Catalytic properties and mode of action of endo-(1-->3)-beta-D-glucanase and beta-D-glucosidase from the marine mollusk Littorina kurila

Carbohydr Res. 2008 Sep 22;343(14):2393-400. doi: 10.1016/j.carres.2008.06.025. Epub 2008 Jul 6.

Abstract

A complex of the enzymes from the liver of the marine mollusk Littorina kurila that hydrolyzes laminaran was investigated. Two (1-->3)-beta-d-glucanases (G-I and G-II) were isolated. The molecular mass of G-I as estimated by gel-permeation chromatography and SDS-PAGE analysis was 32 and 40kDa, respectively. The G-II molecular mass according to SDS-PAGE analysis was about 200kDa. The pH optimum for both G-I and G-II was pH 5.4. The G-I had narrow substrate specificity and hydrolyzed only the (1-->3)-beta-d-glucosidic bonds in the mixed (1-->3),(1-->6)- and (1-->3),(1-->4)-beta-d-glucans down to glucose and glucooligosaccharides. This enzyme acted with retention of the anomeric configuration and catalyzed a transglycosylation reaction. G-I was classified as the glucan endo-(1-->3)-beta-d-glucosidase (EC 3.2.1.39). G-II exhibited both exo-glucanase and beta-d-glucoside activities. This enzyme released from the laminaran glucose as a single product, but retained the anomeric center configuration and possessed transglycosylation activity. The hydrolysis rate of glucooligosaccharides by G-I decreased with an increase of the substrate's degree of polymerization. In addition to (1-->3)-beta-d-glucanase activity, the enzyme had the ability to hydrolyze p-nitrophenyl beta-d-glucoside and beta-d-glucobioses: laminaribiose, gentiobiose, and cellobiose, with the rate ratio of 50:12:1. G-II may correspond to beta-d-glucoside glucohydrolase (EC 3.2.1.21).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Endo-1,3(4)-beta-Glucanase / chemistry
  • Endo-1,3(4)-beta-Glucanase / isolation & purification
  • Endo-1,3(4)-beta-Glucanase / metabolism*
  • Glucans
  • Glucosidases / chemistry
  • Glucosidases / isolation & purification
  • Glucosidases / metabolism*
  • Hydrogen-Ion Concentration
  • Hydrolysis
  • Liver / enzymology
  • Marine Biology
  • Molecular Weight
  • Nuclear Magnetic Resonance, Biomolecular
  • Polysaccharides / metabolism
  • Snails / enzymology*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Substrate Specificity

Substances

  • Glucans
  • Polysaccharides
  • laminaran
  • Glucosidases
  • Endo-1,3(4)-beta-Glucanase