A method for preparing s-4-chloro-3-hydroxybutyric acid ethyl ester by coupling extraction with enzyme membrane reactor

Abstract

The invention discloses a method for preparing ethyl s-4-chloro-3-hydroxybutyrate by coupling extraction with an enzyme membrane reactor. The method comprises the following steps: (1) pumping a fixed liquid into the enzyme membrane reactor; (2) Dissolve s-carbonyl reductase and glucose dehydrogenase in phosphate buffer of pH 7.0, so that the enzymatic activities of both are 10ku / L-100ku / L, then pump into the enzyme membrane reactor, and Circulate at 10-20℃ for 1h-3h; (3) Contain ethyl 4-chloroacetoacetate with a concentration of 5%-30%, glucose with a concentration of 50-300g / L, and glucose with a concentration of 0%-70% The organic solvent mixed solution is pumped into the enzymatic membrane reactor and circulated; (4) 6-24 hours after the enzymatic catalyzed reaction starts, the same organic solvent as in the reaction is introduced into the other side of the membrane and circulated. In the method for preparing ethyl s-4-chloro-3-hydroxybutyrate by coupling extraction with an enzyme membrane reactor, ethyl s-4-chloro-3-hydroxybutyrate is prepared by coupling extraction with an enzyme membrane reactor, which can continuously The collected product can relieve the product's inhibition of enzymes or toxicity to cells.

Description

technical field [0001] The invention relates to the technical field of enzyme engineering, in particular to a method for preparing s-4-chloro-3-hydroxybutyric acid ethyl ester by coupling extraction with an enzyme membrane reactor. Background technique [0002] Ethyl s-4-chloro-3-hydroxybutyrate is an important pharmaceutical intermediate and chiral alcohol. After cyanation, reduction and other reactions, it can be used for the synthesis of many active drugs, and it is also the global sales ranking of synthesis. The key chiral intermediate of the top ten statins Lipitor is in great market demand. [0003] The preparation of ethyl s-4-chloro-3-hydroxybutyrate mainly adopts biological methods. The commonly used method is to use microorganisms to prepare carbonyl reductase and coenzyme regeneration system, and then use these two enzymes to biocatalyze 4-chloroacetoacetate. Ethyl ester is used to synthesize ethyl s-4-chloro-3-hydroxybutyrate. Because ethyl s-4-chloro-3-hydroxyb...

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Problems solved by technology

[0004] However, due to the presence of enzymes or cells, the organic solvent and water will be emulsified into a homogeneous phase, which will cause difficulties in the subsequent extraction process and reduce the yield. Patent document CN104651292A discloses a method using macroporous adsorption resins The method of removing substrate product inhibition, but this method requires a large amount of eluent, and requires resin regeneration, which will generate a large amount of waste water and solid waste, cumbersome operation, and high cost