Production and optimization of amylase enzyme using bacillus subtilis under solid state fermentation and its characterization

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Year : 2026 | Volume : 04 | 01 | Page :
    By

    Sworna kumari C,

  • R. Kokila,

  • Pavithra Kumar,

  1. Research scholar, Department of biotechnology, University of Madras, Guindy campus, Chennai
  2. Assistant professor, M.M.E.S women’s arts and science college, Melvisharam, Ranipet district, Tamil Nadu, India
  3. Research Scholar, University of Madras, Tamil Nadu, India

Abstract

This study employed solid state fermentation to maximize amylase production by Bacillus subtilis (OR578403). The research began by discovering and analyzing amylase-producing bacteria in soil samples followed with biochemical, molecular characterization. By using commercially available substrates like wheat bran, rice bran, and wheat rava were obtained from the local market and employed as solid substrates, which may influence amylase production and to be assessed. The identified organism was Bacillus subtilis, confirmed through BLAST analysis and phylogenetic tree construction. Further the Bacillus subtilis sample were used in this study to produce amylase, which was then optimized in the fermentation media. Changes in fermentation conditions such as fermentation hour, temperature, inoculum size, carbon sources, metal salts, effect of moisture content and all have a significant effect on enhancing amylase output. Thermal, pH, and effects of metal stability of partly purified amylase were studied and characterized using the SDS PAGE. Finally, From the present study it was considered that this Bacillus subtilis was a potential bacterium producing maximum amylase enzyme.

Keywords: Bacillus Subtilis, Isolation, Phylogenetic Tree, Amylase enzyme, enhanced production with Effect of pH, Temperature, Metal salts, SDS-PAGE characterization

How to cite this article:
Sworna kumari C, R. Kokila, Pavithra Kumar. Production and optimization of amylase enzyme using bacillus subtilis under solid state fermentation and its characterization. International Journal of Biochemistry and Biomolecule Research. 2026; 04(01):-.
How to cite this URL:
Sworna kumari C, R. Kokila, Pavithra Kumar. Production and optimization of amylase enzyme using bacillus subtilis under solid state fermentation and its characterization. International Journal of Biochemistry and Biomolecule Research. 2026; 04(01):-. Available from: https://journals.stmjournals.com/ijbbr/article=2026/view=240827


References

1. Sodhi, H. K., Sharma, K., Gupta, J. K., & Soni, S. K. (2005). Production of a thermostable α-amylase from Bacillus sp. PS-7 by solid state fermentation and its synergistic use in the hydrolysis of malt starch for alcohol production. Process Biochemistry, 40(2), 525-534.

2. Maity, S., Mallik, S., Basuthakur, R., & Gupta, S. (2015). Optimization of solid state fermentation conditions and characterization of thermostable alpha amylase from Bacillus subtilis (ATCC 6633). Journal of Bioprocessing & Biotechniques, 5(4), 1.

3. Hassan, H., & Karim, K. A. (2015). Optimization of alpha amylase production from rice straw using solid-state fermentation of Bacillus subtilis. Int J Sci Environ Technol, 4(1), 1-16.

4. Almanaa, T. N., Vijayaraghavan, P., Alharbi, N. S., Kadaikunnan, S., Khaled, J. M., & Alyahya, S. A. (2020). Solid state fermentation of amylase production from Bacillus subtilis D19 using agro-residues. Journal of King Saud University-Science, 32(2), 1555-1561.

5. Pranay, K., Padmadeo, S. R., & Prasad, B. (2019). Production of amylase from Bacillus subtilis sp. strain KR1 under solid state fermentation on different agrowastes. Biocatalysis and Agricultural Biotechnology, 21, 101300.

6. Swain, M. R., & Ray, R. C. (2007). Alpha‐amylase production by Bacillus subtilis CM3 in solid state fermentation using cassava fibrous residue. Journal of Basic Microbiology, 47(5), 417-425.

7. Akcan, N., Serin, B., & Uyar, F. (2012). Production and optimization parameters of amylases from Bacillus subtilis RSKK96 under solid state fermentation. Chemical and Biochemical Engineering Quarterly, 26(3), 233-239.

