Characterization of (R)-selective amine transaminases identified by in silico motif sequence blast

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	Characterization of (R)-selective amine transaminases identified by in silico motif sequence blast
	Jiang, Jinju 1,2,3; Chen, Xi 1,2; Zhang, Dalong 1,2; Wu, Qiaqing 1,2; Zhu, Dunming 1,2
刊名	APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
	2015-03-01
卷号	99 期号:6 页码:2613-2621
关键词	omega-Transaminase Chiralamine Unnatural amino acid Biocatalysis
英文摘要	Compared to (S)-selective amine transaminase ((S)-AT), the (R)-selective counterpart ((R)-AT) has been less studied. As such, a simplified "Motif Sequence Blast" search (Hohne et al. Nat Chem Biol 6:807-813, 2010) was carried out to identify new (R)-ATs from the protein databases. The combined conserved sequence motifs of (R)-ATs based on the previous in silico method of predicting (R)-selective amine transaminase were used as the template sequence for BLASTP search at default settings in NCBI, and six candidate sequences were identified. These putative (R)-AT genes were synthesized and overexpressed in Escherichia coli. Among them, five new (R)-ATs were expressed as soluble protein and showed unusual substrate specificity and high stereoselectivity. Furthermore, several unnatural amino acids, such as d-alanine, d-2-aminobutyric acid, and d-norvaline, were synthesized via the (R)-AT-catalyzed amino transfer reaction to the corresponding keto acids. Optically pure (S)-amines were also obtained by kinetic resolution of racemic amines catalyzed with these new (R)-ATs. Therefore, the Motif Sequence Blast search offers a quick and effective method for in silico identification of new (R)-ATs, and the newly identified (R)-ATs are attractive additions to the toolbox of (R)-ATs for further study and industrial application.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biotechnology & Applied Microbiology
研究领域[WOS]	Biotechnology & Applied Microbiology
关键词[WOS]	SUBSTRATE-SPECIFICITY ; PYRUVATE TRANSAMINASE ; ASYMMETRIC-SYNTHESIS ; VIBRIO-FLUVIALIS ; AMINOTRANSFERASE ; CLONING ; PURIFICATION ; KETONES ; ACIDS ; SITE
收录类别	SCI
语种	英语
WOS记录号	WOS:000350362500013
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/834782/1436]
专题	天津工业生物技术研究所_生物催化与绿色化工朱敦明_期刊论文
作者单位	1.Chinese Acad Sci, Natl Engn Lab Ind Enzymes, Tianjin 300308, Peoples R China 2.Chinese Acad Sci, Tianjin Engn Ctr Biocatalyt Technol, Tianjin Inst Ind Biotechnol, Tianjin 300308, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Jiang, Jinju,Chen, Xi,Zhang, Dalong,et al. Characterization of (R)-selective amine transaminases identified by in silico motif sequence blast[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2015,99(6):2613-2621.
APA	Jiang, Jinju,Chen, Xi,Zhang, Dalong,Wu, Qiaqing,&Zhu, Dunming.(2015).Characterization of (R)-selective amine transaminases identified by in silico motif sequence blast.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,99(6),2613-2621.
MLA	Jiang, Jinju,et al."Characterization of (R)-selective amine transaminases identified by in silico motif sequence blast".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 99.6(2015):2613-2621.