DIOXYGENASE MUTANTS AND USE THEREOF IN SYNTHESIZING (2S,3R)-3-HYDROXYPIPECOLIC ACID

Abstract: the present disclosure belongs to the technical field of enzyme engineering and genetic engineering, and relates to dioxygenase mutants and use thereof in synthesizing (2s,3r)-3-hydroxypipecolic acid. by mutating arginine at position 97 and asparagine at position 43 of the l-proline-3-hydroxylase, the present disclosure makes the specific activity of r97m/n43t up to 2.42 times that of wild-type (wt) enzyme; after conditions are optimized, r97m/n43t achieves a molar yield of 8.82 mm at a substrate concentration of 10 mm under crude enzyme conditions, with a molar yield rate of 88%, which is 4.11 times that of the wild-type enzyme, and a yield of (2s,3r)-3-hydroxypipecolic acid produced by r97m/n43t catalyzing the l-pipecolic acid is 1278 mg�l. the mutant with improved catalytic efficiency toward the l-pipecolic acid in the present disclosure is more suitable for research on the production of (2s,3r)-3-hydroxypipecolic acid.

Inventor(s): Yao NIE, Yan XU, Songyin ZHAO, Jiaojiao GUAN

CPC Classification: C12N9/0071 (Enzymes; Proenzymes; Compositions thereof (preparations containing enzymes for cleaning teeth , ; medicinal preparations containing enzymes or proenzymes ; enzyme containing detergent compositions ; {enzymes with nucleic acid structure, e.g. ribozymes, }); Processes for preparing, activating, inhibiting, separating or purifying enzymes (preparation of malt ))

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