Method for synthesizing chiral 2-amino-1-butanol

A synthetic method, amino technology, applied in biochemical equipment and methods, enzymes, redox enzymes, etc., can solve the problems of large pollution, low safety factor, low yield, etc.

Active Publication Date: 2019-05-31
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The synthetic method of above-mentioned (S)-2-amino-1-butanol, chemical method needs high temperature, high pressure and metal catalyst to utilize hydrogenation reduction to obtain, and pollution is big, and safety factor is low
The chemical resolution method also needs to use a large amount of acid-base and other chemical reagents. Although the enzymatic resolution method has mild reaction conditions and good stereoselectivity, the conversion rate of the resolution method is only 50%, and the yield is low.

Method used

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  • Method for synthesizing chiral 2-amino-1-butanol
  • Method for synthesizing chiral 2-amino-1-butanol
  • Method for synthesizing chiral 2-amino-1-butanol

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0111] Example 1. Preparation of (S)-2-amino-1-butanol by coupling alcohol dehydrogenase or carbonyl reductase to leucine dehydrogenase or its mutant of Geobacillus stearothermophilus

[0112] 1. Preparation of engineering bacteria of recombinant alcohol dehydrogenase or its mutant, amino acid dehydrogenase or its mutant

[0113] The genes encoding the relevant enzymes are synthesized separately (codon optimization is performed with Escherichia coli as the host if necessary), and the synthesized genes are connected to various expression vectors to construct. The expression vectors are various conventional vectors in the art. The vector of the present invention is specifically pET22b(+), after inserting the coding gene of the relevant enzyme after the whole gene synthesis into the restriction site NdeI and XhoI of pET22b(+), and after the sequence verification is correct, the recombinant vector is obtained . The related gene mutants were obtained by site-directed mutagenesis....

Embodiment 2

[0137] Example 2, Preparation of (S)-2-amino-1-butanol by alcohol dehydrogenase or carbonyl reductase coupled transaminase

[0138] One, the preparation of engineering bacteria of recombinant alcohol dehydrogenase or carbonyl reductase, transaminase

[0139] Carry out with reference to Step 1 of Example 1.

[0140] See Table 4 for details of the enzymes and their mutants involved in this example.

[0141] Enzymes involved in table 4 embodiment 2 and mutants thereof

[0142]

[0143]

[0144] Note: The meanings of W1-W8 are the same as those in Table 1. W10 represents ATA-117 transaminase from Codexis; W11 represents transaminase derived from Aspergillus terreus; W12 represents transaminase derived from Aspergillus fumigatus; W13 represents Neosartorya fischeri derived from W14 means transaminase derived from Gibberella zeae; W15 means transaminase derived from Mycobacterium vanbaalenii. Wn-Mn represents a mutant of Wn (n is a natural number). The numbering and speci...

Embodiment 3

[0157] Example 3, Preparation of (R)-2-amino-1-butanol by coupling transaminase with alcohol dehydrogenase or carbonyl reductase

[0158] 1. Preparation of engineering bacteria for recombinant alcohol dehydrogenase and transaminase

[0159] Carry out with reference to Step 1 of Example 1.

[0160] See Table 6 for details of the enzymes and their mutants involved in this example.

[0161] Enzymes involved in table 6 embodiment 3 and mutants thereof

[0162]

[0163]

[0164] Note: The meanings of W1-W8 are the same as those in Table 1. W16 indicates transaminase derived from Bacillus megaterium; W17 indicates transaminase derived from Pseudomonas aeruginosa (P. aeruginosa). Wn-Mn represents a mutant of Wn (n is a natural number). The numbering and specific nomenclature of protein and gene substitutions are the same as in Table 1.

[0165] 2. Expression of recombinant alcohol dehydrogenase and transaminase and preparation of crude enzyme

[0166] Carry out with refer...

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Abstract

The invention discloses a method for synthesizing chiral 2-amino-1-butanol. The method comprises the following steps: 1,2-butanediol is used as a substrate and is catalyzed by enzyme A and coenzyme thereof to generate 2-ketone-1-butanol; the 2-ketone-1-butanol serves as a substrate, and the 2-amino-1-butanol is generated through catalytic reaction of enzyme B and coenzyme thereof. The enzyme A isselected from alcohol dehydrogenase, carbonyl reductase or mutants of the two enzymes; the enzyme B is selected from amino acid dehydrogenases, transaminases or mutants of the two enzymes. The invention provides a brand-new green biosynthesis route. The cheap 1,2-butanediol is used as a raw material, and the chiral 2-amino-1-butanol is synthesized through multi-enzyme co-expression or cascade or step-by-step catalysis, namely (S)-2-amino-1-butanol and (R)-2-amino-1-butanol.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a synthesis method of chiral 2-amino-1-butanol, in particular to a method for synthesizing chiral 2-amino-1-butanol by catalyzing biological enzymes. Background technique [0002] In recent years, the green chemical process centered on the application of biocatalysts has attracted more and more attention, especially the cascade catalytic reactions that use natural enzymes or newly modified enzymes as multi-enzyme molecular machines are favored by researchers and industries (Fischereder et al., ACS Catal.,2016,6(1):23-30; France et al.,ACS Catal.,2016,6(6):3753-3759; Li et al.,2016,J Agr Food Chem ., 64(46):8927-8934.). The new cascade reaction pathway constructed by multi-enzyme molecular machines can not only reduce the loss of intermediates, but also improve the conversion efficiency and greatly reduce the production cost of the process (Schrittwieser et al., Curr Opin Chem Biol., 2...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/00C12N9/04C12N9/06C12N9/10
CPCY02E50/10
Inventor 孙周通赵强刘保艳曲戈
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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