8. Mukherjee, A. K., Borah, M., & Rai, S. K. (2009). To study the influence of different components of fermentable substrates on induction of extracellular α-amylase synthesis by Bacillus subtilis DM-03 in solid-state fermentation and exploration of feasibility for inclusion of α-amylase in laundry detergent formulations. Biochemical Engineering Journal, 43(2), 149-156.

9. Paul, M. S., & Sumathy, V. J. H. (2013). Production of amylase from banana peels with Bacillus subtilis using solid state fermentation. Int. J. Curr. Microbiol. App. Sci, 2(10), 195-206.

10. Uyar, F., & Baysal, Z. (2004). Production and optimization of process parameters for alkaline protease production by a newly isolated Bacillus sp. under solid state fermentation. Process Biochemistry, 39(12), 1893-1898.

11. Rameshkumar A, Sivasudha T (2011) Optimization of nutritional constitute for enhanced α-amylase production by solid state fermentation technology. Int J Microbiol Res 2: 143-148.

12. Kandra L (2003) α-Amylases of medical and industrial importance. J Mol Struct (Theochem) 666-667: 487-498.

13. Pandey, P. Nigama, R.S. Carlos, T.S. Vanete, S. Dalel, M. Radjiskumar. Advances in microbial amylases Biotechnolol. Appl. Biochem., 31 (2000), pp. 135-152

14. Suganthi, J.F. Benazir, R. Santhi, V. Ramesh Kumar, A. Hari, N. Meenakashi, K.A. Nidhiya, G. Kavitha, R. Lakshmi.Amylase production by Aspergillus niger under solid state fermentation using agroindustrial wastes.IJEST, 3 (2) (2011), pp. 1756- 1760

15. Valsalam, P. Agastian, M.V. Arasu, N.A. Al-Dhabi, A.K.M. Ghilan, K. Kaviyarasu, B. Ravindran, S.W. Chang, S. Arokiyaraj Rapid biosynthesis and characterization of silver nanoparticles from the leaf extract of Tropaeolum majus L. and its enhanced in- vitro antibacterial, antifungal, antioxidant and anticancer properties, J. Photochem. Photobiol., B: Biol., 191 (2019)

16. Arokiyaraj, M. Saravanan, V. Badathala, Green synthesis of Silver nanoparticles using aqueous extract of Taraxacum officinale and its antimicrobial activity, South Indian J. Biol. Sci., 2 (2015)

17. A. Pandey, C.R. Soccol, D. Mitchell, New developments in solid-state fermentation. I. Bioprocesses and applications, Process. Biochem. 35 (2000) 1153–1169.

18. O. Krik, T.V. Borchert, C.C. Fuglsang, Industrial enzyme applications, Curr. Opin. Biotechnol. 13 (2002) 345–435.

19. R. Gupta, P. Gigras, H. Mohapatra, V.K. Goswami, B. Chauhan, Microbial -amylases: a biotechnological perspectives, Process. Biochem. 38 (2003) 1599–1616.

20. Chandel, R. Rudravaram, V.L. Rao, P. Ravindra, M.L. Narasu, Industrial enzymes in bioindustrial sector development: an Indian perspective, J. Commer. Biotechnol. 3 (2007) 283–291

21. Baysal, F. Uyar, C. Aytekin, Solid state fermentation for production of – amylase by a thermotolerant Bacillus subtilis from hot-spring water, Process. Biochem. 38 (2003) 1663–1668

22. Shukla, R. Kar, Potato peel as a solid state substrate for thermostable -amylase production by thermophilic Bacillus isolates,W. J. Microbiol. Biotechnol. 22 (2006) 417–422

23. Babu, T. Satyanarayana, Alpha-amylase production by thermophilic Bacillus coagulans in solid state fermentation, Process. Biochem. 30 (1995) 305–309.

24. Das, R. Doley, A.K. Mukherjee, Purification and biochemical characterization of a thermostable, alkaliphilic, extracellular -amylase from Bacillus subtilis DM-03, isolated from the traditional fermented food of India, J. Biotechnol. Appl. Biochem. 40 (2004) 291–298.

25. Saha, K., Maity, S., Roy, S., Pahan, K., Pathak, R., Majumdar, S., Gupta, S., 2014. Optimization of amylase production from B. amyloliquefaciens (MTCC 1270) using solid state fermentation. Int. J. Microbiol. 2014, 1–7.

