Cloning, overexpression and characterization of a new oligoalginate lyase from a marine bacterium, Shewanella sp

• Published in: Biotechnology letters Vol. 37; no. 3; pp. 665 - 671
• Main Authors: Wang, Linna, Li, Shangyong, Yu, Wengong, Gong, Qianhong
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer-Verlag 01.03.2015; Springer Netherlands; Springer Nature B.V
• Subjects: alginate lyase; Alginates; Alginates - metabolism; Amino acids; Applied Microbiology; Bacteria; Biochemistry; Biodiesel fuels; Biofuels; Biomedical and Life Sciences; Biotechnology; Cloning; Cloning, Molecular; DNA, Bacterial - chemistry; DNA, Bacterial - genetics; DNA, Ribosomal - chemistry; DNA, Ribosomal - genetics; E coli; Electrophoresis, Polyacrylamide Gel; Encoding; Enzymatic activity; enzyme activity; Enzyme Stability; Enzymes; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Gene Expression; Genes; Glucuronic Acid - metabolism; Hexuronic Acids - metabolism; Hydrogen-Ion Concentration; Kinetics; Life Sciences; Magnesium; Microbiology; Molecular Sequence Data; Molecular Weight; Original Research Paper; Peptides; polymerase chain reaction; Polysaccharide-Lyases - chemistry; Polysaccharide-Lyases - genetics; Polysaccharide-Lyases - isolation & purification; Polysaccharide-Lyases - metabolism; Protein Sorting Signals; Recombinant; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; RNA, Ribosomal, 16S - genetics; saccharification; Sequence Analysis, DNA; Shewanella; Shewanella - enzymology; Shewanella - genetics; signal peptide; sugars; Temperature; Monomeric sugar acid; Oligoalginate lyase OalS17; Biofuel; sp. Kz7; Cloning
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-014-1706-z

PURPOSE OF WORK: 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⁻¹toward alginate, 24 U mg⁻¹toward polyM and 5 U mg⁻¹toward 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.