Method for the asymmetric synthesis of (2s,3r)-p-thiamphenylphenylserine in whole cells with "multiple enzymes in one bacterium"

Abstract

The invention provides a method for asymmetrically synthesizing (2S, 3R)-p-thiamphenicol phenylserine in whole cells with "one bacterium and multiple enzymes", which belongs to the field of biocatalysis and enzyme engineering. By simultaneously expressing L-threonine transaldolase (PsLTTA), alcohol dehydrogenase (ApADH) and formate dehydrogenase (CbFDH) in E. Asymmetric and efficient biosynthesis of sulfonylphenylserine. The present invention can obtain the key chiral building block (2S, 3R)-p-thiamphenicol-phenylserine of β-aminoalcohol antibiotics through one-step reaction under normal temperature and pressure, and at the same time remove the inhibitory effect of by-product acetaldehyde on PsLTTA , is an efficient green biosynthetic pathway.

Description

technical field [0001] The invention belongs to the field of biocatalysis and enzyme engineering, and specifically relates to a method for asymmetrically synthesizing (2S, 3R)-p-thiamphenicol phenylserine in whole cells with "one bacterium and multiple enzymes". Background technique [0002] (2S,3R)-P-thymphenylphenylserine is a key chiral building block for the synthesis of β-aminoalcohol antibiotics florfenicol and thiamphenicol. In 2015, China’s export volume of β-aminoalcohol antibiotics alone exceeded 2,700 tons, a year-on-year increase of 9.2%, and the export value reached 186 million US dollars, ranking among my country’s top 10 export drugs. Therefore, the establishment of a fast, efficient and green synthetic route to (2S,3R)-p-thymphenylphenylserine has broad application prospects. [0003] At present, the main method of large-scale production of (2S,3R)-p-thiamphenicol phenylserine at home and abroad is to use p-thiamphenicol benzaldehyde and glycine as raw mater...

AI Technical Summary

Problems solved by technology

Although the reaction route is simple and suitable for industrial production, there are the following problems: 1. The reaction conditions are severe, and strong acid and strong base are used simultaneously; 2. The expensive chiral resolution reagent D-tartaric acid is required

The operation is cumbersome and time-consuming, the production cost is high, and the method adopts the method of chiral resolution instead of the asymmetric catalytic synthesis method, and the yield of the target product is low, only 50%

[0008] Therefore, there is a lack of a high-efficiency synthesis technology of (2S,3R)-p-thiamphenicol phenylserine that is easy to operate, short in reaction time, does not require enzyme fermentation production and purification preparation process, and is more conducive to industrial production and application.

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