Characterization of (R)-selective amine transaminases identified by in silico motif sequence blast | |
Jiang, Jinju1,2,3; Chen, Xi1,2; Zhang, Dalong1,2; Wu, Qiaqing1,2; Zhu, Dunming1,2 | |
刊名 | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
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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. |
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