title: Cloning, expression, purification, and characterization of glutaredoxin from Antarctic sea-ice bacterium Pseudoalteromonas sp. AN178.
abstract: Glutaredoxins (Grxs) are small ubiquitous redox enzymes that catalyze glutathione-dependent reactions to reduce protein disulfide. In this study, a full-length Grx gene (PsGrx) with 270 nucleotides was isolated from Antarctic sea-ice bacterium Pseudoalteromonas sp. AN178. It encoded deduced 89 amino acid residues with the molecular weight 9.8 kDa. Sequence analysis of the amino acid sequence revealed the catalytic motif CPYC. Recombinant PsGrx (rPsGrx) stably expressed in E. coli BL21 was purified to apparent homogeneity by Ni-affinity chromatography. rPsGrx exhibited optimal activity at 30°C and pH 8.0 and showed 25.5% of the activity at 0°C. It retained 65.0% of activity after incubation at 40°C for 20 min and still exhibited 37.0% activity in 1.0 M NaCl. These results indicated that rPsGrx was a typical cold active protein with low thermostability.
authors.affiliation: School of Marine and Technology, Harbin Institute of Technology, Weihai 264209, China.
authors.forename: Quanfu
authors.initials: Q
authors.lastname: Wang
authors.affiliation: School of Marine and Technology, Harbin Institute of Technology, Weihai 264209, China.
authors.forename: Yanhua
authors.initials: Y
authors.lastname: Hou
authors.affiliation: School of Marine and Technology, Harbin Institute of Technology, Weihai 264209, China.
authors.forename: Yonglei
authors.initials: Y
authors.lastname: Shi
authors.affiliation: School of Marine and Technology, Harbin Institute of Technology, Weihai 264209, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Han
authors.affiliation: School of Marine and Technology, Harbin Institute of Technology, Weihai 264209, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Chen
authors.affiliation: School of Marine and Technology, Harbin Institute of Technology, Weihai 264209, China.
authors.forename: Zhiguo
authors.initials: Z
authors.lastname: Hu
authors.affiliation: Shandong Provincial Engineering Technology Research Center of Marine Health Food (Taixiang Group), Rongcheng 264300, China.
authors.forename: Yuanping
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Shandong Provincial Key Laboratory of Processing Technology of Frozen Prepared Food (Taixiang Group), Rongcheng 264300, China.
authors.forename: YuJin
authors.initials: Y
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000864:Antarctic Regions; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D054477:Glutaredoxins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D007053:Ice; D008969:Molecular Sequence Data; D031161:Pseudoalteromonas / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D054477:Glutaredoxins; D007053:Ice; D011994:Recombinant Proteins
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