A Method for Enzymatic Resolution of Chiral Substances

Patent metadata

Field	Value
Publication number	US-2022017935-A1
Application number	US-201917311835-A
Country	US
Kind code	A1
Filing date	Oct 25, 2019
Priority date	Dec 10, 2018
Publication date	Jan 20, 2022
Grant date	—

Abstract

The invention belongs to the field of bioengineering and food technology, and discloses a method for enzymatic resolution of chiral substances, including the following steps: (1) preparing an enzyme solution with a lipase concentration of 1-3000 U/mL, and adding a soluble salt, a hydrophilic solvent and a hydrophobic solvent to the enzyme solution to form a three-liquid phase system; the hydrophobic solvent contains esters or amide compounds composed of racemic chiral compounds; (2) subjecting the three-liquid phase system to enzyme-catalyzed reaction under stirring condition; after the reaction is completed, standing or centrifuging the three-liquid phase system to divide it into three layers, which are a upper liquid layer, a middle liquid layer and a lower liquid layer from top to bottom. The optically pure chiral product after hydrolysis is mainly rich in the middle liquid layer or the lower liquid layer, while the upper liquid layer product is another ester or amide product containing an optically pure chiral product. The method has the advantages of low energy consumption, high raw material utilization rate, and mild reaction conditions, and solves the problems of low chiral resolution efficiency, poor chiral selectivity, low recovery rate, and difficulty in industrialization in the existing enzymatic method.

First claim

1 . A method for enzymatic resolution of chiral substances, characterized in that it comprises the following steps:
(1) preparing an enzyme solution with a lipase concentration of 1-3000 U/mL, adding a soluble salt, a hydrophilic solvent and a hydrophobic solvent to the enzyme solution to form a three-liquid phase system; the mass ratios of the soluble salt, the hydrophilic solvent and the hydrophobic solvent to the enzyme solution are 0.1-0.9, 0.1-5 and 0.1-10, respectively; the hydrophobic solvent contains an ester or amide compound composed of a racemic chiral compound; (2) subjecting the three-liquid phase system to enzyme-catalyzed reaction under stirring condition; after the reaction, standing or centrifuging the three-liquid phase system until it is divided into three layers, which are a upper liquid layer, a middle liquid layer and a lower liquid layer from top to bottom; hydrolyzed optically pure chiral product is mainly concentrated in the middle liquid layer or the lower liquid layer, and the upper liquid layer product is another ester or amide product containing an optically pure chiral product.

2 . The method according to claim 1 , characterized in that the hydrophilic solvent in step (1) is one or more of polyethylene glycol, polypropylene glycol, ethanol, n-propanol, isopropanol, n-butanol, isobutanol, ethylene glycol and acetone; or the hydrophilic solvent is one or more of [BMIM]Br, [BMIM]BF 4 , [EMIM]ETSO 4 and [OMIM]Cl.

3 . The method according to claim 1 , characterized in that the soluble salt in step (1) is one or more of sodium citrate, sodium chloride, ammonium sulfate, sodium carbonate, dipotassium hydrogen phosphate, potassium phosphate, potassium dihydrogen phosphate and dipotassium hydrogen phosphate.

4 . The method according to claim 1 , characterized in that the hydrophobic solvent in step (1) is one or more of n-hexane, diethyl ether, isopropyl ether, ethyl acetate, cyclohexanol, petroleum ether, isooctane, benzene and toluene.

5 . The method according to claim 1 , characterized in that the reaction conditions of step (2) are: a temperature of 30-45° C., and a reaction time of 20 min-4 h.

6 . The method according to claim 5 , characterized in that the pH value of the three-liquid phase system in step (1) is 5-9.

7 . The method according to claim 6 , characterized in that the lipase concentration in step (1) is 5-2000 U/mL.

8 . The method according to claim 7 , characterized in that the mass ratios of the soluble salt, hydrophilic solvent and hydrophobic solvent to the enzyme solution are 0.2-0.8, 0.2-0.8 and 0.2-4, respectively.

9 . The method according to claim 1 , characterized in that the ester or amide compound composed of the racemic chiral compound is one or more of racemic methyl mandelate, racemic Naproxen methyl ester, racemic (4-methoxy-phenyl)-1-ethanol acetate, racemic 1-(4-methoxyphenyl) ethanol acetate, and racemic 6-methyl-5-heptenyl-2-ol acetate.

10 . The method according to claim 1 , characterized in that the ester or amide compound composed of the racemic chiral compound accounts for 0.1%-10% of the mass of the hydrophobic solvent, preferably 1%-5%.

11 . The method according to claim 2 , characterized in that the reaction conditions of step (2) are: a temperature of 30-45° C., and a reaction time of 20 min-4 h.

12 . The method according to claim 3 , characterized in that the reaction conditions of step (2) are: a temperature of 30-45° C., and a reaction time of 20 min-4 h.

13 . The method according to claim 4 , characterized in that the reaction conditions of step (2) are: a temperature of 30-45° C., and a reaction time of 20 min-4 h.

14 . The method according to claim 2 , characterized in that the ester or amide compound composed of the racemic chiral compound is one or more of racemic methyl mandelate, racemic Naproxen methyl ester, racemic (4-methoxy-phenyl)-1-ethanol acetate, racemic 1-(4-methoxyphenyl) ethanol acetate, and racemic 6-methyl-5-heptenyl-2-ol acetate.

15 . The method according to claim 3 , characterized in that the ester or amide compound composed of the racemic chiral compound is one or more of racemic methyl mandelate, racemic Naproxen methyl ester, racemic (4-methoxy-phenyl)-1-ethanol acetate, racemic 1-(4-methoxyphenyl) ethanol acetate, and racemic 6-methyl-5-heptenyl-2-ol acetate.

16 . The method according to claim 4 , characterized in that the ester or amide compound composed of the racemic chiral compound is one or more of racemic methyl mandelate, racemic Naproxen methyl ester, racemic (4-methoxy-phenyl)-1-ethanol acetate, racemic 1-(4-methoxyphenyl) ethanol acetate, and racemic 6-methyl-5-heptenyl-2-ol acetate.

17 . The method according to claim 2 , characterized in that the ester or amide compound composed of the racemic chiral compound accounts for 0.1%-10% of the mass of the hydrophobic solvent, preferably 1%-5%.

18 . The method according to claim 3 , characterized in that the ester or amide compound composed of the racemic chiral compound accounts for 0.1%-10% of the mass of the hydrophobic solvent, preferably 1%-5%.

19 . The method according to claim 4 , characterized in that the ester or amide compound composed of the racemic chiral compound accounts for 0.1%-10% of the mass of the hydrophobic solvent, preferably 1%-5%.

20 . The method according to claim 11 , characterized in that the pH value of the three-liquid phase system in step (1) is 5-9.

Assignees

• Univ South China Tech

Inventors

• Li Zhigang
• Su Jinfen
• Yang Bo
• Wang Yonghua
• Li Zhengcheng
• Chen Huayong

Classifications

• C12Y301/01003 Primary

  Triacylglycerol lipase (3.1.1.3) · CPC title

• C12P7/04

  acyclic · CPC title

• C12P7/62 Primary

  Carboxylic acid esters · CPC title

• C12N9/20

  Triglyceride splitting, e.g. by means of lipase · CPC title

• C12P7/42

  Hydroxy-carboxylic acids · CPC title