Comparative study on the fractionation of cellulases on some hydrophobic interaction chromatography adsorbents.
In this work, a comparative study for the fractionation of Trichoderma reesei cellulases on five different hydrophobic interaction chromatography adsorbents (Butyl-Sepharose 4 FF, Phenyl-Sepharose 6 FF, Octyl-Sepharose 4 FF, Epoxy-Sepharose CL-6B and Polypropylene glycol-Sepharose CL-6B) is shown. The influence of the mobile phase composition on the chromatographic behaviour of T. reesei cellulases complex was evaluated using different concentrations of ammonium sulphate in the eluent buffer. A selective separation of beta-glucosidase with two-fold increase in specific activity and good recoveries of cellobiase activity were obtained with Butyl-Sepharose 4 FF and Phenyl-Sepharose 6 FF using 7% (w/v) ammonium sulphate in the eluent buffer. A beta-glucosidase fractionation was also obtained with Epoxy-Sepharose CL-6B, using 13% (w/v) of the salt in the mobile phase.
Tomaz
Candida T
CT
Departamento de Química, Universidade do Beira Interior, Covilhã, Portugal.
Duarte
Dina
D
Queiroz
João A
JA
D002480:Cellulase / Q000302:isolation & purification*; D002853:Chromatography, Liquid / Q000379:methods*; D013056:Spectrophotometry, Ultraviolet
D002480:Cellulase