26. Nimkar, M.,D., Deogade, N.,G., Kawale, M., 2010. Production of α-amylase from Bacillus subtilis and Aspergillus niger using different agro-wastes by solid-state fermentation. Asiat. J. Biotechnol. Resour. 01, 23–28.

27. Raul, D., Biswas, T., Mukhopadhyay, S., Kumar Das, S., Gupta, S., 2014. Production and partial purification of alpha amylase from Bacillus subtilis (MTCC 121) using solid state fermentation. Biochem. Res. Int. 2014, 1–5.

28. Tabassum, R., Khaliq, S., Rajoka, M.I., Agblevor, F., 2014. Solid state fermentation of a raw starch digesting alkaline alpha-amylase from Bacillus licheniformis RT7PE1 and its characteristics. Biotechnol. Res. Int. 2014, 1–8.

29. Saxena, R., Singh, R., 2011. Amylase production by solid-state fermentation of agroindustrial wastes using Bacillus sp. Braz. J. Microbiol. 42, 1334–1342. https://dx.doi. org/10.1590/S1517-838220110004000014

30. Singh, R., Shukla, A., Tiwari, S., Srivastava, M., 2014. A review on deligni fi cation of lignocellulosic biomass for enhancement of ethanol production potential. Renew. Sustain. Energy Rev. 32, 713–728. https://doi.org/10.1016/j.rser.2014.01.051.

31. Maity, S., Malik, S., Basuthkur, R., Gupta, S., 2015. Optimization of solid state fermentation conditions ad characterisation of thermostable alpha amylase from Bacillus subtilis ATCC 6633. J. Bioprocess. Biotech. 05. https://doi.org/10.4172 /2155-9821.1000218.

32. Mukherjee, A.K., Borah, M., Rai, S.K., 2009. To study the influence of different components of fermentable substrates on induction of extracellular α-amylase synthesis by Bacillus subtilis DM-03 in solid-state fermentation and exploration of feasibility for inclusion of α-amylase in laundry detergen. Biochem. Eng. J. 43, 149–156. https://doi.org/10.1016/j.bej.2008.09.011.

33. Gangadharan, D., Sivaramakrishnan, S., Nampoothiri, K.,M., Pandey, A., 2006. Solid culturing of Bacillus amyloliquefaciens for alpha amylase production. Food Technol. Biotechnol. 44, 269–274.

34. Sahoo, S., Roy, S., Maiti, S., 2016. A high salt stable ɑ-amylase by Bacillus sp. MRS6 isolated from municipal solid waste; purification, characterization and solid state fermentation. Enzym. Eng. 05, 152. https://doi.org/10.4172/2329-6674.10001502

35. Tanyildizi, M.S., Ozer, D., Elibol, M., 2007. Production of bacterial α-amylase by B. amyloliquefaciens under solid substrate fermentation. Biochem. Eng. J. 37, 294–297. https://doi.org/10.1016/j.bej.2007.05.009.

36. Babu, K.R., Satyanarayana, T., 1995. a-Amylase production by Thermophilic Bacillus coagulans in solid state fermentation. Process Biochem. 30, 305–309. https://doi. org/10.1016/0032-9592(95)87038-5.

37. Sodhi, H.K., Sharma, K., Gupta, J.K., Soni, S.K., 2005. Production of a thermostable α-amylase from Bacillus sp. PS-7 by solid state fermentation and its synergistic use in the hydrolysis of malt starch for alcohol production. Process Biochem. 40, 525–534. In: https://doi.org/10.1016/j.procbio.2003.10.008.

38. Das, K., Doley, R., Mukherjee, A.K., 2004. Purification and biochemical characterization of a thermostable, alkaliphilic, extracellular α-amylase from Bacillus subtilis DM-03, a strain isolated from the traditional fermented food of India. Biotechnol. Appl. Biochem. 40, 291–298. https://doi.org/10.1042/BA20040034.

39. Aiyer, P.V., Modi, H.A., 2005. Isolation and screening of alkaline amylase producing bacterial Bacillus licehniformis SPT-27. Asian Jr. Microbio.Biotech. Env.Sc. 7, 895–897.

Ahead of Print Subscription Review Article
Volume 04
01
Received 12/03/2026
Accepted 31/03/2026
Published 10/04/2026
Publication Time 29 Days
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