• ARUN KUMAR Department of Microbiology, Shoolini University of Biotechnology and Management Sciences, School of Biotechnology, Solan, Himachal Pradesh, India.
  • POONAM KUMARI Department of Microbiology, Shoolini University of Biotechnology and Management Sciences, School of Biotechnology, Solan, Himachal Pradesh, India.
  • KASAHUN GUDETA Department of Applied Biology, Adama Science and Technology University, P.O. Box 1888, Adama, Ethiopia.
  • JM JULKA Department of Biological Sciences, Shoolini University Biotechnology and Management Sciences, School of Biological and Environmental Sciences, Solan, Himachal Pradesh, India.


Objective: The paper aimed to immobilize amylase producing bacterial strain on a suitable matrix and characterization of its physicochemical properties so that much amount of amylase could be produced to be applied in different industries.

Methods: Bacterial colonies were sub-cultured from samples collected from soil in freshly prepared dishes containing starch agar by dot method using sterile inoculating needles from which five different bacteria belonged to genus Bacillus were isolated and assigned as A1, A2, A3, A4, and A5.

Results: It was found that A1 displayed the highest enzyme activity of 17.89 IU/ml with enzyme assay of 0.83 mg/ml and the bacterium was identified to be Bacillus subtilis. A5 displayed 10.13 IU/ml with protein contents of 0.11 mg/ml indicated that A1 possess the highest enzyme activities which were categorized under Bacillus and protein contents and A5 showed less amount of enzyme activities and protein contents as compared to other.

Conclusion: The bacteria which were produced much amount of enzyme activities identified as Bacillus subtilis and recommended and have been recommended to be cultured for the production of amylase enzyme.

