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Multienzyme recombinant cell and method of synthesizing D-pantolactone through multienzyme cascade catalysis

A technology of pantolactone and recombinant cells, which is applied in the field of multi-enzyme recombinant cells and its application in the synthesis of D-pantolactone, can solve the problems of industrial application, cumbersome reaction steps and long reaction time to avoid hydrolysis, simplify the reaction process, and reduce production costs

Active Publication Date: 2019-11-08
HANGZHOU XINFU TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the method involves the hydrolysis of the intermediate product ketopantoate lactone, the use of D-ketopantoate reductase and the subsequent acidic cyclization of D-pantoate into D-pantoate lactone, the reaction steps are more cumbersome , consumes more acid, and the reaction time is longer, which is still not up to the requirements of industrial applications
[0004] At present, there is no report on the direct synthesis of D-pantolactone by biological or enzymatic catalyzed deracemization of DL-pantolactone

Method used

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  • Multienzyme recombinant cell and method of synthesizing D-pantolactone through multienzyme cascade catalysis
  • Multienzyme recombinant cell and method of synthesizing D-pantolactone through multienzyme cascade catalysis
  • Multienzyme recombinant cell and method of synthesizing D-pantolactone through multienzyme cascade catalysis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1 Acquisition of gene encoding L-pantolactone dehydrogenase from Amycolatopsis methylophilus

[0068] Using the published oxidase-encoding gene from Amycolatopsis methanolica (GenBank accession number is AIJ22255.1), after codon optimization, it was artificially synthesized (provided by Suzhou Jinweizhi Biotechnology Co., Ltd. Gene synthesis service) L-pantolactone dehydrogenase coding gene, the nucleotide sequence is shown in SEQ ID NO:1.

Embodiment 2

[0069] Example 2 Acquisition of gene encoding D-ketopantolactone reductase from Saccharomyces cerevisiae

[0070] Using the published reductase encoding gene (GenBank accession number: CAA98692.1) derived from Saccharomyces cerevisiae, after codon optimization, artificially synthesized (gene synthesis service provided by Suzhou Jinweizhi Biotechnology Co., Ltd.) D- The gene encoding ketopantolactone reductase has a nucleotide sequence as shown in SEQ ID NO:3.

Embodiment 3

[0071] Example 3 Acquisition of the gene encoding the glucose dehydrogenase of Exiguobacterium

[0072] Using the published dehydrogenase encoding gene (GenBank accession number ACB59697.1) derived from Exiguobacterium sibiricum, after codon optimization, artificially synthesized (Suzhou Jinweizhi Biotechnology Co., Ltd. provides gene synthesis services) micro Bacillus glucose dehydrogenase coding gene, the nucleotide sequence is shown in SEQ ID NO:5.

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PUM

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Abstract

The invention relates to the technical field of microbial genetic engineering, in particular to a recombinant cell and application thereof to catalytic synthesis of D-pantolactone. The recombinant cell conducts induction to generate L-pantolactone dehydrogenase, D-ketopantolactone reductase and glucose dehydrogenase, and DL-pantolactone or L-pantolactone can be directly synthesized into the D-pantolactone. The invention further provides a method for synthesizing the D-pantolactone through multienzyme cascade direct catalysis. Compared with an enzymatic resolution technology applied to the industry, the reaction process is simplified, and the reaction efficiency is improved.

Description

technical field [0001] The invention relates to the technical field of microbial genetic engineering, in particular to a multi-enzyme recombinant cell and its application in synthesizing D-pantolactone. Background technique [0002] D-pantothenolactone is an important intermediate in the synthesis of D-pantothenic acid and D-panthenol. The industrial synthesis of D-pantolactone is a technical route combining chemical method and hydrolytic enzyme resolution method, using cheap isobutyraldehyde and formaldehyde as starting materials, and isobutyraldehyde and formaldehyde are hydroxylated under alkaline and high temperature conditions. Aldehydes are condensed to form hydroxypivalaldehyde; then hydrocyanic acid is added for cyanation, and alcohol cyanation reaction is carried out under acidic conditions to form cyanohydrins; cyanohydrins are hydrolyzed under acidic conditions to obtain DL-pantolactone; D- Pantolactone hydrolase selectively hydrolyzes D-pantolactone to generate ...

Claims

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Application Information

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IPC IPC(8): C12N1/21C12N15/70C12P17/04C12R1/19
CPCC12N9/0006C12N15/70C12P17/04C12Y101/01168C12Y101/99027
Inventor 应向贤汪钊赵嫚程先锋林行殷杭华白彦兵陈梁毛王伟张连春余建新金文究姜伟林娄波汪军刘学愚
Owner HANGZHOU XINFU TECH CO LTD
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