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A kind of recombinant cell and its application in synthesizing α, β-unsaturated aldehydes

A technology for recombining cells and recombining carriers, which is applied in the field of α,β-unsaturated aldehyde synthesis to achieve the effects of improving catalytic efficiency, high selectivity and shortening reaction time

Active Publication Date: 2022-04-19
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the construction of co-expressed enol dehydrogenase YsADH, NADPH oxidase TkNOX and hemoglobin VsHGB, nor has there been any report on the selective oxidation of α ,Report of α,β-unsaturated aldehydes from β-unsaturated alcohols

Method used

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  • A kind of recombinant cell and its application in synthesizing α, β-unsaturated aldehydes
  • A kind of recombinant cell and its application in synthesizing α, β-unsaturated aldehydes
  • A kind of recombinant cell and its application in synthesizing α, β-unsaturated aldehydes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Acquisition of Enol Dehydrogenase Encoding Gene of Yorkella sp.WZY002

[0039] Using the published enol dehydrogenase (YsADH) encoding gene (GenBank accession number KF887947) from Yorkella sp. WZY002, after codon optimization, artificial synthesis (Suzhou Jinweizhi Biotechnology Co., Ltd. Provide gene synthesis service) The gene encoding enol dehydrogenase, the amino acid sequence and nucleotide sequence are respectively shown in SEQ ID NO.1 and SEQ ID NO.2.

Embodiment 2

[0040] Example 2 Acquisition of gene encoding NADPH oxidase from Thermococcus kodakaraensis

[0041] Using the published gene encoding NADPH oxidase (TkNOX) from Thermococcus kodakaraensis (GenBank accession number is BAD85488), after codon optimization, artificially synthesized (Gene synthesis service provided by Hangzhou Qingke Biotechnology Co., Ltd.) The NADPH oxidase (TkNOX) coding gene, the amino acid sequence and nucleotide sequence are respectively shown in SEQ ID NO.3 and SEQ ID NO.4.

Embodiment 3

[0042] Example 3 Acquisition of the gene encoding hemoglobin from Vitreoscilla stercoraria

[0043]Using the published gene encoding hemoglobin (VsHGB) from Vitreoscilla stercoraria (GenBank accession number is AAA27584), after codon optimization, artificially synthesized (Gene synthesis service provided by Hangzhou Qingke Biotechnology Co., Ltd.) Myoglobinase (VsHGB) encoding gene, the amino acid sequence and nucleotide sequence are respectively shown in SEQ ID NO.5 and SEQ ID NO.6.

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Abstract

The invention discloses a recombinant cell and its application for synthesizing α, β-unsaturated aldehydes. The recombinant cell is constructed by introducing enol dehydrogenase encoding gene, NADPH oxidase encoding gene and hemoglobin encoding gene into host cells. to make. The present invention fuses and co-expresses enol dehydrogenase, NADPH oxidase and hemoglobin recombinant cells to catalyze the selective oxidation of 200mM 2-butene-1-alcohol in a one-pot cascade method, and the substrate is completely converted into the product 2-1-alcohol after 6 hours of reaction Crotonaldehyde significantly improves the catalytic efficiency of genetically engineered bacteria; the method has high selectivity, no overoxidation product saturated aldehyde, enoic acid and saturated acid is detected in the reaction process, and the generation of by-products is avoided; except oxygen , does not need to add other auxiliary substrates, and does not need to add catalase; the method is green and efficient, and is suitable for large-scale industrial production.

Description

(1) Technical field [0001] The invention relates to the synthesis of α, β-unsaturated aldehydes, in particular to a recombinant cell co-expressing enol dehydrogenase, NADPH oxidase and hemoglobin and its application in synthesizing α, β-unsaturated aldehydes. (2) Background technology [0002] α,β-Unsaturated aldehydes are commonly used in the perfume industry and are also important intermediates in organic synthesis. For example, citral (3,7-dimethyl-2,6-octadiene-1-al) is used in the production of ionones, vitamins A and E, and carotenoids, among others. α,β-Unsaturated aldehydes can be obtained by selective oxidation of corresponding α,β-unsaturated alcohols by chemical catalysis or biocatalysis. Chemical catalysis usually suffers from excessive oxidation and poor selectivity. In addition, traditional chemical oxidizers have certain dangers and environmental hazards. Correspondingly, biocatalysis has received more and more attention due to its high activity, high selec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12N15/70C12P7/24C12R1/19
CPCC12N9/0006C07K14/805C12N9/0022C12Y106/03001C12N15/70C12P7/24
Inventor 应向贤乔艳汪钊
Owner ZHEJIANG UNIV OF TECH
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