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.

Received: March 29, 2021; Published: May 17, 2021

Abstract

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 50^oC and 60^oC 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 K_m value of 7.9 mg/ml and 3.4mg/ml with a V_max 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

References

1. Ahmed BM., et al. “Potential impact of climate change on termite distribution in africa”. Britain Journal of Environmental Climate Change 1 (2011): 172-189.
2. Al-Kharousi MM., et al. “Characterization of cellulase enzyme produced by chaetomium Sp”. Eurasian Journal of Bio-Sciences 9 (2015): 52-60.
3. Dashtban M., et al. “Cellulase activities in biomass conversion: measurement methods and comparison”. Critical Reviews in Biotechnology 30 (2010): 302-309.
4. Karma M and Ray RR. “Current trends in research and application of microbial cellulase”. Journal of Microbiology 6 (2014): 41-53.
5. Sharada R., et al. “Applications of Cellulases: Review”. International Journal of Pharmaceutical, Chemical and Biological Science 4 (2014): 424-437.
6. Tischer W and Wedekind F. “Immobilized enzymes: Methods and applications”. Topics in Current Chemistry 200 (2015): 96-126.
7. Rodrigues RC., et al. “Modifying enzyme activity and selectivity by immobilization”. Chemical Society Review 42 (2013): 6290-6307.
8. Almin KE., et al. “Extracellular enzyme system utilized by the Fungus Sporotirichum pulverulentum (Chrysospriumlignorum) for the breakdown of cellulose- 2 Activites of the five endo-1,4-beta-glucanases towards carboxymethyl cellulose”. European Journal of Biochemistry 51 (1975): 207-211.
9. Agustini L., et al. “Isolation and Characterization of C cellulase and Xylanase Producing Microbes Isolated from Tropical forests in java and Sumatra”. International Journal of Environment and Bioengineering 3 (2012): 154-167.
10. Uedam MM., et al. “A novel cold adopted cellulase complex from Eiseniafoetida; Characterization of a multienzyme complex with Carboxymethyl cellulase, β-glucanase”. Molecular Biology 157 (2010): 26-32.
11. Paul C O., et al. “Partial purification and characterization of cellulases from digestive tracts of the African giant snail (Achatinaachatina)”. Turkish Journal of Biology (2013): 9-17.
12. Bignell D E. “Introduction to symbiosis”. In: Termites: Evolution, Sociality and Ecology. Kluwer Academic Publishers, Dordrecht, the Netherlands (2000): 189-208.
13. Abe T., et al. “Termites: Evolution, Socially, Symbioses, Ecology”. Kluwer Academic Publishers (2000): 256.
14. Korb J and linsenmair KE. “Experimental heating of macrotermes bellicosus mounds; what does microclimate play in influencing mound” 45 (1998): 335-342.
15. Bradford MM. “A rapid and sensitive method for the quantitation of Microgram quantities of protein utilizing the principle of protein-dye binding”. Analytical Biochemistry 72 (1976): 248-254.
16. Zhang YHP., et al. “Outlook of cellulase improvement: screening and selection strategies”. Biotechnological Advance 24 (2006): 452-481.
17. Watanabe H., et al. “Site of Secretion and Properties of Endogenous Endo-/3-1,4-Glucanase Components from Reticulitermessperatus (Kolbe), a Japanese Subterranean Termite”. Insect Biochemistry and Molecular Biology 4 (2010): 305-313.
18. Fagbohunka B S., et al. “Activities of a Cellulase of the Termite, Ametermes eveuncifer (Silverstri) Soldier: Clue to Termites Salt activity and selectivity by immobilization”. Chemical Society Review 42 (2015): 6290-6307.
19. Fujita A., et al. “Differences in cellulose digestive systems among castes in two termite lineages”. Physiological Entomology 33.1 (2008): 73-82.
20. Fagbohunka B S., et al. “Activities of a Cellulase of the Termite, Ametermes eveuncifer (Silverstri) Soldier: Clue to Termites Salt Intolerance”. Journal of Natural Sciences Research 11 (2016): 117-123.
21. Muhammad NR., et al. “An over view of technologies for immobilization of enzymes and surface analysis techniques forimmobilization of enzymes”. Biotechnology and Biotechnological Equipment2 (2015): 1-6.
22. Hulya T., et al. “Comparison of some properties of free and immobilized cellulase from Baccillus subtilisin calcium alginate beads”. Journal of Biotechnology 38 (2008): 13-23.
23. Viet TO., et al. “Immobilization of cellulase enzyme in calcium alginate gel and its immobilized stability”. American Journal of Research Communication12 (2013): 254-267.
24. Simon P., et al. “Cellulase immobilization on poly (methyl methacrylate) nanoparticles by mini emulsion polymerization”. Brazilian Journal of Chemical Engineering2 (2018): 649-658.
25. Su E., et al. “Immobilization of b-glucosidase and its aroma-increasing effect on tea beverage”. Food Bioproduction Process 88 (2010): 83-89.
26. Rahnama N., et al. “Production and characterisation of cellulase from solid state fermentation of rice straw by Trichodermaharzianum SNRS3”. Pertanika Journal of Tropical Agricultural Science4 (2016): 507-531.
27. Zhou YT., et al. “Improving the Stability of Cellulase by Immobilization on Chitosan-Coated Magnetic Nanoparticles Modified a-Ketoglutaric Acid”. Bioinformatics and Biomedical Engineering1 (2009): 10-14.
28. Kennedy JF., et al. “Transition metal methods for immobilization of enzymes and cells”. Immobilization of Enzymes and Cells. Springer, New York, USA (2012): 345-359.

Citation

Citation: Alhassan AJ., et al. “Effects of Immobilization on the Kinetic Parameters of Partially Purified Cellulase from Termites (Macrotermes bellicosus)".Acta Scientific Pharmacology 2.6 (2021): 20-28.

Copyright

Copyright: © 2020 Alhassan AJ., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.