title: High level expression and characterization of a thermostable lysophospholipase from Thermococcus kodakarensis KOD1.
abstract: Phospholipases can catalyze the hydrolysis of one or more ester and phosphodiester bonds and have a considerable interest in the food, oil leather and pharmaceutical industries. In this report, a lysophospholipase gene from the hyperthermophilic archaeon Thermococcus kodakarensis KOD1 (LysoPL-tk) was cloned. The gene of 783 bp encodes a 260-amino acid protein with a molecular mass of 29 kDa. LysoPL-tk has a consensus motif (GxSxG) and a catalytic triad (S, D, H) of esterases in the deduced amino acid sequence. LysoPL-tk was expressed in Escherichia coli and purified to homogeneity. The enzyme can degrade substrates with both short and long acyl chain lengths. The apparent K                         (m) value for p-nitrophenyl butyrate was 607.1 μM with V                         (max) values of 95.5 U/mg. The enzyme was active at a broad range of pH (5-8) and temperatures (70-95 °C) with the optimum pH and temperature being 8.0 and 85 °C, respectively. The high yield, broad substrate range along with its thermo-stability indicates that LysoPL-tk is a potential enzyme in industrial application.
authors.affiliation: College of Plant Sciences, Jilin University, Changchun 130-062, China.
authors.forename: Zhicheng
authors.initials: Z
authors.lastname: Cui
authors.forename: Yuhan
authors.initials: Y
authors.lastname: Wang
authors.forename: Bang Phuong
authors.initials: BP
authors.lastname: Pham
authors.forename: Fangfang
authors.initials: F
authors.lastname: Ping
authors.forename: Hongyu
authors.initials: H
authors.lastname: Pan
authors.forename: Gang-Won
authors.initials: GW
authors.lastname: Cheong
authors.forename: Shihong
authors.initials: S
authors.lastname: Zhang
authors.forename: Baolei
authors.initials: B
authors.lastname: Jia
mesh_terms: D020816:Amino Acid Motifs; D019843:Archaeal Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008245:Lysophospholipase / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D013379:Substrate Specificity / Q000502:physiology; D019712:Thermococcus / Q000201:enzymology* / Q000235:genetics
chemical_list: D019843:Archaeal Proteins; D011994:Recombinant Proteins; D008245:Lysophospholipase