A novel serine hydroxymethyltransferase from Arthrobacter nicotianae: characterization and improving catalytic efficiency by rational design

BMC Biotechnology

Research Article

Nan Hu¹  Lin Chen²  Ziduo Liu²  Wei Jiang²  Bin Li²  Shaohui Yuan² 

[1] College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, 211800, Nanjing, P. R. China;State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, 430070, Wuhan, P. R. China;

关键词: Arthrobacter nicotianae;    SHMT;    Characterization;    Site-directed mutagenesis;    Catalytic efficiency;   

DOI  :  10.1186/s12896-014-0093-9

 received in 2014-07-13, accepted in 2014-10-22,  发布年份 2014

来源: Springer

【 摘 要 】

BackgroundSerine hydroxymethyltransferase (SHMT) is the key enzyme in L-serine enzymatic production, suggesting the importance of obtaining a SHMT with high activity.ResultsHere, a novel SHMT gene, glyA, was obtained through degenerate oligonucleotide-primed PCR and encoded a novel SHMT with 54.3% similarity to the known SHMT from Escherichia coli. The obtained protein AnSHMT showed the optimal activity at 40°C and pH 7.5, and was more stable in weakly alkali conditions (pH 6.5-8.5) than Hyphomicrobium methylovorum’s SHMT (pH 6.0-7.5), In order to improve the catalytic efficiency of the wild type, the site-directed mutagenesis based on sequences alignment and bioinformatics prediction, was used and the catalytic efficiency of the mutant I249L was found to be 2.78-fold higher than that of the wild-type, with the replacement of isoleucine by leucine at the 249 position.ConclusionsThis research provides useful information about the interesting site, and the application of DOP-PCR in cloning a novel glyA gene.

【 授权许可】

Unknown   
© Jiang et al.; licensee BioMed Central Ltd. 2014. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.