title: Expression, purification, and characterization of phosphatidylserine synthase from Escherichia coli K12 in Bacillus subtilis.
abstract: Although phosphatidylserine synthase (PSS) from Escherichia coli is an ideal enzyme for phospholipid production, its application in the food industry has been limited because of the low PSS yield. In this study, the pss gene was cloned from E. coli K(12) and expressed in Bacillus subtilis DB104, and the recombinant PSS was characterized subsequently. PSS was purified to 39.59-fold, and the highest activity was detected as 13.62 U/mg. The enzyme was found to be stable in a pH range of 6.5-9.5, with optimal pH values of 8.0 for hydrolysis and 7.0 for transphosphatidylation, respectively. The optimal temperature for PSS activity was 35 degrees C. The enzyme activity could be detected after 1 h of heating at 65 degrees C. Among the detected detergents and metal ions, Triton X-100, Ca(2+), Mn(2+), and Co(2+) could improve PSS activity. The transformation of phosphatidylcholine to phosphatidylserine under PSS catalyzation was carried out in a biphasic system, which confirmed the actual catalyzing ability of the recombinant protein.
authors.affiliation: Tianjin Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, 300457 Tianjin, People's Republic of China.
authors.forename: Ye-Ni
authors.initials: YN
authors.lastname: Zhang
authors.forename: Fu-Ping
authors.initials: FP
authors.lastname: Lu
authors.forename: Guan-Qun
authors.initials: GQ
authors.lastname: Chen
authors.forename: Yu
authors.initials: Y
authors.lastname: Li
authors.forename: Jian-Ling
authors.initials: JL
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000235:genetics*; D010717:CDPdiacylglycerol-Serine O-Phosphatidyltransferase / Q000235:genetics* / Q000302:isolation & purification; D003001:Cloning, Molecular; D004795:Enzyme Stability; D048168:Escherichia coli K12 / Q000201:enzymology*; D015870:Gene Expression; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D014169:Transformation, Bacterial
chemical_list: D011994:Recombinant Proteins; D010717:CDPdiacylglycerol-Serine O-Phosphatidyltransferase