title: Studies on the chromatographic fractionation of Trichoderma reesei cellulases by hydrophobic interaction.
abstract: This work reports new studies on cellulases fractionation by hydrophobic interaction chromatography. The purification procedure for the Trichoderma reesei cellulase complex consists of gel permeation chromatography on Sephadex G-25M followed by an ultrafiltration step. The concentrated enzyme solution was then fractionated on Sepharose CL-6B modified by covalent immobilization of 1,4-butanediol diglycidyl ether. The influence of the mobile phase composition on the chromatographic behaviour of the T. reesei cellulase complex was investigated. By using 13% (w/v) ammonium sulphate in eluent buffer, a selective separation of beta-glucosidase with a two-fold increase in specific activity and a recovery of 60% cellobiase activity were obtained. Other commercial hydrophobic supports (octyl- and phenyl-Sepharose) were also tested and compared under the same conditions.
authors.affiliation: Departamento de Química, Universidade da Beira Interior, Covilhã, Portugal.
authors.forename: C T
authors.initials: CT
authors.lastname: Tomaz
authors.forename: J A
authors.initials: JA
authors.lastname: Queiroz
mesh_terms: D002480:Cellulase / Q000302:isolation & purification*; D005591:Chemical Fractionation; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002845:Chromatography / Q000379:methods*; D002850:Chromatography, Gel; D014242:Trichoderma / Q000201:enzymology*; D001617:beta-Glucosidase / Q000302:isolation & purification
chemical_list: D001617:beta-Glucosidase; D002480:Cellulase