title: Purification and Characterization of Hyaluronate Lyase from Arthrobacter globiformis A152.
abstract: A hyaluronate lyase was obtained by cultivating Arthrobacter globiformis strain A152. The enzyme was purified to homogeneity from the supernatant by ammonium sulfate fractionation, Q Sepharose Fast Flow, and Sephadex G-100 chromatography. The purification resulted in a 32.78-fold increase in hyaluronate lyase activity with specific activity of 297.2 U/mg. The molecular weight of the enzyme determined by SDS-PAGE was approximately 73.7 kDa. Using hyaluronic acid (HA) as a substrate, the maximal reaction rate (Vmax) and the Michaelis-Menten constant (Km) of hyaluronate lyase were found to be 4.76 μmol/min/ml and 0.11 mg/ml, respectively. The optimum pH and temperature values for hyaluronate lyase activity were pH 6.0 and 42 °C, respectively. This enzyme was stable at pH 4-10, 5-7, and 5-7 at 4, 37, and 42 °C, respectively. Investigation about temperature effects on hyaluronate lyase displayed that it was stable at 30-37 °C and also showed high activity at 37 °C. The enzymatic activity was enhanced by Ca2+and was strongly inhibited by Cu2+and SDS. These properties suggested that the hyaluronate lyase in this study could bring promising prospects in medical and industry applications.
authors.affiliation: School of Medicine and Pharmacy, Ocean University of China, Qingdao, 266003, People's Republic of China.
authors.forename: Changliang
authors.initials: C
authors.lastname: Zhu
authors.affiliation: School of Medicine and Pharmacy, Ocean University of China, Qingdao, 266003, People's Republic of China.|Marine Biomedical Research Institute of Qingdao, 23 Hongkong East Road, Qingdao, 266071, People's Republic of China.
authors.forename: Jingliang
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Medical School, Huanghe Science and Technology College, Zhengzhou, 450063, People's Republic of China.
authors.forename: Liyan
authors.initials: L
authors.lastname: Li
authors.affiliation: College of Food Science and Engineering, Ocean University of China, 5 Yushan Road, Qingdao, 266003, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.affiliation: College of Food Science and Engineering, Ocean University of China, 5 Yushan Road, Qingdao, 266003, People's Republic of China.
authors.forename: Yanhui
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: School of Medicine and Pharmacy, Ocean University of China, Qingdao, 266003, People's Republic of China.|Marine Biomedical Research Institute of Qingdao, 23 Hongkong East Road, Qingdao, 266071, People's Republic of China.
authors.forename: Zhaopeng
authors.initials: Z
authors.lastname: Shen
authors.affiliation: School of Medicine and Pharmacy, Ocean University of China, Qingdao, 266003, People's Republic of China.
authors.forename: Huashi
authors.initials: H
authors.lastname: Guan
authors.affiliation: College of Food Science and Engineering, Ocean University of China, 5 Yushan Road, Qingdao, 266003, People's Republic of China. jiangxl@ouc.edu.cn.
authors.forename: Xiaolu
authors.initials: X
authors.lastname: Jiang
mesh_terms: D001173:Arthrobacter / Q000737:chemistry* / Q000201:enzymology; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D002118:Calcium / Q000737:chemistry; D002413:Cations, Divalent; D003300:Copper / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006820:Hyaluronic Acid / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D011133:Polysaccharide-Lyases / Q000737:chemistry* / Q000302:isolation & purification; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D002413:Cations, Divalent; D012967:Sodium Dodecyl Sulfate; D003300:Copper; D006820:Hyaluronic Acid; D011133:Polysaccharide-Lyases; C100002:hyaluronate lyase; D002118:Calcium
keywords: Biocatalyst; Characterization; Hyaluronate lyase; Purification