title: Purification and characterization of chondroitinase ABC from Acinetobacter sp. C26.
abstract: An extracellular chondroitinase ABC (ChSase ABC, EC 4.2.2.4) produced by cultivating Acinetobacter sp. C26, was purified to homogeneity from the supernatant by ammonium sulfate fractionation, Q-Sepharose Fast Flow and Sephadex G-100 chromatography. The 76kDa enzyme was purified 48.09-fold to homogeneity with specific activity of 348.64U/mg, Using the chondroitin sulfate A (CS-A) as substrate, the maximal reaction rate (Vmax) and Michaelis-Menten constant (Km) of ChSase ABC were found to be 10.471μmol/min/ml and 0.105mg/ml, respectively. The enzyme showed the highest activity at the optimal conditions of pH 6.0 and 42 ∘C, respectively. This enzyme was stable at pH 5-10, 5-9 and 5-7 at 4°C, 37°C and 42°C, respectively. Investigation about thermal stability of ChSase ABC displayed that it was stable at 37°C. ChSase ABC activity was increased in presence of Na+, K+, Mn2+, 1,10-phenanthrolin and strongly inhibited by Cu2+, Hg2+, Al3+and SDS. These properties suggested that ChSase ABC from Acinetobacter sp. C26 bring promising prospects in medical and industry applications.
authors: School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao 266003, PR China. Electronic address: chlzhu@163.com.
authors: Changliang
authors: C
authors: Zhu
authors: School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao 266003, PR China; Marine Biomedical Research Institute of Qingdao, 23 Hongkong East Road, Qingdao 266071, PR China. Electronic address: jlzhang1985@163.com.
authors: Jingliang
authors: J
authors: Zhang
authors: College of Food Science and Engineering, Ocean University of China, 5 Yushan Road, Qingdao 266003, PR China. Electronic address: zjys1991@163.com.
authors: Jing
authors: J
authors: Zhang
authors: College of Food Science and Engineering, Ocean University of China, 5 Yushan Road, Qingdao 266003, PR China. Electronic address: shannongjyh@163.com.
authors: Yanhui
authors: Y
authors: Jiang
authors: School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao 266003, PR China; Marine Biomedical Research Institute of Qingdao, 23 Hongkong East Road, Qingdao 266071, PR China. Electronic address: shenzhaopeng1987@163.com.
authors: Zhaopeng
authors: Z
authors: Shen
authors: School of Medicine and Pharmacy, Ocean University of China, 5 Yushan Road, Qingdao 266003, PR China. Electronic address: sunyang@ouc.edu.cn.
authors: Huashi
authors: H
authors: Guan
authors: College of Food Science and Engineering, Ocean University of China, 5 Yushan Road, Qingdao 266003, PR China. Electronic address: jiangxl@ouc.edu.cn.
authors: Xiaolu
authors: X
authors: Jiang
mesh_terms: D000150:Acinetobacter / Q000201:enzymology*; D000595:Amino Acid Sequence; D000645:Ammonium Sulfate / Q000378:metabolism; D019765:Chondroitin ABC Lyase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D008670:Metals / Q000494:pharmacology; D013501:Surface-Active Agents / Q000494:pharmacology; D013696:Temperature
chemical_list: D004791:Enzyme Inhibitors; D008670:Metals; D013501:Surface-Active Agents; D019765:Chondroitin ABC Lyase; D000645:Ammonium Sulfate
keywords: Acinetobacter; Characterization; Chondroitinase ABC; Purification