Purification and characterization of an alginate lyase from marine Bacterium Vibrio sp. mutant strain 510-64.
Marine Vibrio sp. 510 was chosen as a parent strain for screening high producers of alginate lyase using the complex mutagenesis of Ethyl Methanesulphonate and UV radiation treatments. The mutant strain Vibrio sp. 510-64 was selected and its alginate lyase activity was increased by 3.87-fold (reaching 46.12 EU/mg) over that of the parent strain. An extracellular alginate lyase was purified from Vibrio sp. 510-64 cultural supernatant by successive fractionation on DEAE Sepharose FF and two steps of Superdex 75. The purified enzyme yielded a single band on SDS-PAGE with the molecular weight of 34.6 kDa. Data of the N-terminal amino acid sequence indicated that this protein might be a novel alginate lyase. The substrate specificity results demonstrated that the alginate lyase had the specificity for poly G block.
Hu
Xiaoke
X
Institute of Marine Drug and Food, Ocean University of China, Qingdao, 266003, P.R. China. huxiaoke@hotmail.com
Jiang
Xiaolu
X
Hwang
Huey-Min
HM
D000595:Amino Acid Sequence; D005020:Ethyl Methanesulfonate; D008970:Molecular Weight; D016296:Mutagenesis; D011133:Polysaccharide-Lyases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D014466:Ultraviolet Rays; D044648:Undaria / Q000382:microbiology; D014733:Vibrio / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D014871:Water Microbiology
D005020:Ethyl Methanesulfonate; D011133:Polysaccharide-Lyases; C016158:poly(beta-D-mannuronate) lyase