OPTIMIZATION OF CULTURAL PARAMETERS FOR CELLULASE ENZYME PRODUCTION FROM FUNGI

ABSTRACT

The objective of this study is to reduce the production cost of cellulase by optimizing the production medium and using an alternative carbon source such as agricultural solid waste residue. Present studies describe the optimization of process parameters for the production of cellulases by . The fermentation experiments were carried out in shake flasks. Maximum production of cellulases (0.63 U/ml) was observed after a fermentation period of 72 hrs at an temperature of 40° C. Initial pH of the culture medium was also optimized and a pH of 5 was found to support maximum growth and enzyme production (0.9 U/ml) by A. niger. Different inorganic nitrogen sources and carbon sources were evaluated for the production of cellulases and ammonium sulphate in case of nitrogen and glucose among carbon sources was found to be the best. Agrobased coconut cake waste gave the best production of cellulases.

Keywords: Cellulase, Agricultural waste, Fungi, optimization, Aspergillus niger, Penicillium chrysogenum

REFERENCES

1. Kibbelwhite, P.R. and Clark, T.A. 1996. Enzymatic modification of radiata pine kraft fiber and handsheet properties. Appia J.  49:390–396.
2. Jagdish Singh and Pawandeep Kaur. 2012. Optimization of process parameters for cellulase production from Bacillus sp. JS14 in solid substrate fermentation using response surface methodology Braz. arch. biol. technol.  55(4).
3. Beguin, P. and Aubert,  J.P.1994.The biological degration of cellulose. FEMS Microbiol Rev.13: 25-28.
4. Vinod Kumar Nathan, Mary Esther Rani, Gunaseeli Rathinasamy, Kannan Narayanan Dhiraviam and Sridhar Jayavel. 2014.  Process optimization and production kinetics for cellulase production by Trichoderma viride VKF3 SpringerPlus.3:92.
5. Lee, B. H. ,Kim, B. K.., Lee, Y. J., Chung, C. H. and Lee, J. W. 2010. “Industrial scale of optimization for the production of carboxymethylcellulase from rice bran by a marine bacterium, Bacillus subtilis subsp. subtilis A-53,” Enzyme and Microbial Technology, 46(1): 38–42.
6. Ali, U.F. and Saad El-Dein , H.S. 2008. Production and Partial Purification of Cellulase Complex byAspergillus niger and A. nidulans Grown on Water Hyacinth Blend. J Appl Sci Res. 4: 875-891.
7. Chahal, D.S.1985. Solid state fermentation with Trichoderma reesei for cellulose production. Appl Environ Microbiol. 49: 205-210.
8. Koomnok, C. 2005. Selection of cellulose producing thermophilic fungi. 31st congress on Science and Technology of Thailand.
9. Yano, S., Ozaki, H., Matsuo, S., Ito, M., Wakayama, M. and Takagi, K. (2012) Production, purification and characterization of D-aspartate oxidase from the fungus Trichoderma harzianum SKW-36. Adv Biosci Biotechnol 3(1):7–13
10. Penttila, M., Limon, C. and  Nevalainen, H. 2004. Molecular biology of Trichoderma and biotechnological applications, Handbook of fungal biotechnology. Marcel Dekker, New York
11. Kathiresan , K. and Manivannan, S. 2006. Cellulase production by Penicillium fellutanum isolated from coastal mangrove rhizosphere soil. Res J Microbiol. 1(5):438–442
12. Polyanna, N.H., Porto, T.S., Moreira, K.A., Pinto, G.A.S., Cristina, M.S.M. and Ana, L.F.P. 2011. Cellulase production by Aspergillus japonicus URM5620 using waste from castor bean (Ricinus communis L.) under solid state fermentation. Appl Biochem Biotechnol. 165:1057–1067.
