Purification and characterization of a new kappa-carrageenase from a marine Cytophaga-like bacterium

Eur J Biochem. 1991 Oct 1;201(1):241-7. doi: 10.1111/j.1432-1033.1991.tb16280.x.

Abstract

A bacterial strain able to degrade various sulfated galactans (carrageenans and agar) was isolated from the marine red alga Delesseria sanguinea. From the cell-free supernatant of cultures grown on crude lambda-carrageenan, a kappa-carrageenase was purified by ammonium sulfate fractionation, gel filtration on Sephacryl S 200 HR and ion-exchange chromatography on DEAE--Sepharose-CL6B. The purified kappa-carrageenase was detected as a single protein upon SDS/PAGE. Its molecular mass was estimated at 40 kDa. Activity was observed against kappa-carrageenan over the pH range 5.0-8.5 and was optimal at pH 7.2 in Tris buffer or 7.0 in Mops buffer. The enzyme activity remained stable at 30 degrees C, but only for up to 1 h at 40 degrees C. Analysis of the degradation products of the kappa-carrageenase by gel filtration and 13C-NMR spectroscopy indicated that the enzyme degrades kappa-carrageenan down to the level of the kappa-neocarratetraose sulfate. The properties of this new enzyme are compared with those of previously characterized carrageenases.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins*
  • Carrageenan / metabolism
  • Chromatography
  • Glycoside Hydrolases / isolation & purification*
  • Glycoside Hydrolases / metabolism
  • Hydrolysis
  • Kinetics
  • Magnetic Resonance Spectroscopy
  • Molecular Weight
  • Rhodophyta / enzymology*
  • Substrate Specificity

Substances

  • Bacterial Proteins
  • Carrageenan
  • Glycoside Hydrolases
  • kappa-carrageenase protein, Alteromonadaceae