Abstract
β-glucosidases have characteristics of biotechnological interest and have thus become important industrial enzymes.In this study, β-glucosidase produced by Trichoderma viride from cow dung was subjected to a three step purification process involving ammonium sulphate precipitation, gel filtration by Sephadex G-100 and ion exchange chromatography by DEAE-Sephadex A-25. The elution profile on Sephadex G-100 resulted in a single broad peak (fractions 9-21) which had a yield of 3.7% and a purification fold of 4.29 with a specific activity of 25.70 µmol/min/mg proteins while the elution profile on DEAE-Sephadex A-25 resulted in a single broad peak (fraction 8-14) which had a yield of 2.76% and a purification fold of 22.14 with a specific activity of 132.41µmol/min/mg of protein. The purified enzyme was obtained as a single band and had a molecular mass of 51.8 kDa on SDS-PAGE. This results provide support for further studies of this enzyme towards revealing its potential biotechnological applications.
Highlights
INTRODUCTION β-glucosidase (EC 3.2.1.21) is a component of cellulase complex and has a synergistic action on the degradation of cellulose with endoglucanase (EC 3.2.1.4) and exoglucanase (EC 3.2.1.91) (Del pozo et al, 2012)
Some of the industrial applications that employ the use β-glucosidase include biofuel production, cassava detoxification, food and feed processing (Obilie et al, 2004; Coughlan, 1985) Cellulases are currently the third largest industrial enzymes being sold at commercial level worldwide
The cellulolytic enzymes from Trichoderma reesei have been investigated thoroughly (Saloheimo et al.,1997; Arja et al.,2004), the quantity of β-glucosidase secreted by T. reesei is not a enough for effective conversion of cellulose to glucose (Sorensen et al, 2013)
Summary
INTRODUCTION β-glucosidase (EC 3.2.1.21) is a component of cellulase complex and has a synergistic action on the degradation of cellulose with endoglucanase (EC 3.2.1.4) and exoglucanase (EC 3.2.1.91) (Del pozo et al, 2012). The ability of fungi to grow on surfaces of various substrates and penetrate into their inter-particle spaces makes them one of the best adapted species in the use of agro-residues (ViniegraGonzalez and Favela-Torres, 2006) Several lignocellulosic substrates, such as sugarcane bagasse, wheat straw, rice straw, saw dust, corn cobs, corn stover, banana peels, rice husk and orange peel (Iqbal et al.,2011a; Iqbal et al.,2011b; Irshad et al.,2013) have been utilized as potential substrates for βglucosidase production. Cow dung is one of the most abundant and unexploited resource for β-glucosidase production It contains 35.4 % cellulose, 32.6 % hemicelluloses, 13.3 % ash, and 1.4 % nitrogen (Misra et al, 2003). This study was aimed at purifying β-glucosidase produced by Trichoderma viride
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