Characterization of a new endo-type alginate lyase from Vibrio sp. W13

Int J Biol Macromol. 2015 Apr:75:330-7. doi: 10.1016/j.ijbiomac.2015.01.053. Epub 2015 Feb 7.

Abstract

A gene, encoding a new alginate lyase Algb, was identified and cloned from marine bacterium Vibrio sp. W13. The recombinant alginate lyase was characterized followed by being purified on Ni-NTA Sepharose. It exhibited the highest activity (457 U/mg) at pH 8.0 and 30 °C. Interestingly, Algb possessed broader substrate specificity. It showed activities toward both polyM (poly β-D-mannuronate) and polyG (poly α-L-guluronate). Furthermore, K(m) values of Algb toward alginate (0.67 mg/ml) and polyMG (0.50 mg/ml) are lower than those toward polyG (1.04 mg/ml) and polyM (6.90 mg/ml). The TLC and ESI-MS analysis suggested that Algb mainly released oligosaccharides with DP of 2-5 from the four kinds of substrates in an endolytic manner. Therefore, it may be a potent tool to produce alginate oligosaccharides with low DP.

Keywords: Alginate lyase; Polysaccharide lyase family 7; Vibrio sp..

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Chromatography, Thin Layer
  • Cloning, Molecular
  • Electrophoresis, Polyacrylamide Gel
  • Ions
  • Kinetics
  • Metals / pharmacology
  • Molecular Sequence Data
  • Polysaccharide-Lyases / chemistry
  • Polysaccharide-Lyases / genetics
  • Polysaccharide-Lyases / isolation & purification
  • Polysaccharide-Lyases / metabolism*
  • Recombinant Proteins / isolation & purification
  • Sequence Analysis, Protein
  • Sodium Chloride / pharmacology
  • Spectrometry, Mass, Electrospray Ionization
  • Substrate Specificity / drug effects
  • Vibrio / enzymology*

Substances

  • Ions
  • Metals
  • Recombinant Proteins
  • Sodium Chloride
  • Polysaccharide-Lyases
  • poly(beta-D-mannuronate) lyase