Effects of Immobilization on the Kinetic Parameters of Partially Purified Cellulase from Termites (Macrotermes bellicosus)
Gadaka MA1, Alhassan AJ2*, Zainab GA2, Muhammad YY1, Muhammad IU3, Muazu AB3 and Murtala M2
1Department of Biochemistry, Faculty of Science, University of Maiduguri, Nigeria
2Department of Biochemistry, Faculty of Basic Medical Sciences Bayero University, Kano, Nigeria
3Department of Biochemistry, College of Medical Sciences, Yobe State University, Damaturu, Yobe State, Nigeria
*Corresponding Author: Alhassan AJ, Department of Biochemistry, Faculty of Basic Medical Sciences Bayero University, Kano, Nigeria.
March 29, 2021; Published: May 17, 2021
Cellulase is a class of hydrolase enzymes with commercial and industrial value, it is commonly produced by the microorganism such as fungi, protozoans, bacteria and even insect that survives on cellulose. In this study, we evaluated the effect of immobilization on the kinetic parameters of partially purified cellulase from Macrotermes bellicosus. The cellulase was extracted from matured M. bellicosus and subjected the supernatant of the crude extract that has cellulase activity to gel filtration and ion-exchange chromatography. The enzyme was partially purified 2.0 fold with an overall yield of 40.4% on DEAE- cellulose column and a final specific activity of 51.0U/mg. The partially purified cellulase was immobilized by entrapment on calcium alginate beads. The free and immobilized enzyme showed an optimum temperature of 50oC and 60oC and optimum pH of 6.0 and 8.0 respectively. Initial velocity studies for the determination of kinetic constants with cellulose as a substrate revealed a Km value of 7.9 mg/ml and 3.4mg/ml with a Vmax value of 1.59 unit/mg and 1.15 unit/mg for the free and immobilized enzyme respectively. Both the free and immobilized cellulase activity was enhanced by Ca2+, and Mn2+ but slightly decreased by Na+. While Mg2+ and Zn2+ were found to be strong inhibitors of both the free and immobilized enzyme. This research shows that cellulase from M. bellicosus could be immobilized and utilized for degradation of cellulose-containing materials because of their high catalytic activity, thermostability and acid-base stability, which reflect the potential industrial significance of the enzyme.
Keywords: Cellulase; Macrotermes bellicosus; Partial Purification; Immobilization
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