Monoamine oxidase and application thereof in synthesis of chiral azabicyclic compounds

Abstract

The invention provides a monoamine oxidase with high catalysis activity, strong enantioselectivity and good substrate survivability, and an enzyme-chemical synthesis method using the above enzyme to carry out enzyme catalyzed synthesis of (1S,2S,5R)-6,6-dimethyl-2-substituted-3-azabicyclo[3.1.0]hexane or (1S,3aR,6aS)-1-substituted-octahydrocyclopenta[c]pyrrole in order to further synthesize (1S,2S,5R)-6,6-dimethyl-2-3-azabicyclo[3.1.0]hexane-2-nitrile and its hydrolysate methyl (1S,2S,5R)-6,6-dimethyl-2-3-azabicyclo[3.1.0]hexane-2-carboxylate or further synthesize (1S,3aR,6aS)-1-octahydrocyclopenta[c]pyrrole-1nitrile The invention also provides a gene for coding the monoamine oxidase, a recombinant expression vector containing the gene, a recombinant expression transformant containing the gene, and efficient preparation methods of the vector and the transformant. The monoamine oxidase can respectively react with 3-5 and 5-5 azacyclopentamine systems with different sizes to generate corresponding imides; the imides undergo cyanide addition, so a cyanide is added to an enzyme catalysis system, and the above obtained addition product can be directly alcoholyzed in a hydrochloric acid-alcohol solution to obtain hydrochloride of amino acid methyl ester, so one-pot feeding is realized, the reaction technology is simplified, and industrial production is facilitated.

Description

technical field [0001] The invention belongs to the field of bioengineering, and specifically relates to a monoamine oxidase, a recombinant expression vector and a recombinant expression transformant containing the gene encoding the enzyme, the expressed recombinant enzyme and the preparation method of the recombinant enzyme, and the monoamine oxidase as a catalyst in the synthesis of bocecept Application in Wei intermediates. Background technique [0002] Boceprevir and telaprevir are hepatitis C virus (HCV) protease inhibitors, and their chemical structural formulas are as follows: [0003] [0004] The phase III SPRINT-2 study showed that, compared with standard treatment, 24 weeks of standard treatment with boceprevir can improve the SVR rate in untreated patients with HCV genotype 1. The antiviral efficacy of standard treatment with boceprevir for 24 weeks was similar to that of standard treatment with boceprevir for 44 weeks. [0005] Fragment A is one of the key ...

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Problems solved by technology

[0035] The technical problem to be solved by the present invention is, low yield, high raw material cost, incomplete reaction, and poor corresponding selectivity in the reaction of the reported preparation of boceprevir intermediate fragment A and telaprevir intermediate fragment B. High or unfriendly to the environment, a monoamine oxidase with high catalytic activity, strong enantioselectivity, and good substrate tolerance is provided for enzymatic synthesis of (1S,2S,5R)-6,6-dimethyl -2-substituted-3-azabicyclo[3.1.0]hexane or (1S,3aR,6aS)-1-substituted-octahydrocyclopenta[c]pyrrole, further synthesis of (1S,2S,5R)- 6,6-Dimethyl-3-azabicyclo[3.1.0]hexane-2-carbonitrile and its hydrolysis product (1S,2S,5R)-6,6-dimethyl-3-azabicyclo[ 3.1.0] Hexane-2-methyl carboxylate, or further enzymatic-chemical synthesis of (1S,3aR,6aS)-octahydrocyclopenta[c]pyrrole-1-carbonitrile

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