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首页> 外文期刊>BMC Biotechnology >A novel serine hydroxymethyltransferase from Arthrobacter nicotianae: characterization and improving catalytic efficiency by rational design
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A novel serine hydroxymethyltransferase from Arthrobacter nicotianae: characterization and improving catalytic efficiency by rational design

机译：烟草节杆菌的新型丝氨酸羟甲基转移酶：通过合理设计表征并提高催化效率

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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 An SHMT 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.
机译：背景丝氨酸羟甲基转移酶（SHMT）是L-丝氨酸酶促生产中的关键酶，表明获得具有高活性的SHMT的重要性。结果在此，通过简并寡核苷酸引发的PCR获得了新的SHMT基因glyA，并编码了与大肠杆菌已知的SHMT具有54.3％相似性的新的SHMT。所获得的蛋白质SHMT在40°C和pH≥7.5时表现出最佳活性，并且在弱碱性条件下（pH≤6.5-8.5）较Hyphomicrobiummethylovorum的SHMT（pH≤6.0-7.5）更稳定。为了提高野生型的催化效率，使用了基于序列比对和生物信息学预测的定点诱变，发现突变型I249L的催化效率比野生型高了2.78倍，其中在249位亮氨酸替代异亮氨酸。结论这项研究提供了有关有趣位点的有用信息，以及DOP-PCR在克隆新glyA基因中的应用。

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