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A novel serine hydroxymethyltransferase from Arthrobacter nicotianae: characterization and improving catalytic efficiency by rational design.
BMC Biotechnol. 2014 Nov 14; 14:93.BB

Abstract

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.

Authors+Show Affiliations

State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, P. R. China. tianya416@126.com.State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, P. R. China. chenlin19890717@163.com.College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing, 211800, P. R. China. hunan@njtech.edu.cn.State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, P. R. China. 984023989@qq.com.State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, P. R. China. 985927577@qq.com.State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, P. R. China. lzd@mail.hzau.edu.cn.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25394480

Citation

Jiang, Wei, et al. "A Novel Serine Hydroxymethyltransferase From Arthrobacter Nicotianae: Characterization and Improving Catalytic Efficiency By Rational Design." BMC Biotechnology, vol. 14, 2014, p. 93.
Jiang W, Chen L, Hu N, et al. A novel serine hydroxymethyltransferase from Arthrobacter nicotianae: characterization and improving catalytic efficiency by rational design. BMC Biotechnol. 2014;14:93.
Jiang, W., Chen, L., Hu, N., Yuan, S., Li, B., & Liu, Z. (2014). A novel serine hydroxymethyltransferase from Arthrobacter nicotianae: characterization and improving catalytic efficiency by rational design. BMC Biotechnology, 14, 93. https://doi.org/10.1186/s12896-014-0093-9
Jiang W, et al. A Novel Serine Hydroxymethyltransferase From Arthrobacter Nicotianae: Characterization and Improving Catalytic Efficiency By Rational Design. BMC Biotechnol. 2014 Nov 14;14:93. PubMed PMID: 25394480.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A novel serine hydroxymethyltransferase from Arthrobacter nicotianae: characterization and improving catalytic efficiency by rational design. AU - Jiang,Wei, AU - Chen,Lin, AU - Hu,Nan, AU - Yuan,Shaohui, AU - Li,Bin, AU - Liu,Ziduo, Y1 - 2014/11/14/ PY - 2014/07/13/received PY - 2014/10/22/accepted PY - 2014/11/15/entrez PY - 2014/11/15/pubmed PY - 2015/10/6/medline SP - 93 EP - 93 JF - BMC biotechnology JO - BMC Biotechnol. VL - 14 N2 - 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. SN - 1472-6750 UR - https://www.unboundmedicine.com/medline/citation/25394480/A_novel_serine_hydroxymethyltransferase_from_Arthrobacter_nicotianae:_characterization_and_improving_catalytic_efficiency_by_rational_design_ L2 - https://bmcbiotechnol.biomedcentral.com/articles/10.1186/s12896-014-0093-9 DB - PRIME DP - Unbound Medicine ER -