title: Characterization of a serine hydroxymethyltransferase for L-serine enzymatic production from Pseudomonas plecoglossicida.
abstract: Pseudomonas plecoglossicida, a bacterium strain that exhibits high Serine hydroxymethyltransferase (SHMT) activity, was isolated from the seawater. A full-length glyA encoding SHMT was obtained by a modified thermal asymmetric interlaced-PCR (TRIL-PCR), which consisted of 1,254 bp, encoded a 417 amino acid polypeptide, and shared the highest identity (75 %) with a glyA gene from Acinetobacter radioresistens CMC-1. Recombinant glyA gene was expressed in Escherichia coli BL21 (DE3) and purified by electrophoretic homogeneity. The enzyme showed the optimal activity at pH 8.0 and 40 °C, and remained stable in high alkali conditions. Using SHMT to produce L-serine by catalyzing the reaction of glycine and tetrahydrofolate is one of the most promising routes to synthesize L-serine, achieving 33.4 mM L-serine at the 12th h of the enzymatic reaction with the substrates of glycine (133 mM) and formaldehyde (13.3 mM). The properties make the SHMT a candidate for further enzymatic studies and industrial applications.
authors.affiliation: State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China.
authors.forename: Wei
authors.initials: W
authors.lastname: Jiang
authors.forename: Bingzhao
authors.initials: B
authors.lastname: Xia
authors.forename: Junjie
authors.initials: J
authors.lastname: Huang
authors.forename: Ziduo
authors.initials: Z
authors.lastname: Liu
mesh_terms: D000150:Acinetobacter / Q000201:enzymology / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D019143:Evolution, Molecular; D015870:Gene Expression; D005798:Genes, Bacterial; D012696:Glycine Hydroxymethyltransferase / Q000235:genetics* / Q000378:metabolism*; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011549:Pseudomonas / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000378:metabolism; D012623:Seawater / Q000382:microbiology*; D017385:Sequence Homology; D012694:Serine / Q000096:biosynthesis*
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D012694:Serine; D012696:Glycine Hydroxymethyltransferase
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