Large-scale production of cellulose-binding domains : adsorption studies using ...

• Author(s): Pinto, João Ricardo ; Carvalho, Joana ; Mota, M. ; Gama, F. M.
• Date: 2006
• Persistent ID: http://hdl.handle.net/1822/5498
• Origin: RepositóriUM - Universidade do Minho
• Subject(s): Adsorption; Cellulose-binding domains; FITC; Proteolysis
• Document Type: Article
• Language: English

Description

A method for the gram-scale production of cellulose-binding domains (CBD) through the proteolytic digestion of a commercial nzymatic preparation (Celluclast) was developed. The CBD obtained, isolated from Trichoderma reesei cellobiohydrolase I, is highly pure and heavily glycosylated. The purified peptide has a molecular weight of 8.43 kDa, comprising the binding module, a part of the linker, and about 30% glycosidic moiety. Its properties may thus be different from recombinant ones expressed in bacteria. CBDfluorescein isothiocyanate conjugates were used to study the CBD-cellulose interaction. The presence of fluorescent peptides adsorbed on crystalline and amorphous cellulose fibers suggests that amorphous regions have a higher concentration of binding sites. The adsorption is reversible, but desorption is a very slow process.