Keywords: Amylase, Immobilize, Bacteria, Bacillus, Culture


1. Agrawal M, Pradeep S, Chandraraj K, Gummadi SN. Hydrolysis of starch by amylase from Bacillus sp. KCA102: A statistical approach. Process Biochem 2005;40:2499-507.
2. Pandey A, Soccol CR, Nigam P, Brand D, Mohan R, Roussos S. Biotechnological potential of coffee pulp and coffee husk for bioprocesses. Biochem Eng J 2000;6:153-62.
3. Gupta R, Gigras P, Mohapatra H, Goswami VK, Chauhan B. Microbial ?-amylases: A biotechnological perspective. Process Biochem 2003;38:1599-616.
4. Souza PM. Application of microbial ?-amylase in industry-a review. Braz J Microbiol 2010;41:850-61.
5. Dash S, Clarke G, Berk M, Jacka FN. The gut microbiome and diet in psychiatry: Focus on depression. Curr Opin Psychiatry 2015;28:1-6.
6. Ramakrishnan VA, Thambidurai YU, Rajasekharan SK, Mohanvel SK. Partial characterization and cloning of protease from Bacillus. Asian J Pharm Clin Res 2017;10:187-91.
7. El-Fallal A, Dobara MA, El-Sayed A, Omar N. Starch and microbial ?-amylases: From concepts to biotechnological applications. In: Carbohydrates-Comprehensive Studies on Glycobiology and Glycotechnology; 2012. p. 459-88.
8. Akcan N. High level production of extracellular ?-amylase from Bacillus licheniformis ATCC 12759 in submerged fermentation. Afr J Microbiol Res 2011;5:716.
9. Prescott S, Dunn A. Industrial Microbiology. 4th ed. London: Globe Bookservices; 1983.
10. Abd-Elhalem BT, El-Sawy M, Gamal RF, Abou-Taleb KA. Production of amylases from Bacillus amyloliquefaciens under submerged fermentation using some agro-industrial by-products. Ann Agric Sci 2015;60:193-202.
11. Sivaramakrishna A, Kalikhman I, Kertsnus E, Korlyukov AA, Kost D. Donor-stabilized silyl cations. 10. Pentacoordinate siliconium-ion salts with a triphenylphosphinimino-N ligand group: Two-bond P-N-Si coupling as a measure for coordination strength1. Organometallics 2006;25:3665-9.
12. Rossi MI, Yokota T, Medina KL, Garrett KP, Comp PC, Schipul AH Jr., et al. B lymphopoiesis is active throughout human life, but there are developmental age-related changes. Pediatr Blood Cancer 2003;101:576-84.
13. Konsoula Z, Liakopoulou-Kyriakides M. Alpha-amylase production in aqueous two-phase systems by Bacillus subtilis. FEBS J 2005;272:497.
14. Ashwini K, Gaurav K, Karthik L, Bhaskara Rao KV. Optimization, production and partial purification of extracellular ?-amylase from Bacillus sp. Marini. Arch Appl Sci Res 2011;3:33-42.
15. Alli AI, Ogbonna CI, Rahman AT. Hydrolysis of certain Nigeria: Cereal starches using crude fundal amylase. Niger J Biotechnol 1998;9:24-36.
16. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the folin phenol reagent. J Biol Chem 1951;193:265-75.
17. Pandya PH, Jasra RV, Newalkar BL, Bhatt PN. Studies on the activity and stability of immobilized ?-amylase in ordered mesoporous silicas. Microporous Mesoporous Mater 2005;77:67-77.
18. Kumar RS, Vishwanath KS, Singh SA, Rao AA. Entrapment of ?-amylase in alginate beads: Single step protocol for purification and thermal stabilization. Process Biochem 2006;41:2282-8.
19. Dhanasekaran D. Studies on free and immobilised cells of Bacillus species on the production of alpha-amylase. Int J Microbiol 2006;2:2039-47.
20. Konsoula Z, Liakopoulou-Kyriakides M. Thermostable ?-amylase production by Bacillus subtilis entrapped in calcium alginate gel capsules. Enzyme Microbial Technol 2006;39:690-6.
21. Samanta A, Bera P, Khatun M, Sinha C, Pal P, Lalee A, et al. An investigation on heavy metal tolerance and antibiotic resistance properties of bacterial strain Bacillus sp. Isolated from municipal waste. J Microbiol Biotechnol Res 2012;2:178-89.
22. Yassien MA, Asfour HZ. Improved production, purification and some properties of ?-amylase from Streptomyces clavifer. Afr J Biotechnol 2012;11:14603-11.
23. Mishra S, Behera N. Amylase activity of a starch degrading Bacteria isolated from soil receiving kitchen wastes. Afr J Biotechnol 2008;7:3326-31.
24. Kar N, Roy RN, Sen SK, Ghosh K. Isolation and characterization of extracellular enzyme producing Bacilli in the digestive tracts of rohu, Labeo rohita (Hamilton) and Murrel, channa punctatus (Bloch). Asian Fish Sci 2008;21:421-34.
25. Oyeleke SB, Egwim EC, Auta SH. Screening of Aspergillus flavus and Aspergillus fumigatus strains for extracellular protease enzyme production. J Microbiol Antimicrob 2010;2:83-7.
26. Oboh G. Isolation and characterization of amylase from fermented cassava (Manihot esculenta Crantz) wastewater. Afr J Biotechnol 2005;4:71347.
27. Ahmed SA, El-Sayed MM, Hassan OK, Saleh NA, Goda HA. Studies on the activity and stability of immobilized Bacillus acidocaldarius alpha-amylase. Aust J Basic Appl Sci 2008;2:466-74.
28. Poddar A, Gachhui R, Jana SC. Optimization of physico-chemical condition for improved production of hyperthermostable ? amylase from Bacillus subtilis DJ5. J Biochem Technol 2012;3:370-4.
29. Beshay U. Production of alkaline protease by Teredinobacter turnirae cells immobilized in Ca-alginate beads. Afr J Biotechnol 2003;2:60-5.
30. Kumar S, Karan R, Kapoor S, Singh SP, Khare SK. Screening and isolation of halophilic bacteria producing industrially important enzymes. Braz J Microbiol 2012;43:1595-603.
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How to Cite
KUMAR, A., P. KUMARI, K. GUDETA, and J. JULKA. “IMMOBILIZATION OF AMYLASE PRODUCING BACTERIA (BACILLUS SUBTILIS) FROM SOIL OF WESTERN HIMALAYAN REGION SOLAN, HIMACHAL PRADESH, INDIA”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 14, no. 2, Feb. 2021, pp. 109-15, doi:10.22159/ajpcr.2021.v14i2.40036.
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