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Construction of cofactor self- sufficient whole -cell biocatalyst system for production of S-2-Amino-1-phenylethanol*(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

Issue:
2020 01
Page:
1-12
Research Field:
Articles
Publishing date:

Info

Title:
Construction of cofactor self- sufficient whole -cell biocatalyst system for production of S-2-Amino-1-phenylethanol*
Author(s):
ZHAO Youxi12 LIU Song2 ZHANG Xian2 XU Meijuan2 YANG Taowei2 & RAO Zhiming2**
1 Biochemical Engineering College, Beijing Union University, Beijing 100023, China
2 Ministry of Education Key Laboratory of Industrial Biotechnology, School of Biotechnology, Jiangnan University, Wuxi 214122, China
Keywords:
S-2-Amino-1-phenylethanol phenyl-12-ethanediol -transaminase cofactor self-sufficient whole-cell biocatalyst
CLC:
-
PACS:
DOI:
10.19675/j.cnki.1006-687x.2019.04029
DocumentCode:

Abstract:
S-2-Amino-1-phenylethanol is an important pharmaceutical intermediate and is widely used in the synthesis of various physiologically active drugs. At present, its synthesis is mainly through chemical methods , although the process is mature, the reaction conditions are severe and many by-products are formed. Based on the newly identified ω-transaminase derived from Pseudomonas aeruginosa PAK, a catalyzed enzyme catalytic system combining alcohol dehydrogenase derived from Mycobacterium neoaurum was designed, which synthesized S-2-Amino-1-phenylethanol by one-step biocatalysis of phenyl-1,2-ethanediol. For cofactor regeneration and amino donor source, an interesting point was involved as that a cofactor self-sufficient system was designed by expression of GluDH. By introducing GluDH, a closed cofactor self- sufficient system was constructed, which could realize the simult aneous regeneration of coenzyme (NADP+) and cosubstrate ( L-Glu). The whole-cell transformation of recombinant Escherichia coli BL21 (MPG) showed that in 100 mL NH 4Cl reaction system, after 10 h reaction at pH 8.0 and 37 °C, the recombinant strain with OD600 nm = 30 could catalyze 50 mmol/L phenyl-1,2-ethanediol produces optically pure S-2-amino-1-phenylethanol with a conversion of 99.6%, ee > 99%. This work provides an effective method for catalyzing the synthesis of optically pure S-2-Amino-1-phenylethano from cheap substrates without cofactor supply of and no accumulation of by-products.

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Last Update: 2019-06-19