Preparation method for (r)-3-hydroxyl-5-hexenoate

Patent metadata

Field	Value
Publication number	US-2018340196-A1
Application number	US-201815871039-A
Country	US
Kind code	A1
Filing date	Jan 14, 2018
Priority date	May 26, 2017
Publication date	Nov 29, 2018
Grant date	—

Abstract

The present disclosure relates to the technical field of biochemical engineering and particularly discloses a preparation method for (R)-3-hydroxyl-5-hexenoate. In the method of the present disclosure, the (R)-3-hydroxyl-5-hexenoate is prepared by catalytic reduction of 3-carbonyl-5-hexenoate by ketoreductase with 3-carbonyl-5-hexenoate as the substrate. The amino acid sequence of ketoreductase is shown in SEQ ID NO.1. In the present disclosure, the (R)-3-hydroxyl-5-hexenoate having a very high chiral purity is obtained by asymmetric reduction by ketoreductase as the biocatalyst. The present disclosure has the advantages of easy operation, mild reaction conditions, high reaction yield and good practical industrial application value.

First claim

What is claimed is:

1 . A preparation method for (R)-3-hydroxyl-5-hexenoate, characterized in that a chemical formula (I) of the (R)-3-hydroxyl-5-hexenoate is

wherein R is alkyl or cycloalkyl having 1 to 8 carbon atoms, or mono- or poly-substituted aryl or aralkyl, wherein (R)-3-hydroxyl-5-hexenoate (I) is prepared by asymmetric reduction of 3-carbonyl-5-hexenoate (II) by ketoreductase as catalyst.

2 . The preparation method according to claim 1 , wherein
an amino acid sequence of the ketoreductase is shown in SEQ ID NO.1: or is a mutated amino acid sequence which is obtained by insertion, deletion or substitution of at least one amino acid in the amino acid sequence shown in SEQ ID NO. 1 under the premise of keeping the catalytic activity of the ketoreductase.

3 . The preparation method according to claim 1 , wherein
coenzyme NADP+ is further added in a reaction system, and a dosage of the NADP+ is 0.005% to 0.01% of a dosage (w/w) of the 3-carbonyl-5-hexenoate (II).

4 . The preparation method according to claim 3 , wherein
the reaction system further comprises a regeneration system for regenerating NADPH from the NADP+.

5 . The preparation method according to claim 4 , wherein
the regeneration system of the NADPH in the reaction system is isopropanol dehydrogenase having an amino acid sequence shown in SEQ ID NO.2 or a mutated amino acid sequence obtained by insertion, deletion or substitution of at least one amino acid in the amino acid sequence shown in SEQ ID NO.2 under the premise of keeping the catalytic activity of the isopropanol dehydrogenase.

6 . The preparation method according to claim 5 , wherein
in an initial reaction system, a concentration percentage by mass of the 3-carbonyl-5-hexenoate (II) is 1% to 20%; the dosage of the ketoreductase by wet cell weight is 1% to 30% of the mass of the 3-carbonyl-5-hexenoate (II), and the dosage of the isopropanol dehydrogenase by wet cell weight is 1% to 15% of the mass of the 3-carbonyl-5-hexenoate (II).

7 . The preparation method according to 1 , wherein
the reaction temperature in the reaction system is 15° C. to 35° C.

8 . The preparation method according to claim 7 , wherein
the pH value of the reaction system is 6 to 9.

9 . The preparation method according to claim 1 , wherein
in the initial reaction system, a concentration percentage of isopropanol is 5% to 20% (v/v).

Assignees

• Univ Fudan

Inventors

• Chen Fener
• Huang Zedu
• Meng Ge
• Liu Minjie
• Li Zhining
• Wang Zexu
• Peng Haihui
• Xiong Fangjun
• Wu Yan
• Tao Yuan

Classifications

• C12Y101/0108

  Isopropanol dehydrogenase (NADP+) (1.1.1.80) · CPC title

• C12N9/0006

  acting on CH-OH groups as donors (1.1) · CPC title

• G01N30/88

  Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 · CPC title

• C07C67/28

  by modifying the hydroxylic moiety of the ester, such modification not being an introduction of an ester group · CPC title

• C12P7/62 Primary

  Carboxylic acid esters · CPC title