Novel cyanohydrin enzyme and method used for synthesizing chiral cyanohydrin

A cyanohydrinase, a new type of technology, applied in the biological field, can solve the problems of being unsuitable for industrial production, prolonging the time of chiral cyanohydrin, and increasing the difficulty of chiral cyanohydrin steps, so as to shorten the time and reduce the difficulty of preparation.

Inactive Publication Date: 2019-06-14
南京普瑞特生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] But it still has some disadvantages when it is actually used, as it needs to be dried and concentrated when processing the chiral cyanohydrin semi-finished product, that is, two steps are carried out, which prolongs the time

Method used

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  • Novel cyanohydrin enzyme and method used for synthesizing chiral cyanohydrin
  • Novel cyanohydrin enzyme and method used for synthesizing chiral cyanohydrin

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Embodiment 1

[0025] A novel cyanohydrinase, wherein the novel cyanohydrinase is mutated at amino acid residue 103, amino acid residue 128, amino acid residue 141 and amino acid residue 220 of wild cyanohydrin lyase produced, and amino acid residue 103 is mutated to alanine, arginine, asparagine, or aspartic acid, and amino acid residue 128 is mutated to alanine, cysteine, or glutamine, Amino acid residue 141 is mutated to serine, glutamic acid or lysine, and amino acid residue 220 is mutated to leucine, serine or tryptophan.

[0026] The specific preparation method of the novel cyanohydrinase comprises the following steps:

[0027] S1: Cultivate the host cells under suitable conditions, so as to express the novel cyanhydrinase;

[0028] S2: Isolating the novel cyanhydrinase.

[0029] The wild cyanohydrin lyase is set to cassava cyanohydrin lyase MeHNL;

[0030] The 103rd amino acid residue is mutated to asparagine, the 128th amino acid residue is mutated to alanine, the 141st amino acid...

Embodiment 2

[0039] A novel cyanohydrinase, wherein the novel cyanohydrinase is mutated at amino acid residue 103, amino acid residue 128, amino acid residue 141 and amino acid residue 220 of wild cyanohydrin lyase produced, and amino acid residue 103 is mutated to alanine, arginine, asparagine, or aspartic acid, and amino acid residue 128 is mutated to alanine, cysteine, or glutamine, Amino acid residue 141 is mutated to serine, glutamic acid or lysine, and amino acid residue 220 is mutated to leucine, serine or tryptophan.

[0040] The specific preparation method of the novel cyanohydrinase comprises the following steps:

[0041] S1: Cultivate the host cells under suitable conditions, so as to express the novel cyanhydrinase;

[0042] S2: Isolating the novel cyanhydrinase.

[0043] The wild cyanohydrin lyase is set to cassava cyanohydrin lyase MeHNL;

[0044] The 103rd amino acid residue is mutated to asparagine, the 128th amino acid residue is mutated to alanine, the 141st amino acid...

Embodiment 3

[0053] A novel cyanohydrinase, wherein the novel cyanohydrinase is mutated at amino acid residue 103, amino acid residue 128, amino acid residue 141 and amino acid residue 220 of wild cyanohydrin lyase produced, and amino acid residue 103 is mutated to alanine, arginine, asparagine, or aspartic acid, and amino acid residue 128 is mutated to alanine, cysteine, or glutamine, Amino acid residue 141 is mutated to serine, glutamic acid or lysine, and amino acid residue 220 is mutated to leucine, serine or tryptophan.

[0054] The specific preparation method of the novel cyanohydrinase comprises the following steps:

[0055] S1: Cultivate the host cells under suitable conditions, so as to express the novel cyanhydrinase;

[0056] S2: Isolating the novel cyanhydrinase.

[0057] The wild cyanohydrin lyase is set to cassava cyanohydrin lyase MeHNL;

[0058] The 103rd amino acid residue is mutated to asparagine, the 128th amino acid residue is mutated to alanine, the 141st amino acid...

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Abstract

The invention discloses a novel cyanohydrin enzyme and a method used for synthesizing chiral cyanohydrin, and particularly relates to the technical field of biology. The novel cyanohydrin enzyme is generated through the mutation of amino acid residues at the 103 site, the 128 site, the 141 site and the 220 site of a wild cyanohydrin lyase, wherein the amino acid residue at the 103 site is mutated into alanine, arginine, asparagine or asparaginic acid, and the amino acid residue at the 128 site is mutated into alanine, cysteine or glutamine. The novel cyanohydrin enzymeand the method used for synthesizing the chiral cyanohydrin have the advantages that a free drying method is adopted for replacing a stepped drying and concentration method in the prior art for processing semi-finished chiral cyanohydrin, in the actual application, the time required for preparing the chiral cyanohydrin is effectively shortened, the preparation difficulty of the semi-finished chiral cyanohydrin is reduced, and the method can be effectively applied to the industrial production while being applied to the laboratory preparation.

Description

technical field [0001] The invention relates to the field of biotechnology, more specifically, the invention relates to a novel cyanohydrinase and a method for synthesizing chiral cyanohydrin. Background technique [0002] Chiral cyanohydrins and their esters are a class of chiral sources with important academic value in modern asymmetric synthesis research. In addition, it can be easily converted into chiral α-hydroxy acids (esters), α-amino acids, β-amino alcohols and other commercially important pharmaceutical and pesticide intermediates. Therefore, its synthesis has been highly valued by academia and industry. The synthesis of chiral cyanohydrins has always been a challenging subject in the field of asymmetric synthesis. The asymmetric synthesis of chiral cyanohydrins generally requires chiral catalysts: chemical chiral catalysts or biocatalysts. The latter has been paid attention to due to its rich sources, high catalytic efficiency, mild reaction conditions, high yi...

Claims

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Application Information

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IPC IPC(8): C12N9/88C12N15/60
Inventor 刘志斌刘经辉吴庆斌周敦秀
Owner 南京普瑞特生物科技有限公司
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