Malays. Appl. Biol. (2017) 46(2): 11–19
CHARACTERISATION OF RECOMBINANT Trichoderma reesei
CELLOBIOHYDROLASE AND THE POTENTIAL OF
CELLULASE MIXTURE IN HYDROLYZING OIL PALM
EMPTY FRUIT BUNCHES
DORIS QUAY HUAI XIA1, YONG HUI YEE1, ROSLI MD ILLIAS2, NOR MUHAMMAD MAHADI3,
FARAH DIBA ABU BAKAR1 and ABDUL MUNIR ABDUL MURAD1*
1School of Biosciences and Biotechnology, Faculty of Science and Technology,
Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor, Malaysia
2Department of Bioprocess Engineering, Faculty of Chemical Engineering, Universiti Teknologi Malaysia,
81310 Skudai, Johor, Malaysia
3Malaysia Genome Institute, Jalan Bangi, 43000 Kajang, Selangor, Malaysia of Agriculture Science and Biotechnology, Faculty of Bioresource and Food Industry, Universiti Sultan Zainal Abidin, Kampus Besut, 22200 Besut, Terengganu, Malaysia
*Email: This e-mail address is being protected from spambots. You need JavaScript enabled to view it / This e-mail address is being protected from spambots. You need JavaScript enabled to view it
Accepted 2 January 2017, Published online 27 June 2017
ABSTRACT
The potential for recombinant cellulases from fungi to hydrolyze oil palm empty fruit bunches (OPEFB) into simple sugars was investigated. A mixture of two recombinant enzymes consisting of a cellobiohydrolase from Trichoderma reesei and an endoglucanase from Aspergillus niger, was evaluated for OPEFB hydrolysis. The development of A. niger endoglucanase has been described previously. In this work, the development of a recombinant cellobiohydrolase and the activity of this enzyme mixture towards OPEFB hydrolysis were described. To obtain recombinant cellobiohydrolase, the cDNA encoding for cellobiohydrolase, cbhII, was isolated from the T. reesei strain M5, and the gene was expressed in the methylotrophic yeast, Pichia pastoris. Partially purified CbhII demonstrated optimum activity at 50°C and pH 5.0. This enzyme was shown to hydrolyze Avicel at a concentration of 0.486 U/mg under optimum conditions. Hydrolysis of pretreated OPEFB using CbhII and endoglucanase EglA showed that the total reducing sugar produced was higher in the enzyme mixture reaction than were produced in an individual enzyme reaction. Addition of ?-glucosidase and an enzyme cofactor significantly increased enzyme activity and OPEFB cellulose hydrolysis. This study demonstrated the capability of an enzyme mixture to produce sugars from oil palm lignocellulosic waste.
Key words: Cellobiohydrolase, Trichoderma reesei, enzyme mixture, oil palm empty fruit bunch
