PRODUCTION AND OPTIMIZATION OF Α-AMYLASE FROM ASPERGILLUS FLAVUS UNDER SOLID STATE FERMENTATION
Abstract
Alpha amylase is one of the important and well known industrial enzymes that cause the breakdown of starch or glycogen. Use of microorganism for the production of amylase is economical as microbes are easy to manipulate to obtain enzymes of desired characteristics. However fungus is preferred over bacteria for enzymes production because of its filamentous nature. Agricultural wastes byproducts have great potential for the production of enzymes. Two indigenous carbon sources namely wheat bran and rice bran were used as substrate for amylase synthesis by Aspergillus flavus through solid state fermentation. Wheat bran gave the highest (670 ± 36μmol./min/mL) and Rice bran (428 ± 24μmol./min/mL) gave the lowest enzyme activity was obtained. Optimum inoculum size of 10% (660 ± 36μmol./min/mL), incubation period 120 hours (670 ± 28 μmol./min/mL), pH 6 (680 ± 42μmol./min/mL), temperature at 30°C (660 ± 46μmol./min/mL) gave highest yield of enzyme production. The highest yield of amylase production was obtained by the addition of MgSO4 0.2% and CaCl2 0.02% respectively. The results obtained in the present study suggest that the Aspergillus flavus may act as a potent strain for industrial production of α-amylase.
Keywords:
Aspergillus flavus, α-amylase, Solid state fermentation, Optimization of parametersDOI
https://doi.org/10.25004/IJPSDR.2015.070313References
2. Anto H, Ujjval T, Kamlesh P. Alpha amylase production by Bacillus cereus MTCC 1305 Using solid state fermentation. Food Technol Biotechnol. 2006; 44:241-245.
3. Pandey A, webb C , Soccol CR, Larroche C. Rice bran as a substrate for proteolytic enzyme production. Enzyme technology. New delhi, Asiatech publishers, 2005.
4. Monga M, Goyal M, Kalra KL, Soni G. Production and stabilization of amylases from Aspergillus niger. Mycosphere. 2011; 2:129-34.
5. Burhan A, Nisa U, Gokhan C, Omer C, Ashabil A, Osman G. Enzymatic properties of a novel thermostable, thermophilic, alkaline and chelator resistant amylase from an alkaliphilic Bacillus sp. isolate ANT-6, Process. Biochem. 2003; 38:1397-1403.
6. Abu EA, Ado SA, James DB. Raw starch degrading amylase production by mixed culture of Aspergillus niger and Saccharomyces cerevisae grown on Sorghum pomace. Afr J Biotechnol. 2005; 4:785-790.
7. Suganthi R, Benazir JF, Santhi R, Ramesh kumar V, Anjana H, Nitya M, Nidhiya KA, Kavitha G, Lakshmi R. Amylase production by aspergillus niger under solid state fermentation using agro industrial wastes. Int J Eng Sci Tech. 2011; 3:1756-1763.
8. Ramachandran S, Patel AK, Nampoothiri KM, Chandran S, Szakacs G, Soccol CR, Pandey A. Alpha amylase from a fungal culture grown on oil cakes and its properties. Brazilian Archives of Biology and Technology. 2004; 47:309-317.
9. Lonsane BK, Ghildyal NP, Budiatman S, Ramakrishna SV. Engineering aspects of solid state fermentation. Enzyme Microb Technol. 1985; 7:258-265.
10. Baysal, Z, Uyar F, Aytekin C. Solid-state fermentation for production of α-amylase by a thermotolerant Bacillus subtilis from hot-spring water. Process Biochem. 2003; 38:1665-1668.
11. Arasaratnam V, Mylvaganam K, Balasubramaniam K. Glucoamylase production by Aspergillus niger in solid state fermentation with paddy husk as support. J Food Sci Technol. 2001; 38:334-338.
12. Kocher GS, Kaur P, Grewal HS. Production of α-amylase by Aspergillus niger using wheat bran in submerged and solid state fermentations. Indian J Microbiol. 2003; 43:143–145.
