Mining of aminotransferase gene ota3 from Bacillus pumilus W3 via genome analysis, gene cloning and expressing for compound bioamination
| Autor: | Zhengbing Guan, Di Meng, Xiangru Liao, Lixin Zhai, Yujie Cai, Lai Yingjie, Tian Qiaopeng, Yang Shaolan |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Hot Temperature Gene Expression Biology Molecular cloning Genome Catalysis Transaminase 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Bacterial Proteins Genetics Cloning Molecular Peptide sequence Pyridoxal Gene Transaminases Bacillus pumilus Molecular mass General Medicine biology.organism_classification Recombinant Proteins 030104 developmental biology chemistry Biochemistry 030220 oncology & carcinogenesis Genome Bacterial |
| Zdroj: | Gene. 686:21-28 |
| ISSN: | 0378-1119 |
| Popis: | Aminotransferases are widely employed as biocatalysts to produce chiral amines and biologically active pharmaceuticals via asymmetric synthesis. In this study, transaminase genes in the Bacillus pumilus W3 genome were analysed, and gene ota3 encoding a putative (R)-selective transaminase was identified. The sequence of ota3 shares highest sequence identity (24.7%) with the first (R)-selective aminotransferase from Arthrobacter sp. KNK 168. Amino acid sequence and conserved domains analyses indicated that ω-BPAT encoded by ota3 belonged to the pyridoxal 5′-phosphate-dependent class IV (PLPDE_IV) superfamily. Both native and codon-optimised ω-BPAT genes were recombinantly expressed, and the purified proteins had a molecular mass of ~33.4 kDa. Furthermore, enantioselectivity tests with (S)- and (R)-α-phenethylamine revealed its (R)-selectivity. The optimal conditions for catalytic reaction were 45 °C and pH 7.0, and ω-BPAT retained stability at 20 °C and pH 7.0. Thus, ω-BPAT is a novel (R)-selective aminotransferase with great potential as a universal biocatalyst. |
| Databáze: | OpenAIRE |
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