Method for synthesizing (R)-1,3-butanediol with whole cells of microorganism

Abstract

The invention discloses a method for synthesizing (R)-1,3-butanediol with whole cells of a microorganism, and belongs to the technical field of bio-catalytic asymmetric conversion. According to the method, a microorganism, namely Pichia kudriavzevii CCTCC M 2019329, capable of realizing highly selective whole-cell synthesis of (R)-1,3-butanediol is obtained by screening, whole cells of the microorganism are used as a catalyst, 4-hydroxyl-2-butanone is used as a substrate, the reaction pH, temperature, revolution speed, and auxiliary substrate obtained after single-factor optimization are used,the yield of (R)-1,3-butanediol is up to 80% or more, the optical purity is up to 99% or more, and the product concentration is up to 100 g / L or more.

Description

technical field [0001] The invention relates to a method for synthesizing (R)-1,3-butanediol by whole cells of microorganisms, and belongs to the technical field of biocatalytic asymmetric transformation. Background technique [0002] The chemical structure of (R)-1,3-butanediol ((R)-1,3-BDO) is: [0003] [0004] (R)-1,3-BDO is an unnatural active alcohol and an important pharmaceutical intermediate. In industrial production, it is usually used as an intermediate in the synthesis of pheromones and pesticides, especially in the preparation of azetidinone derivatives. Azetidinone derivatives are the starting materials for the synthesis of penem and carbapenem antibiotics, and penem antibiotics are important β-lactam antibiotics, which have strong bactericidal activity, low toxicity, It has the advantages of wide clinical indications and excellent efficacy. As the demand for penem and carbapenem antibiotics expands in the global market, the demand for (R)-1,3-BDO also in...

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Problems solved by technology

There have been some reports on the synthesis of (R)-1,3-BDO by biotransformation methods. For example, Atsuyama et al. used K.lactis IFO 1267 and C.utilis IAM 4277 to achieve the substrate 4- Catalytic conversion of hydroxy-2-butanone (4H2B), the optical purity of the product reaches 93% and 94%, but the optical purity of the product does not reach 99%, and it needs complicated separation and purification before it can be used as a raw material for pharmaceutical intermediates; Zheng and Rao et al. used C.krusei ZJB-09162 and P.jadinii HBY61 to catalyze the conversion of 4H2B, the optical purity of the product reached 99%, but the substrate concentration could only reach 45g / L, and the yield was 83.9% and 85.1%; Yamamoto coli cells expressing a specific secondary alcohol dehydrogenase of (S)-1,3-BDO by enantioselection using the racemic (R,S)-1,3-BDO as substrate Preparation of (R)-1,3-BDO by sexual oxidation, the optical purity is 95%, but the yield is only 43%; glucose is used as the catalytic substrate, recombinant bacteria are constructed by genetic engineering, and the metabolic regulation of recombinant strains is synthesized (R)-1,3-BDO, Kataoka et al. used this method to catalyze the conversion of glucose, and the product concentration can only reach 9.05g / L, which is far from meeting the requirements of industrialization

Currently, the efficiency of biochemical transformations and the scale of catalytic systems are still limited in terms of (R)-1,3-BDO synthesis

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