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

Patent metadata

Field	Value
Publication number	US-10526622-B2
Application number	US-201815871039-A
Country	US
Kind code	B2
Filing date	Jan 14, 2018
Priority date	May 26, 2017
Publication date	Jan 7, 2020
Grant date	Jan 7, 2020

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 method for preparing (R)-3-hydroxy-5-hexanoate, comprising:
subjecting 3-carbonyl-5-hexanoate (II) to asymmetric reduction in the presence of the ketoreductase of SEQ ID NO: 1 as a catalyst to produce (R)-3-hydroxy-5-hexanoate (I), as shown in the following reaction scheme:

wherein R is alkyl or cycloalkyl having 1 to 8 carbon atoms, or mono- or poly-substituted aryl or aralkyl.

2. The 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).

3. The method according to claim 2 , wherein
the reaction system further comprises a regeneration system for regenerating NADPH from the NADP+.

4. The method according to claim 3 , 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.

5. The method according to claim 4 , 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).

6. The method according to 1 , wherein
a reaction temperature is 15° C. to 35° C.

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

8. The method according to claim 1 , wherein
the initial reaction system further comprises isopropanol, and a concentration percentage of the 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

• G01N30/7206

  interfaced to gas chromatograph (interfaces in general for introducing or extracting samples to be analysed with specially adapted mass spectrometer, see H01J49/04) · CPC title

• C12P7/62 Primary

  Carboxylic acid esters · CPC title

• C07C69/732

  of unsaturated hydroxy carboxylic acids · CPC title

• C12Y101/0108

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

• C12N9/0006

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