title: Characterization of a new alginate lyase from newly isolated Flavobacterium sp. S20.
abstract: Alginate lyase is a promising biocatalyst because of its application in saccharification of alginate for the production of biochemicals and renewable biofuels. This study described the isolation of a new alginate metabolizing bacterium, Flavobacterium sp. S20, from sludge samples and the characterization of its alginate lyase Alg2A. The alginate lyase gene, alg2A, was obtained by constructing and screening the genomic library of the strain S20 and overexpressed in Escherichia coli. Substrate specificity assays indicated Alg2A preferred poly-α-L-guluronate as a substrate over poly-β-D-mannuronate. In the saccharification process of a high content (10 %, w/v) of sodium alginate, the recombinant alginate lyase Alg2A yielded 152 of mM the reducing sugars after 69 h of reaction, and the amounts of oligosaccharides with a different degree of polymerization (DP) generated by Alg2A gradually accumulated without significant variation in the distribution of oligosaccharide compositions. These results indicated that Alg2A possessed high enzymatic capability for saccharifying the alginate, which could be used in saccharifying the alginate biomass prior to the main fermentation process for biofuels. In addition, Alg2A had a different endolytic reaction mode from both the two commercial alginate lyases and other alginate lyases from polysaccharide lyase family 7 owing to high yields of penta-, hex-, and hepta-saccharides in the hydrolysis products of Alg2A. Thus, Alg2A could be a good tool for the large-scale preparation of alginate oligosaccharides with high DP.
authors.affiliation: Natural Products and Glyco-Biotechnology Research Group, Liaoning Provincial Key Laboratory of Carbohydrates, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, CAS, Dalian 116023, People's Republic of China.
authors.forename: Lishuxin
authors.initials: L
authors.lastname: Huang
authors.forename: Jungang
authors.initials: J
authors.lastname: Zhou
authors.forename: Xiao
authors.initials: X
authors.lastname: Li
authors.forename: Qiang
authors.initials: Q
authors.lastname: Peng
authors.forename: Hong
authors.initials: H
authors.lastname: Lu
authors.forename: Yuguang
authors.initials: Y
authors.lastname: Du
mesh_terms: D000464:Alginates / Q000378:metabolism; D005417:Flavobacterium / Q000145:classification / Q000201:enzymology* / Q000302:isolation & purification; D020723:Glucuronic Acid / Q000378:metabolism; D006603:Hexuronic Acids / Q000378:metabolism; D009844:Oligosaccharides / Q000378:metabolism; D011133:Polysaccharide-Lyases / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D009844:Oligosaccharides; D020723:Glucuronic Acid; D011133:Polysaccharide-Lyases; C016158:poly(beta-D-mannuronate) lyase