title: Purification of a new galactanase from Penicillium oxalicum catalysing the hydrolysis of beta-(1----5)-galactofuran linkages.
abstract: An endo beta-(1----5)-galactofuranase from Penicillium oxalicum has been purified 91-fold. The enzyme is a basic glycoprotein with a pI 7.9 and 20% (w/w) carbohydrate content, galactose being the principal sugar. The apparent Mr of the enzyme estimated by denaturing gel electrophoresis was 77,000. The optimum pH was 5.0, and the enzyme was stable over the pH range 4.0-7.5. This enzyme hydrolyses specifically (1----5)-linked beta-D-galactofuranose residues in homo- and heterogalactans, but did not hydrolyse o-nitrophenyl galactose and beta-(1----5)-galactofuranbiose. Km and Vmax. values were 1.2 mg.ml-1 and 0.55 mumol.h-1 respectively when Eupenicillium crustaceum beta-(1----5)-galactofuran was used as substrate. The enzyme showed high affinity for different separation gels and proteins. The enzyme specificity and its mode of action showed that it could be an useful tool for analysing the fine structure of polysaccharides.
authors.affiliation: Departamento de Microbiología Aplicada, Centro de Investigaciones Biológicas (C.S.I.C.), Madrid, Spain.
authors.forename: F
authors.initials: F
authors.lastname: Reyes
authors.forename: C
authors.initials: C
authors.lastname: Alfonso
authors.forename: M J
authors.initials: MJ
authors.lastname: Martinez
authors.forename: A
authors.initials: A
authors.lastname: Prieto
authors.forename: F
authors.initials: F
authors.lastname: Santamaria
authors.forename: J A
authors.initials: JA
authors.lastname: Leal
mesh_terms: D002236:Carbohydrate Conformation; D002241:Carbohydrates / Q000032:analysis; D002850:Chromatography, Gel; D004795:Enzyme Stability; D005685:Galactans / Q000378:metabolism; D005690:Galactose / Q000032:analysis; D006026:Glycoside Hydrolases; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007526:Isoelectric Point; D008970:Molecular Weight; D010407:Penicillium / Q000201:enzymology*; D013379:Substrate Specificity; D001616:beta-Galactosidase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D002241:Carbohydrates; D005685:Galactans; D006026:Glycoside Hydrolases; C073042:endo beta-(1-5)-galactofuranase; D001616:beta-Galactosidase; D005690:Galactose