13. Hankin, L. and Anagnostakis, S. L. 1977. “Solid media containing carboxymethylcellulose to detect C_xcellulase activity of micro organisms,” Journal of General Microbiology. 98(1):109–115.
14. Sherief, A. A., El-Tanash, A. B., and Atia, N. 2010. “Cellulase production by Aspergillus fumigatus grown on mixed substrate of rice straw and wheat bran,” Research Journal of Microbiology. 5(3):199–211.
15. Miller, G. L. 1959. “Use of dinitrosalicylic acid reagent for determination of reducing sugar,” Analytical Chemistry. 31(3):426–428.
16. Pointing, S. B. 1999. “Qualitative methods for the determination of lignocellulolytic enzyme production by tropical fungi,” Fungal Diversity. 2:17–33.
17. Milala, M.A., Shugaba, A., Gidado, A., Ene, A.C. and Wafar, J.A. 2005. Studies on the use of aricultural wastes for cellulase enzyme production by Aspergillus niger. Research Journal of Agriculture and biological Sciences 4: 323-325.
18. Abo-State, M.A.M., Hammad, A.E., Selin, M. and Gannam, R.B. 2010. Enhanced production of cellulases by Aspergillus sp on agricultural wastes by solid state fermentation. American – Eurasian Journal of Agricultural & Environmental Science. 402 – 410.
19. Beldman, G., Searle-Van, L.M.F., Rombouts, F.M. and Voragen, F.G.J. 1985.The celulase of Trichoderma virde: Purifcation, characterization and comparison of al detectable endoglucanases, exoglucanase and B-glucidase. Eur J Biochem. 146:301-308.
20. Ali, S., Sayed, A., Sarker, R.T., Alam, R. 1991. Factors afecting celulase production by Aspergilus tereus and Aspergilus niger. Enz Microbial Technol. 191(1):606-616.
21. Dopelbauer, R., Esterbauer, H., Steiner, W., Laferty, R. and Steinmuler, H. 1987.The use of celulosic wastes for production of celulases by Trichoderma resei. Appl Microbiol Biotechnol. 26:485-494.
22. Sabu, A., Sarita, S., Pandey, A., Bogar, B., Szakacs, G. and Socol, C.R. Solid-State Fermentation for Production of Phytase by Rhizopus oligosporus. Apl Biochem Biotechnol. 202;103:251-260.
23. Murao, S., Sakamoto, R. and Arai, M. 1988. “Cellulase of Aspergillus aculeatus,” in Methods in Enzymology, W. A. Wood and S. T. Kellog, Eds., vol. 160, pp. 275–284, Academic Press, London, UK.
24. Lu, W., Li, D. and Wu, Y. 2003. “Influence of water activity and temperature on xylanase biosynthesis in pilot-scale solid-state fermentation by Aspergillus sulphureus,” Enzyme and Microbial Technology, 32(2):305–311.
25. Immanuel, G., Akila Bhagavath, C. M. Iyapppa Raj, P., Essakking, P. and Palavessam A. 2007. Production and partial purification of cellulase by Aspergillus niger and A. fumigatus fermented in coir waste and saw dust. Internet Journal for Microbiology 3:1 – 17.
26. Enari, T. M. and Markkanen, P. 1977.  Adv. Biochem. Eng. 5, 1.
27. Acharya, P. B., Acharya, D. K. and  Modi, H. A.  2008. African Journal of Biotechnology. 7 (22) : 4147.
28. Depaula, E.H., Ramos, L. P. and Azevedo, M.D. 1999. The potential  of Humicola grisea var. thermoidea for bioconversion of sugarcane bagasse. Bioresour. Technol.68: 35 – 41.
29. Shin, C.S., Lee, J.P. and Park, S.C. 2000. Enzyme production of Trichoderma reesei Rut C–30 on various lignocellulosic substrates. Appl. Biochem. Biotechnol. 86: 237–245.
30. Szengyel, Z., Zacchi, G., Varga, A. and Reczey, K. 2000. Cellulase production of Trichoderma reesei RUT C30 using steam pretreated spruce. Hydrolytic potential of cellulases on different substrate. Appl. Biochem. Biotechnol. 86: 679–691