Isolation, screening of Aspergillus flavus and its production parameters for á- amylase under solid state fermentation
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Abstract
The amylase producing fungi were isolated from spoiled fruits, vegetables and soil, in and around Bangalore, Karnataka, India. The isolates were identified and five fungal species were screened. The best amylase producer among them, Aspergillus sp was selected for enzyme production by both sub merged fermentation using mineral salt medium (MSM) and solid state fermentations using wheat bran as a solid substrate. The various parameters influencing solid state fermentation were optimized. The most important factors are such as pH, incubation temperature, incubation period, carbon sources, nitrogen sources and moisture content. The maximum amount of enzyme production was obtained when solid state fermentation was carried out with soluble starch as carbon source and beef extract (1% each) as nitrogen source, optimum conditions of pH 7.0, an incubation temperature of 25 (±2) °C, incubation time 96 h and 62% moisture content.
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Keywords
Isolation, Screening, Production, á -amylase, Aspergillus flavus, Solid state fermentation
References
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Alva, S., Anupama, J., Salva, J., Chiu, Y.Y., Vyshali, P., Shruthi, M., Yogeetha, B.S., Bhavya, D., Purvi, J., Ruchi, K., Kumudini, B.S., and Varalakshmi, K. N. (2007). Production and characterization of fungal amylase enzyme isolated from Aspergillus sp. JGI 12 in solid state culture. African journal of Biotechnology, 6: 576 – 581.
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Baysal, Z., Uyar, F. and Aytekin, C. (2003). Solid-state fermentation for production of amylase by a thermotolerant Bacillus subtilis from hot-spring water. Process Biochemistry, 38: 1665–1668.
Bhat, M.K. (2000). Cellulases and related enzymes in biotechnology. Biotechnology Advances, 18: 355-383.
Bhatti, H.N., Rashid, M.H., Nawaz, R., Asgher, M., Perveen, R., and Jabbar, A. (2007). Optimization of Media for Enhanced Glucoamylase Production in Solid-State Fermentation by Fusarium solani. Food Technology and Biotechnology, 45 (1): 51–56.
Cappuccino, J.G. and Sherman, N. (2004). Microbiology a laboratory manual, (6th edition) Pearson Education, Pvt. Ltd, Delhi, India.
Castro, P. M. L., Hayter, P. M., Ison, A. P. and Bull, A.T. (1992). Application of statistical design to the optimization of culture medium for recombinant interferon-gamma Process Biochemistry production by Chinese hamster ovary cells. Applied Microbiology and Biotechnology, 38: 84-90.
Chimata, M. K., Sasidhar, P. and Challa, S. (2010). Production of extracellular amylase from agricultural residues by a newly isolated Aspergillus species in solid state fermentation. African Journal of Biotechnology, 9 (32): 5162-5169.
Ellaiah, P., Adinarayana, K., Bhavani, Y., Padmaja, P. and Srinivasulu, B. (2002). Optimization of process parameters for glucoamylase production under solid state fermentation by a newly isolated Aspergilus species. Process Biochemistry, 38: 615–620.
Erdal, S. and Taskin, M. (2010). Production of á-amylase by Penicillium expansum MT-1 in solid-state fermentation using waste Loquat (Eriobotrya japonica Lindley) kernels as substrate. Romanian Biotechnological Letters, 15 (3):5342-5350.
Gupta, R., Gigras, P., Mohapatra, H., Goswami, V. K. and Chauhan, B. (2003). Microbial -amylases: a biotechnological perspective. Process Biochemistry, 38: 1599-1616.
Kathiresan, K., and Manivannan, S. (2006). a-amylase production by Penicillium fellutanum isolated from mangrove rhizosphere soil. African Journal of Biotechnology, 5 (10): 829-832.
Kundu, A.K., Das, S. and Gupta, T. K. (1973). Influence of culture and nutritional conditions on the production of amylase by the submerged culture of Aspergillus oryzae. Journal of Fermentation Technology, 51: 142-150.
Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R. J., (1951). Protein measurement with the Folin-Phenol reagents. Journal of Biological Chemistry, 48: 17-25.
Michelena, V.V. and Castillo, F.J. (1984). Production of amylase by Aspergillus foetidus on rice flour medium and characterization of the enzyme. Journal of Applied Bacteriology, 56 : 395–407.
Miller,G. L. (1959). Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar. Analytical Chemistry, 31 (3): 426–428.
