Depolymerization of Alginate Into a Monomeric Sugar Acid Using Alg17C, an Exo-Oligoalginate Lyase Cloned From Saccharophagus Degradans 2-40

Appl Microbiol Biotechnol. 2012 Mar;93(5):2233-9. doi: 10.1007/s00253-012-3882-x. Epub 2012 Jan 27.

Abstract

Macroalgae are considered to be promising biomass for fuels and chemicals production. To utilize brown macroalgae as biomass, the degradation of alginate, which is the main carbohydrate of brown macroalgae, into monomeric units is a critical prerequisite step. Saccharophagus degradans 2-40 is capable of degrading more than ten different polysaccharides including alginate, and its genome sequence demonstrated that this bacterium contains several putative alginate lyase genes including alg17C. The gene for Alg17C, which is classified into the PL-17 family, was cloned and overexpressed in Escherichia coli. The recombinant Alg17C was found to preferentially act on oligoalginates with degrees of polymerization higher than 2 to produce the alginate monomer, 4-deoxy-L: -erythro-5-hexoseulose uronic acid. The optimal pH and temperature for Alg17C were found to be 6 and 40 °C, respectively. The K (M) and V (max) of Alg17C were 35.2 mg/ml and 41.7 U/mg, respectively. Based on the results of this study, Alg17C could be used as the key enzyme to produce alginate monomers in the process of utilizing alginate for biofuels and chemicals production.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alginates / metabolism*
  • Alteromonadaceae / enzymology*
  • Alteromonadaceae / genetics
  • Cloning, Molecular
  • Enzyme Stability
  • Escherichia coli / genetics
  • Gene Expression
  • Hydrogen-Ion Concentration
  • Kinetics
  • Polysaccharide-Lyases / chemistry
  • Polysaccharide-Lyases / genetics
  • Polysaccharide-Lyases / isolation & purification
  • Polysaccharide-Lyases / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Substrate Specificity
  • Sugar Acids / metabolism*
  • Temperature

Substances

  • Alginates
  • Recombinant Proteins
  • Sugar Acids
  • Polysaccharide-Lyases
  • poly(beta-D-mannuronate) lyase