13. Tanyildizi MS, Ozer D, Elibol M. Optimization of a-amylase production by Bacillus sp. using response surface methodology. Process Biochem. 2005; 40:2291-2296.
14. Carlsen M, Nielsen J, Villadsen J. Growth and a-amylase production by Aspergillus oryzae during continuous cultivations. J. Biotechnol. 1996; 45:81-93.
15. Asgher M, Asad MJ, Rahman SU, Legge RL. A thermostable - amylase from a moderately thermophilic Bacillus subtilis strain for starch processing. J Food Eng. 2007; 79:950-955.
16. Sadasivam S, Manickam A. Estimation of starch by antrone reagent. Biochemical Methods: New Age International Publishers, 1996.
17. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin-Phenol reagents. Journal of Biological Chemistry. 1951; 48:17-25.
18. Alva S, Anupama J, Salva J, Chiu YY, Vyshali P, Shruthi M, Yogeetha BS, Bhavya D, Purvi J, Ruchi K, Kumudini BS, Varalakshmi KN. Production and characterization of fungal amylase enzyme isolated from Aspergillus sp. JGI 12 in solid state culture. African journal of Biotechnology. 2007; 6:576-581.
19. Miller GL. Use of dinitro-salicyclic acid for the determination of reducing sugars. Analytical chemistry. 1959; 31:426.
20. Sharma DK, Tiwari M, Behere BK. Solid state fermentation of new substrates for production of cellulase and other biopolymer hydrolyzing enzymes. Appl Biochem Biotechnol. 1996; 15:495-500.
21. Ammar MS and El-Safey EM. Production of α-amylase enzyme produced by Aspergillus flavus var. columnaris, S-9KP from maize meal and rice husk under solid-state fermentation (SSF) conditions in open air. International Conference of Enzymes in the Environment, Activity, Ecology and Applications, Praha, Czech Republic, 2003, pp.33.
22. Zambare V. Solid state fermentation of Aspergillus oryzae for Glucoamylase Production on Agro residues. Int J Life Sci. 2010; 4:16-25.
23. Silva S, Elmore BB, Houston KH. Cellular activity of Trichoderma ressei (RUT-C30) on municipal solid waste. Applied Biochemistry and Biotechnology 1995; 15:145-153.
24. Kathiresan K, Manivannan S. α--amylase production by Penicillium fellutanum isolated from mangrove rhizosphere soil. African Journal of Biotechnology 2006; 5:829-832.
25. Kundu AK, Das S, Gupta TK. Influence of culture and nutritional conditions on the production of amylase by the submerged culture of Aspergillus oryzae. Journal of Fermentation Technology 1973; 51:142-150.
26. Castro PML, Hayter PM, Ison AP, Bull AT. 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 1992; 38:84-90.
27. Gupta R, Gigras P, Mohapatra H, Goswami V K, Chauhan B. Microbial amylases: a biotechnological perspective. Process Biochemistry 2003; 38:1599-1616.
28. Francis FA, Sabu KM, Nampoothiri S, Ramachandran S, Ghosh S, Szakacs G, Pandey A. Use of response surface methodology for optimizing process parameters for the production of a amylase by Aspergillus oryzae. Biochem Eng J. 2003; 15:107-115.
29. Gupta RP, Gigras H, Mohapatra VK, Goswami B, Chauhan B. Microbial α -amylases: a biotechnological perspective. Process Biochem. 2003; 38:1599-1616.
30. Souza DFD, Peralta RM. Production of amylases by Aspergillus tamarii in solid state fermentation at high initial glucose concentrations. Acta Scientiarum Maringa. 2001; 23:599-602.
31. Sindhu R, Suprabha GN, Shashidhar S. Optimization of process parameters for the production of a- amylase from Penicillium janthinellum (NCIM 4960) under solid state fermentation. African Journal of Microbiology Research 2009; 3:498-503.
Published