Pandey, A., Soccol, C.R., Nigam, P., Brand, D., Mohan, R. and Roussos, S. (2000). Biotechnological potential of coffee pulp and coffee husk for bioprocesses. Biochemical Engineering Journal, 6: 153–162.
Patel, A. K., Nampoothiri, K. M., Ramachandran, S., Szakacs, G. and Pandey, A. (2005). Partial purification and characterization of a-amylase produced by Aspergillus oryzae using spent brewing grains. Indian Journal of Biotechnology, 4:336–341.
Ramachandran, S., Patel, A. K., Nampoothiri, K. M., Francis, F., Nagy, V., Szakacs, G. and Pandey, A. (2004). Coconut oil cake––a potential raw material for the production of aamylase. Bioresource Technology, 93: 169–174.
Silva, S., Elmore, B. B. and Houston K. H. (1995). Cellular activity of Trichoderma ressei (RUT-C30) on municipal solid waste. Applied Biochemistry and Biotechnology, 15: 145-153.
Sindhu, R., Suprabha, G. N., and Shashidhar, S. (2009). Optimization of process parameters for the production of a- amylase from Penicillium janthinellum (NCIM 4960) under solid state fermentation. African Journal of Microbiology Research, 3 (9): 498-503.
Souza, D.F.D. and Peralta, R.M. (2001). Production of amylases by Aspergillus tamarii in solid state fermentation at high initial glucose concentrations. Acta Scientiarum Maringa, 23, (2): 599-602.
Sun, X., Liu, Z., Qu, Y. and Li, X. (2007).The effects of wheat bran composition on the production of biomass-hydrolyzing enzymes by Penicillium decumbens. Applied Biochemistry and Biotechnology, 146: 119-128.
Varalakshmi, K. N., Kumudini, B.S., Nandini, B.N., Solomon, J., Suhas, R., Mahesh, B and Kavitha, A.P. (2009). Production and characterization of a-amylase from Aspergillus niger JGI 24 isolated in Bangalore. Polish Journal of Microbiology, 58: 29-36.
Vries, R.P.D. and Visser, J. (2001). Aspergillus enzymes involved in degradation of plant cell wall polysaccharides. Microbiology and Molecular Biology reviews, 65 (4): 497–522.
Whiteley,C.G., and Lee, D.J., (2006). Enzyme technology and biological remediation. Enzyme and Microbial Technology, 38: 291-316.
Yabuki, M., Ono, N., Hoshino, K. and Fukui. S., (1977). Rapid induction of amylase by non-growing mycelia of Aspergillus oryzae. Applied and Environmental Microbiology, 34: 1-6.
Alva, S., Anupama, J., Salva, J., Chiu, Y.Y., Vyshali, P., Shruthi, M., Yogeetha, B.S., Bhavya, D., Purvi, J., Ruchi, K., Kumudini, B.S., and Varalakshmi, K. N. (2007). Production and characterization of fungal amylase enzyme isolated from Aspergillus sp. JGI 12 in solid state culture. African journal of Biotechnology, 6: 576 – 581.
Aneja, K.R. (2010). Experiments in Microbiology, Plant Pathology and Biotechnology, New age International publishers, New Delhi.
Baysal, Z., Uyar, F. and Aytekin, C. (2003). Solid-state fermentation for production of amylase by a thermotolerant Bacillus subtilis from hot-spring water. Process Biochemistry, 38: 1665–1668.
Bhat, M.K. (2000). Cellulases and related enzymes in biotechnology. Biotechnology Advances, 18: 355-383.
Bhatti, H.N., Rashid, M.H., Nawaz, R., Asgher, M., Perveen, R., and Jabbar, A. (2007). Optimization of Media for Enhanced Glucoamylase Production in Solid-State Fermentation by Fusarium solani. Food Technology and Biotechnology, 45 (1): 51–56.
Cappuccino, J.G. and Sherman, N. (2004). Microbiology a laboratory manual, (6th edition) Pearson Education, Pvt. Ltd, Delhi, India.
Castro, P. M. L., Hayter, P. M., Ison, A. P. and Bull, A.T. (1992). Application of statistical design to the optimization of culture medium for recombinant interferon-gamma Process Biochemistry production by Chinese hamster ovary cells. Applied Microbiology and Biotechnology, 38: 84-90.
Chimata, M. K., Sasidhar, P. and Challa, S. (2010). Production of extracellular amylase from agricultural residues by a newly isolated Aspergillus species in solid state fermentation. African Journal of Biotechnology, 9 (32): 5162-5169.
