Abstract
Fractionation of proteins secreted into the culture medium by intact cells and protoplasts of Pichia polymorpha showing enzyme activity against laminarin, pustulan or p-nitrophenyl-beta-D-glucopyranoside has been performed, and the results compared with those obtained with cell-free extracts and lysed protoplasts. Fractionation with DEAE Sephadex A50 has proved to be the best method, yielding at least three fractions which hydrolyse laminarin. One of these fractions was active on both laminarin and pustulan. Filtration on Sephadex G-100 column only yielded one active preparation. Evidence supporting the conclusion that there are three different beta-glucanases located in the periplasmic space is presented.
MeSH Terms
Ascomycota/enzymology
Cell-Free System
Chromatography, DEAE-Cellulose
Chromatography, Gel
Glucan Endo-1,3-beta-D-Glucosidase/isolation & purification
Glucans
Glucose
Glycoside Hydrolases/isolation & purification
Molecular Weight
Pichia/enzymology
Polysaccharides
Protoplasts/enzymology
Chemicals
Glucans
Polysaccharides
pustulan
laminaran
Glycoside Hydrolases
Glucan Endo-1,3-beta-D-Glucosidase
Glucose
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Villa T G
Notario V
Villanueva J R
References (11)
11 references, click to expand
-
The structure of a beta-(1 leads to 3)-D-glucan from yeast cell walls.
Biochem J. 1973 Sep;135(1):19-30
PMID: 4359920
-
Isolation of glucanase-containing vesicles from budding yeast.
Arch Mikrobiol. 1972;82(3):189-205
PMID: 4337552
-
Glucosidases and exo-glucanases.
Can J Biochem. 1968 Jan;46(1):25-34
PMID: 5689394
-
Notes on sugar determination.
J Biol Chem. 1952 Mar;195(1):19-23
PMID: 14938350
-
Detection of sugars on paper chromatograms.
Nature. 1950 Sep 9;166(4219):444-5
PMID: 14775715
-
Exo-beta-glucanases in yeast.
Biochem J. 1968 Sep;109(3):347-60
PMID: 5685856
-
The structure of the yeast cell wall. Solubilization of a marker enzyme, -fructofuranosidase, by the autolytic enzyme system.
J Biol Chem. 1972 Feb 25;247(4):1161-9
PMID: 4551513
-
Extracellular beta-glucanases of the yeast, Saccharomyces cerevisiae.
Biochim Biophys Acta. 1973 Sep 15;321(1):246-55
PMID: 4584394
-
Protein measurement with the Folin phenol reagent.
J Biol Chem. 1951 Nov;193(1):265-75
PMID: 14907713
-
TRANSGLUCOSYLASE ACTIVITY OF FUNGAL LAMINARINASES.
Nature. 1964 May 2;202:454-6
PMID: 14167820
-
Beta-glucanase of yeast.
Biochem Biophys Res Commun. 1965 May 18;19(5):623-9
PMID: 5834714