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

BMC Biotechnology

Wei Jiang¹  Lin Chen¹  Nan Hu²  Shaohui Yuan¹  Bin Li¹  Ziduo Liu¹ 

[1] State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, P. R. China

[2] College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211800, P. R. China

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

Others  :  1084468
DOI  :  10.1186/s12896-014-0093-9

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

【 摘 要 】

Background

Serine hydroxymethyltransferase (SHMT) is the key enzyme in L-serine enzymatic production, suggesting the importance of obtaining a SHMT with high activity.

Results

Here, 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.

Conclusions

This research provides useful information about the interesting site, and the application of DOP-PCR in cloning a novel glyA gene.

【 授权许可】

   
2014 Jiang et al.; licensee BioMed Central Ltd.

【 图 表 】

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