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.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Alginates / metabolism*
-
Cloning, Molecular
-
DNA, Bacterial / chemistry
-
DNA, Bacterial / genetics
-
DNA, Ribosomal / chemistry
-
DNA, Ribosomal / genetics
-
Electrophoresis, Polyacrylamide Gel
-
Enzyme Stability
-
Escherichia coli / genetics
-
Escherichia coli / metabolism
-
Gene Expression
-
Glucuronic Acid / metabolism
-
Hexuronic Acids / metabolism
-
Hydrogen-Ion Concentration
-
Kinetics
-
Molecular Sequence Data
-
Molecular Weight
-
Polysaccharide-Lyases / chemistry
-
Polysaccharide-Lyases / genetics
-
Polysaccharide-Lyases / isolation & purification
-
Polysaccharide-Lyases / metabolism*
-
Protein Sorting Signals
-
RNA, Ribosomal, 16S / genetics
-
Recombinant Proteins / chemistry
-
Recombinant Proteins / genetics
-
Recombinant Proteins / isolation & purification
-
Recombinant Proteins / metabolism
-
Sequence Analysis, DNA
-
Shewanella / enzymology*
-
Shewanella / genetics
-
Temperature
Substances
-
Alginates
-
DNA, Bacterial
-
DNA, Ribosomal
-
Hexuronic Acids
-
Protein Sorting Signals
-
RNA, Ribosomal, 16S
-
Recombinant Proteins
-
Glucuronic Acid
-
Polysaccharide-Lyases
-
poly(beta-D-mannuronate) lyase
Associated data
-
GENBANK/KF673475
-
GENBANK/KJ094505