Structure at 2.3 Å Resolution and Revised Nucleotide Sequence of the Thermostable Cyclodextrin Glycosyltransferase from Thermoanaerobacterium thermosulfurigenes EM1", JOURNAL OF MOLECULAR BIOLOGY, ACADEMIC PRESS, UNITED KINGDOM, vol. 256, no. 3, 1 March 1996 (1996-03-01), United Kingdom , pages 611 - 622, XP093143707, ISSN: 0022-2836, DOI: 10.1006/jmbi.1996.0113
Attorney, Agent or Firm:
HARBIN SHINEIP INTELLECTUAL PROPERTY AGENCY CO., LTD. (CN)
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