Ellaiah, P., Adinarayana, K., Bhavani, Y., Padmaja, P. and Srinivasulu, B. (2002). Optimization of process parameters for glucoamylase production under solid state fermentation by a newly isolated Aspergilus species. Process Biochemistry, 38: 615–620.
Erdal, S. and Taskin, M. (2010). Production of á-amylase by Penicillium expansum MT-1 in solid-state fermentation using waste Loquat (Eriobotrya japonica Lindley) kernels as substrate. Romanian Biotechnological Letters, 15 (3):5342-5350.
Gupta, R., Gigras, P., Mohapatra, H., Goswami, V. K. and Chauhan, B. (2003). Microbial -amylases: a biotechnological perspective. Process Biochemistry, 38: 1599-1616.
Kathiresan, K., and Manivannan, S. (2006). a-amylase production by Penicillium fellutanum isolated from mangrove rhizosphere soil. African Journal of Biotechnology, 5 (10): 829-832.
Kundu, A.K., Das, S. and Gupta, T. K. (1973). Influence of culture and nutritional conditions on the production of amylase by the submerged culture of Aspergillus oryzae. Journal of Fermentation Technology, 51: 142-150.
Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R. J., (1951). Protein measurement with the Folin-Phenol reagents. Journal of Biological Chemistry, 48: 17-25.
Michelena, V.V. and Castillo, F.J. (1984). Production of amylase by Aspergillus foetidus on rice flour medium and characterization of the enzyme. Journal of Applied Bacteriology, 56 : 395–407.
Miller,G. L. (1959). Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar. Analytical Chemistry, 31 (3): 426–428.
Pandey, A., Soccol, C.R., Nigam, P., Brand, D., Mohan, R. and Roussos, S. (2000). Biotechnological potential of coffee pulp and coffee husk for bioprocesses. Biochemical Engineering Journal, 6: 153–162.
Patel, A. K., Nampoothiri, K. M., Ramachandran, S., Szakacs, G. and Pandey, A. (2005). Partial purification and characterization of a-amylase produced by Aspergillus oryzae using spent brewing grains. Indian Journal of Biotechnology, 4:336–341.
Ramachandran, S., Patel, A. K., Nampoothiri, K. M., Francis, F., Nagy, V., Szakacs, G. and Pandey, A. (2004). Coconut oil cake––a potential raw material for the production of aamylase. Bioresource Technology, 93: 169–174.
Silva, S., Elmore, B. B. and Houston K. H. (1995). Cellular activity of Trichoderma ressei (RUT-C30) on municipal solid waste. Applied Biochemistry and Biotechnology, 15: 145-153.
Sindhu, R., Suprabha, G. N., and Shashidhar, S. (2009). Optimization of process parameters for the production of a- amylase from Penicillium janthinellum (NCIM 4960) under solid state fermentation. African Journal of Microbiology Research, 3 (9): 498-503.
Souza, D.F.D. and Peralta, R.M. (2001). Production of amylases by Aspergillus tamarii in solid state fermentation at high initial glucose concentrations. Acta Scientiarum Maringa, 23, (2): 599-602.
Sun, X., Liu, Z., Qu, Y. and Li, X. (2007).The effects of wheat bran composition on the production of biomass-hydrolyzing enzymes by Penicillium decumbens. Applied Biochemistry and Biotechnology, 146: 119-128.
Varalakshmi, K. N., Kumudini, B.S., Nandini, B.N., Solomon, J., Suhas, R., Mahesh, B and Kavitha, A.P. (2009). Production and characterization of a-amylase from Aspergillus niger JGI 24 isolated in Bangalore. Polish Journal of Microbiology, 58: 29-36.
Vries, R.P.D. and Visser, J. (2001). Aspergillus enzymes involved in degradation of plant cell wall polysaccharides. Microbiology and Molecular Biology reviews, 65 (4): 497–522.
Whiteley,C.G., and Lee, D.J., (2006). Enzyme technology and biological remediation. Enzyme and Microbial Technology, 38: 291-316.
Yabuki, M., Ono, N., Hoshino, K. and Fukui. S., (1977). Rapid induction of amylase by non-growing mycelia of Aspergillus oryzae. Applied and Environmental Microbiology, 34: 1-6.
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How to Cite
Isolation, screening of Aspergillus flavus and its production parameters for á- amylase under solid state fermentation. (2011). Journal of Applied and Natural Science, 3(2), 268-273. https://doi.org/10.31018/jans.v3i2.194
