Enhancing Production of l-Serine by Increasing the glyA Gene Expression in Methylobacterium sp. MB200 |
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Authors: | Peihong Shen Hongjun Chao Chengjian Jiang Zhangde Long Changhao Wang Bo Wu |
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Institution: | 1. College of Life Science and Technology, The Key Laboratory of Ministry of Education for Microbial and Plant Genetic Engineering, Guangxi University, 100 Daxue Road, Nanning, Guangxi, 530005, China
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Abstract: | Microbial fermentation using methylotrophic bacteria is one of the most promising methods for l-serine production. Here we describe the metabolic engineering of a Methylobacterium strain to increase the production of l-serine. The glyA gene, encoding serine hydroxymethyltransferase (SHMT), was isolated from the genomic DNA of Methylobacterium sp. MB200, using a DNA fragment encoding Methylobacterium extorquens AM1 SHMT as a probe, and inserted into the vector pLAFR3. The resulting construct was transformed into Methylobacterium sp. MB200 using triparental mating. The genetic-engineered strain, designated as Methylobacterium sp. MB202, was shown to produce 11.4?±?0.6 mg/ml serine in resting cell reactions from 30 mg/ml wet cells, 20 mg/ml glycine, and 70 mg/ml methanol in 2 days, representing a 4.4-fold increase from that of the wild strain. The results demonstrated the potential for improving l-serine production by manipulating the glyA in bacteria and should facilitate the production of l-serine using Methylobacterium sp. strains. |
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