Title:
METHOD FOR EFFICIENT BIOSYNTHESIS OF REBAUDIOSIDE D USING GLYCOSYLTRANSFERASE
Document Type and Number:
WIPO Patent Application WO/2023/087518
Kind Code:
A1
Abstract:
A method for efficient biosynthesis of rebaudioside D (Reb D) using a glycosyltransferase, relating to the field of biocatalytic synthesis. A glycosyltransferase having the activity of catalyzing rebaudioside A (Reb A) to synthesize Reb D is obtained by means of excavation, and a high-catalytic-activity mutant YojK-I241T/G327N is obtained by means of directed evolution. Moreover, a coupling reaction is constructed between the glycosyltransferase mutant YojK-I241T/G327N and a sucrose synthase AtSUSy derived from Arabidopsis thaliana, efficient catalytic synthesis of Reb D is implemented by using Reb A as a substrate, 19.32 g/L (20 mmol/L) Reb A is used as the substrate to react for 15 h, 20.59 g/L Reb D is efficiently synthesized, the yield of Reb D reaches 91.29%, and an efficient and green new way is provided for production of Reb D.
Inventors:
RAO YIJIAN (CN)
GUO BAODANG (CN)
ZHANG YAN (CN)
YUAN ZHENBO (CN)
GUO BAODANG (CN)
ZHANG YAN (CN)
YUAN ZHENBO (CN)
Application Number:
PCT/CN2022/070743
Publication Date:
May 25, 2023
Filing Date:
January 07, 2022
Export Citation:
Assignee:
UNIV JIANGNAN (CN)
International Classes:
C12N9/10; C12N1/21; C12N15/54; C12N15/70; C12P19/18; C12P19/56; C12R1/19
Foreign References:
| CN106350565A | 2017-01-25 | |||
| CN111518782A | 2020-08-11 | |||
| CN110846363A | 2020-02-28 | |||
| CN113462670A | 2021-10-01 | |||
| EP3034614A1 | 2016-06-22 |
Other References:
FEI LIWEN, WANG YONG: "Construction of in situ UDPG Regeneration Strain for Biosynthesis of Rebaudioside D", FOOD SCIENCE, BEIJING FOOD SCIENCE RESEARCH INSTITUTE, CN, vol. 39, no. 22, 1 January 2018 (2018-01-01), CN , pages 116 - 122, XP093068462, ISSN: 1002-6630, DOI: 10.7506/spkx1002-6630-201822018
YANG, YUFENG; FEI, LIWEN; LI, JIANHUA; YANG, JINGYA; WANG, YONG: "Production of Rebaudioside D by Recombinant Escherichia Coli Whole Cell Catalyst", GONGYE WEISHENGWU = INDUSTRIAL MICROBIOLOGY, SHANGHAI GONGYE WEISHENGWU YANJIUSUO, CN, vol. 47, no. 5, 31 October 2017 (2017-10-31), CN , pages 1 - 7, XP009546595, ISSN: 1001-6678, DOI: 10.3969/j.issn.1001-6678.2017.05.001
WANG YU; SUN XUE; JIA XIAOCHEN; ZHU LIPING; YIN HENG: "Comparative transcriptomic of Stevia rebaudiana provides insight into rebaudioside D and rebaudioside M biosynthesis", PLANT PHYSIOLOGY AND BIOCHEMISTRY, ELSEVIER, AMSTERDAM, NL, vol. 167, 17 August 2021 (2021-08-17), AMSTERDAM, NL , pages 541 - 549, XP086819508, ISSN: 0981-9428, DOI: 10.1016/j.plaphy.2021.08.028
YANG, YUFENG; FEI, LIWEN; LI, JIANHUA; YANG, JINGYA; WANG, YONG: "Production of Rebaudioside D by Recombinant Escherichia Coli Whole Cell Catalyst", GONGYE WEISHENGWU = INDUSTRIAL MICROBIOLOGY, SHANGHAI GONGYE WEISHENGWU YANJIUSUO, CN, vol. 47, no. 5, 31 October 2017 (2017-10-31), CN , pages 1 - 7, XP009546595, ISSN: 1001-6678, DOI: 10.3969/j.issn.1001-6678.2017.05.001
WANG YU; SUN XUE; JIA XIAOCHEN; ZHU LIPING; YIN HENG: "Comparative transcriptomic of Stevia rebaudiana provides insight into rebaudioside D and rebaudioside M biosynthesis", PLANT PHYSIOLOGY AND BIOCHEMISTRY, ELSEVIER, AMSTERDAM, NL, vol. 167, 17 August 2021 (2021-08-17), AMSTERDAM, NL , pages 541 - 549, XP086819508, ISSN: 0981-9428, DOI: 10.1016/j.plaphy.2021.08.028
Attorney, Agent or Firm:
HARBIN SHINEIP INTELLECTUAL PROPERTY AGENCY CO., LTD. (CN)
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