title: Purification and characterization of polysaccharides degradases produced by Alteromonas sp. A321.
abstract: Two different degradases from Alteromonas sp. A321 for polysaccharides from Enteromorpha prolifera (DPE-L and DPE-P) were purified to homogeneity. The molecular weights of purified DPE-L and DPE-P were 75.2 and 102.5 kDa, respectively, and their internal sequences were analysed by LC-MS-MS. The enzymes exhibited an optimum temperature of 30-40 °C (DPE-L) and 35-45 °C (DPE-P), an optimum pH of 7.0 (DPE-L) and 6.0 (DPE-P). DPE-P was highly stable in the presence of EDTA and 1,10-phenanthroline while DPE-L was inhibited by 1,10-phenanthroline. The Km values of DPE-L and DPE-P were 2.93 mg/ml and 0.31 mg/ml and the Vmax values were 6.11 μmol/min/ml and 2.88 μmol/min/ml, respectively. Results of HPLC and ESI-MS analyses showed that enzymatic products were: Rha1(SO3H)1, Rha1(SO3H)1Gluc1, Rha2(SO3H)2Gluc1, and Rha3(SO3H)3Gluc1Xyl1 by DPE-L, and Glu2, Glu3, plus Glu4 by DPE-P, respectively. Thus DPE-L and DPE-P can be used to produce oligosaccharides which potentially revealed more of structure of polysaccharides from E. prolifera.
authors.affiliation: School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, PR China.
authors.forename: Yinping
authors.initials: Y
authors.lastname: Li
authors.affiliation: Qingdao University, Qingdao 266071, PR China.
authors.forename: Weihua
authors.initials: W
authors.lastname: Li
authors.affiliation: College of Food Science and Engineering, Ocean University of China, Qingdao 266003, PR China.
authors.forename: Gaoli
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Jinan Fruit Research Institute, All China Federation of Supply and Marketing Co-operatives, Jinan 250014, PR China.
authors.forename: Xuqiang
authors.initials: X
authors.lastname: Lü
authors.affiliation: Biology Department, Jackson State University, Jackson, Mississippi 39217, USA.
authors.forename: Hueymin
authors.initials: H
authors.lastname: Hwang
authors.affiliation: Biology Department, Jackson State University, Jackson, Mississippi 39217, USA.
authors.forename: Winfred G
authors.initials: WG
authors.lastname: Aker
authors.affiliation: School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, PR China.
authors.forename: Huashi
authors.initials: H
authors.lastname: Guan
authors.affiliation: College of Food Science and Engineering, Ocean University of China, Qingdao 266003, PR China. Electronic address: pengwang@ouc.edu.cn.
authors.forename: Peng
authors.initials: P
authors.lastname: Wang
mesh_terms: D020591:Alteromonas / Q000201:enzymology* / Q000378:metabolism; D000595:Amino Acid Sequence; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D008670:Metals / Q000494:pharmacology; D008970:Molecular Weight; D011134:Polysaccharides / Q000378:metabolism*; D013501:Surface-Active Agents / Q000494:pharmacology; D013696:Temperature; D001617:beta-Glucosidase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D008670:Metals; D011134:Polysaccharides; D013501:Surface-Active Agents; D001617:beta-Glucosidase
keywords: Characterization; Degradase for polysaccharides from Enteromorpha prolifera; Sulfate oligorhamnose