title: Characterization of an arylsulfatase from a mutant library of Pseudoalteromonas carrageenovora arylsulfatase.
abstract: A library of Pseudoalteromonas carrageenovora arylsulfatase mutants was constructed by introducing random mutagenesis using error-prone PCR. After screening, one mutant strain was obtained whose arylsulfatase had improved thermal stability. Protein sequence analysis revealed one amino acid substitution of H260L. The mutant arylsulfatase (named H260L) retained higher residual activity than wild-type enzyme (named WT) after incubation at 45, 50, 55 and 60°C for 60min. Thermal inactivation analysis showed that the half-life (t1/2) value at 55°C for H260L was 40.6min, while that of WT was 9.1min. When p-nitrophenyl sulfate was used as a substrate, the optimal reaction temperature and pH for the mutant enzyme were 55°C and pH 8.0, respectively. H260L was stable over the pH range of 6.0-9.0. Inhibition assay with EDTA indicated that metal ions play an important role during the catalytic process of the mutant enzyme. The desulfation ratio against agar of Gracilaria lemaneiformis was 82%.
authors.affiliation: College of Food and Biological Engineering, Jimei University, Xiamen 361021, China; Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen 361021, China; Research Center of Food Biotechnology of Xiamen City, Xiamen 361021, China; Key Laboratory of Systemic Utilization and In-depth Processing of Economic Seaweed, Xiamen Southern Ocean Technology Center of China, Xiamen 361021, China.
authors.forename: Yanbing
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: College of Food and Biological Engineering, Jimei University, Xiamen 361021, China.
authors.forename: Han
authors.initials: H
authors.lastname: Liu
authors.affiliation: College of Food and Biological Engineering, Jimei University, Xiamen 361021, China.
authors.forename: Chaochao
authors.initials: C
authors.lastname: Qiao
authors.affiliation: College of Food and Biological Engineering, Jimei University, Xiamen 361021, China; Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen 361021, China; Research Center of Food Biotechnology of Xiamen City, Xiamen 361021, China; Key Laboratory of Systemic Utilization and In-depth Processing of Economic Seaweed, Xiamen Southern Ocean Technology Center of China, Xiamen 361021, China.
authors.forename: Lijun
authors.initials: L
authors.lastname: Li
authors.affiliation: College of Food and Biological Engineering, Jimei University, Xiamen 361021, China; Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen 361021, China; Research Center of Food Biotechnology of Xiamen City, Xiamen 361021, China; Key Laboratory of Systemic Utilization and In-depth Processing of Economic Seaweed, Xiamen Southern Ocean Technology Center of China, Xiamen 361021, China.
authors.forename: Zedong
authors.initials: Z
authors.lastname: Jiang
authors.affiliation: College of Food and Biological Engineering, Jimei University, Xiamen 361021, China; Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen 361021, China; Research Center of Food Biotechnology of Xiamen City, Xiamen 361021, China; Key Laboratory of Systemic Utilization and In-depth Processing of Economic Seaweed, Xiamen Southern Ocean Technology Center of China, Xiamen 361021, China.
authors.forename: Anfeng
authors.initials: A
authors.lastname: Xiao
authors.affiliation: College of Food and Biological Engineering, Jimei University, Xiamen 361021, China; Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen 361021, China; Research Center of Food Biotechnology of Xiamen City, Xiamen 361021, China; Key Laboratory of Systemic Utilization and In-depth Processing of Economic Seaweed, Xiamen Southern Ocean Technology Center of China, Xiamen 361021, China. Electronic address: nihui@jmu.edu.cn.
authors.forename: Hui
authors.initials: H
authors.lastname: Ni
mesh_terms: D000362:Agar / Q000737:chemistry / Q000378:metabolism; D001192:Arylsulfatases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D015723:Gene Library; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D016296:Mutagenesis; D009154:Mutation; D011487:Protein Conformation; D031161:Pseudoalteromonas / Q000201:enzymology* / Q000235:genetics; D013431:Sulfates / Q000378:metabolism; D013696:Temperature
chemical_list: D013431:Sulfates; D000362:Agar; D001192:Arylsulfatases
keywords: Characterization; Error-prone PCR; Mutant arylsulfatase; Thermostability improvement