title: Expression and Characterization of a Novel Glycerophosphodiester Phosphodiesterase from Pyrococcus furiosus DSM 3638 That Possesses Lysophospholipase D Activity.
abstract: Glycerophosphodiester phosphodiesterases (GDPD) are enzymes which degrade various glycerophosphodiesters to produce glycerol-3-phosphate and the corresponding alcohol moiety. Apart from this, a very interesting finding is that this enzyme could be used in the degradation of toxic organophosphorus esters, which has resulted in much attention on the biochemical and application research of GDPDs. In the present study, a novel GDPD from Pyrococcus furiosus DSM 3638 (pfGDPD) was successfully expressed in Escherichia coli and biochemically characterized. This enzyme hydrolyzed bis(p-nitrophenyl) phosphate, one substrate analogue of organophosphorus diester, with an optimal reaction temperature 55 °C and pH 8.5. The activity of pfGDPD was strongly dependent on existing of bivalent cations. It was strongly stimulated by Mn(2+) ions, next was Co(2+) and Ni(2+) ions. Further investigations were conducted on its substrate selectivity towards different phospholipids. The results indicated that except of glycerophosphorylcholine (GPC), this enzyme also possessed lysophospholipase D activity toward both sn1-lysophosphatidylcholine (1-LPC) and sn2-lysophosphatidylcholine (2-LPC). Higher activity was found for 1-LPC than 2-LPC; however, no hydrolytic activity was found for phosphatidylcholine (PC). Molecular docking based on the 3D-modeled structure of pfGDPD was conducted in order to provide a structural foundation for the substrate selectivity.
authors.affiliation: School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, China. sdzbwfh@126.com.
authors.forename: Fanghua
authors.initials: F
authors.lastname: Wang
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, China. llhhit@163.com.
authors.forename: Linhui
authors.initials: L
authors.lastname: Lai
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, China. yhlscut@163.com.
authors.forename: Yanhua
authors.initials: Y
authors.lastname: Liu
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, China. yangbo@scut.edu.cn.
authors.forename: Bo
authors.initials: B
authors.lastname: Yang
authors.affiliation: School of Food Science and Engineering, South China University of Technology, Guangzhou 510640, China. yonghw@scut.edu.cn.
authors.forename: Yonghua
authors.initials: Y
authors.lastname: Wang
mesh_terms: D019843:Archaeal Proteins / Q000235:genetics / Q000378:metabolism; D020134:Catalytic Domain; D007700:Kinetics; D008958:Models, Molecular; D062105:Molecular Docking Simulation; D010743:Phospholipids / Q000378:metabolism; D010727:Phosphoric Diester Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D020027:Pyrococcus furiosus / Q000201:enzymology* / Q000235:genetics; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity; D000099189:Lysophospholipase D
chemical_list: D019843:Archaeal Proteins; D010743:Phospholipids; D010727:Phosphoric Diester Hydrolases; D000099189:Lysophospholipase D; C038133:glycerophosphodiester phosphodiesterase
keywords: Pyrococcus furiosus DSM 3638; biochemical properties; glycerophosphodiester phosphodiesterase; lysophospholipase D activity; substrate selectivity