Purification and characterisation of a bifunctional alginate lyase from novel Isoptericola halotolerans CGMCC 5336.
A novel halophilic alginate-degrading microorganism was isolated from rotten seaweed and identified as Isoptericola halotolerans CGMCC5336. The lyase from the strain was purified to homogeneity by combining of ammonium sulfate fractionation and anion-exchange chromatography with a specific activity of 8409.19 U/ml and a recovery of 25.07%. This enzyme was a monomer with a molecular mass of approximately 28 kDa. The optimal temperature and pH were 50 °C and pH 7.0, respectively. The lyase maintained stability at neutral pH (7.0-8.0) and temperatures below 50 °C. Metal ions including Na(+), Mg(2+), Mn(2+), and Ca(2+) notably increased the activity of the enzyme. With sodium alginate as the substrate, the Km and Vmax were 0.26 mg/ml and 1.31 mg/ml min, respectively. The alginate lyase had substrate specificity for polyguluronate and polymannuronate units in alginate molecules, indicating its bifunctionality. These excellent characteristics demonstrated the potential applications in alginate oligosaccharides production with low polymerisation degrees.
School of Pharmaceutical Science, Jiangnan University, Wuxi 214122, PR China.
Wenfang
W
Dou
Dan
D
Wei
Hui
H
Li
Heng
H
Li
Muhammad Masfiqur
MM
Rahman
Jinsong
J
Shi
Zhenghong
Z
Xu
Yanhe
Y
Ma
D000192:Actinomycetales / Q000737:chemistry / Q000201:enzymology*; D000464:Alginates / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D002118:Calcium / Q000737:chemistry; D002413:Cations, Divalent; D002414:Cations, Monovalent; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000737:chemistry; D008345:Manganese / Q000737:chemistry; D008970:Molecular Weight; D011133:Polysaccharide-Lyases / Q000737:chemistry* / Q000302:isolation & purification; D011135:Polysaccharides, Bacterial / Q000737:chemistry; D012624:Seaweed / Q000382:microbiology; D012964:Sodium / Q000737:chemistry; D013379:Substrate Specificity; D013696:Temperature
D000464:Alginates; D001426:Bacterial Proteins; D002413:Cations, Divalent; D002414:Cations, Monovalent; D006603:Hexuronic Acids; D011135:Polysaccharides, Bacterial; C111091:polyguluronic acid; D008345:Manganese; D020723:Glucuronic Acid; D012964:Sodium; D011133:Polysaccharide-Lyases; C016158:poly(beta-D-mannuronate) lyase; D008274:Magnesium; D002118:Calcium
Alginate lyase; Bifunctional; Characterisation; Isoptericola halotolerans; Purification