title: Cloning, overexpression and characterization of a new oligoalginate lyase from a marine bacterium, Shewanella sp.
abstract: Is to report an oligoalginate lyase with high enzymatic activity and high-level expression. Using site-finding PCR and degenerate PCR, a gene (designated oalS17) encoding a new oligoalginate lyase was cloned from Shewanella sp. Kz7 and expressed in Escherichia coli. The gene consisted of 2,292 bp with deduced amino acid size of 763 including a putative signal peptide of 44 amino acid residues belonging to polysaccharide lyase (PL) family 17. The recombinant protein was most active at 50 °C and pH 6.2 in 50 mM phosphate buffer. It degraded alginate more efficiently than polyM and polyG block into a monomeric sugar acid, with a specific activity of 32 U mg(-1) toward alginate, 24 U mg(-1) toward polyM and 5 U mg(-1) toward polyG. With the high-level expression and high enzymatic activity, the recombinant oligoalginate lyase OalS17 could be a potential enzyme for further research on alginate saccharification and biofuels production.
authors.affiliation: Key Laboratory of Marine Drugs, Chinese Ministry of Education; Shandong Provincial Key Laboratory of Glycoscience & Glycotechnology; School of Medicine and Pharmacy, Ocean University of China, Qingdao, 266003, People's Republic of China, wlnwfllsy@163.com.
authors.forename: Linna
authors.initials: L
authors.lastname: Wang
authors.forename: Shangyong
authors.initials: S
authors.lastname: Li
authors.forename: Wengong
authors.initials: W
authors.lastname: Yu
authors.forename: Qianhong
authors.initials: Q
authors.lastname: Gong
mesh_terms: D000464:Alginates / Q000378:metabolism*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D020723:Glucuronic Acid / Q000378:metabolism; D006603:Hexuronic Acids / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011133:Polysaccharide-Lyases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D021382:Protein Sorting Signals; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D020592:Shewanella / Q000201:enzymology* / Q000235:genetics; D013696:Temperature
chemical_list: D000464:Alginates; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D006603:Hexuronic Acids; D021382:Protein Sorting Signals; D012336:RNA, Ribosomal, 16S; D011994:Recombinant Proteins; D020723:Glucuronic Acid; D011133:Polysaccharide-Lyases; C016158:poly(beta-D-mannuronate) lyase