Purification, characterization and synergism in autolysis of a group of 1,3-ß-glucan hydrolases from the pilei of Coprinopsis cinerea fruiting bodies.
Microbiology (Reading)
; 161(10): 1978-1989, 2015 Oct.
Article
em En
| MEDLINE
| ID: mdl-26199012
ABSTRACT
Using a combined chromatography method, we simultaneously purified three protein fractions (II-2, II-3 and II-4) with 1,3-ß-glucanase activity from extraction of pilei of Coprinopsis cinerea fruiting bodies. MALDI-TOF/TOF amino acid sequencing showed that these three fractions matched a putative exo-1,3-ß-glucanase, a putative glucan 1,3-ß-glucosidase and a putative glycosyl hydrolase family 16 protein annotated in the C. cinerea genome, respectively; however, they were characterized as a 1,3-ß-glucosidase, an exo-1,3-ß-glucanase and an endo-1,3-ß-glucanase, respectively, by analysis of their substrate specificities and modes of action. This study explored how these three 1,3-ß-glucoside hydrolases synergistically acted on laminarin the endo-1,3-ß-glucanase hydrolysed internal glycosidic bonds of laminarin to generate 1,3-ß-oligosaccharides of various lengths, the exo-1,3-ß-glucanase cleaved the longer-chain laminarioligosaccharides into short-chain disaccharides, laminaribiose and gentiobiose, and the 1,3-ß-glucosidase further hydrolysed laminaribiose to glucose. The remaining gentiobiose must be hydrolysed by other 1,6-ß-glucosidases. Therefore, the endo-1,3-ß-glucanase, exo-1,3-ß-glucanase and 1,3-ß-glucosidase may act synergistically to completely degrade the 1,3-ß-glucan backbone of the C. cinerea cell wall during fruiting body autolysis. These three 1,3-ß-glucoside hydrolases share a similar optimum pH and optimum temperature, supporting the speculation that these enzymes work together under the same conditions to degrade 1,3-ß-glucan in the C. cinerea cell wall during fruiting body autolysis.
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Base de dados:
MEDLINE
Assunto principal:
Agaricales
/
Glucana 1,3-beta-Glucosidase
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Glucanos
Idioma:
En
Ano de publicação:
2015
Tipo de documento:
Article