논문 (학술지)
Mutational analyses for product specificity of YjiC towards alpha-mangostin mono-glucoside
| 등록번호 | RPMS-2019-0190788172 | SCI 구분
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※구분 : SCI(SCIE포함), 비SCI |
SCI |
|---|---|---|---|
| 저자명 (주·공동저자) | Kim Tae-Su; Tuoi Thi Le; Hue Thi Nguyen; Cho Kye Woon; Sohng Jae Kyung | ||
| 논문구분 | 국외전문학술지 | 학술지명 | ENZYME AND MICROBIAL TECHNOLOGY |
| ISSN | 0141-0229 | 학술지 출판일자 | - |
| 학술지 볼륨번호 | 118 | 논문페이지 | 76 ~ 82 |
| 학술지 임팩트팩터 | 2.502 | 기여율 | 50 % |
| DOI | 10.1016/j.enzmictec.2018.08.001 | ||
| 초록 | Abstract Glycosyltransferases (GTs) are key enzymes for the post-modification of secondary metabolites in drug development processes. In our prior research, an one-pot enzymatic system produced α-mangostin 3,6-di-O-beta-D-glucopyranoside (Mg1) at a higher proportion using wild-type glycosyltransferase (YjiC) but α-mangostin 3-O-beta-D-glucopyranoside (Mg2) exhibited markedly higher anti-bacterial activities. This study focuses on a Bacillus licheniformis -originated flexible glycosyltransferase by mutagenesis to examine the active site residues involved in glycosylation for a product specificity towards Mg2. The generated H298A, H298S, and H298C mutants of YjiC exhibited a regiospecificity towards glycosylated product (Mg2) and were targeted in this study. The production pattern of Mg1 decreased to 63 (H298A), 85 (H298S) and 95% (H298C) yields compared to the wild-type YjiC. The increase of uridine 5′-diphosphate (UDP) leading to the inhibition of enzyme activity and production of uridine 5′-diphosphate glucose (UDP-glucose) in overall system was critical for the specific glycosylated product formation rate. H298A, H298S, and H298C mutants and YjiC exhibited 244, 251, and 186% increases in Mg2 production yields, respectively. And also H298A, H298S, and H298C showed 281, 279, and 251% increases in yield of Mg3 compared with wild type YjiC, respectively. There was improved conversion of both mono-glucosides product (Mg2a and Mg3) than di-glucosides products. The H298 mutants were found to overcome the limitation of the wild-type YjiC for regioselective synthesis of Mg2 by an enzymatic system. Highlights H298 mutants can produce mono-glucoside because of a reduced deglycosylation activity and UDP inhibition. H298A, H298S, and H298C mutants and YjiC exhibited 244, 251, and 186% increases in Mg2 production yields, respectively. Engineered YjiC can be utilized for regiospecific glycosylation of numerous chemical structure on polyketides, flavonoids etc. | ||
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