Substrate profile of an 蠅-transaminase from Burkholderia vietnamiensis and its potential for the production of optically pure amines and unnatural amino acids

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• 作者：Jinju Jiang ; Xi Chen ; Jinhui Feng ; Qiaqing Wu ; Dunming Zhu
• 关键词：Biocatalysis ; 蠅-Transaminase ; Chiral amine ; Unnatural amino acid ; 尾-Keto ester
• 刊名：Journal of Molecular Catalysis B: Enzymatic
• 出版年：February, 2014
• 年：2014
• 卷：100
• 期：Complete
• 页码：32-39
• 全文大小：922 K

文摘

A new (S)-enantioselective 蠅-transaminase (蠅-TA) gene from Burkholderia vietnamiensis G4 was functionally expressed in Escherichia coli BL21 (DE3), and the purified recombinant N-terminal His-tagged 蠅-TA (HBV-蠅-TA) had a dimeric structure with optimum pH and temperature of 8.4 and 40 掳C, respectively. The enzyme showed higher activities toward aromatic amines than aliphatic amines and (S)-1-methylbenzylamine ((S)-伪-MBA) was the most active amino donor. For amino acceptor, keto acids, keto esters and aldehydes were more reactive than ketones with pyruvate ethyl ester being most active. Several chiral amines and unnatural amino acids or esters were synthesized using HBV-蠅-TA as the catalyst and isopropylamine or (S)-伪-MBA as amino donor. Notably, HBV-蠅-TA catalyzed the amino transfer to 尾-keto esters to give optically pure 尾-amino acid esters. In addition, glyoxylate was used as amino acceptor for the first time in the kinetic resolution of racemic amines and optically pure amines, such as (R)-1-methylbenzylamine, (R)-1-phenylpropylamine, (R)-2-amino-4-phenylbutane and (R)-1-aminotetraline, were obtained.