Production and Purification of Amylase from Bacillus subtilis Isolated from Soil

  • G. Kalyani
  • E.M. Rajesh
Keywords: Bacillus Subtilis, Amylase, Ammonium Sulphate Fractionation, Dialysis, DEAE-Cellulose Column Chromatography


In spite of progress in biotechnology and enzymology, the enzymes have been industrialized in recent years for the mounting up the product development in various arena. The ultimate goal of this study comprises the production and purification the amylase enzyme from the bacterial strain. A powerful amylase producer, Bacillus subtilis ISOLATE-4 was isolated, screened and identified from the soil sample. In order to produce extracellular amylase, various physico-chemical parameters were optimized. During optimization, the maximal production of amylase by the isolate at 48 hrs of incubation in 100 rpm was found to be 6.93U/ml, 5.94U/ml, 6.0U/ml at 45ºC, pH 6 with 1% substrate concentration respectively. Ammonium sulphate fractionation was done for rapid precipitation of the amylase at a concentration of 60% and exposed to dialysis showed the 25% purification fold of an enzyme. The dialyzed product was further subjected to DEAE-Cellulose column chromatography resulted in an increase upto 75% purification fold than crude enzyme. The amylase enzyme might be suitable for the liquefaction of starch, detergent, textile and several additional industrial applications.


Download data is not yet available.


Castro GR, Ferrero MA, Mendez BS, & Sineriz F. (1993). Screening and selection of bacteria with high amylolytic activity. Acta Biotechnologica Banner, 13(2), 197-201.

Chakraborty, S., Khopade, A., Kokare, C., Mahadik, K., & Chopade, B. (2009). Isolation and characterization BACTERIA of novel α-amylase from marine Streptomyces sp. D1. Journal of Molecular Catalysis B: Enzymatic, 58(1), 17-23.

Chand, N., Sajedi, R. H., Nateri, A. S., Khajeh, K., & Rassa, M. (2014). Fermentative desizing of cotton fabric using an α-amylase-producing Bacillus strain: Optimization of simultaneous enzyme production and desizing. Process Biochemistry, 49(11), 1884-1888.

Chung, Y.C., Kobayashi, T., Kanai, H., Akiba, T., & Kudo, T. (1995). Purification and properties of extracellular amylase from the hyperthermophilic archaeon thermococcus profundus DT 5432. Applied Environmental Microbiology, 61(4), 1502-1506.

Dhiman, S. S., Sharma, J., & Battan, B. (2008). Industrial applications and future prospects of microbial xylanases: A review. Bio Resources, 3(4), 1377-1402.

Gangadharan D, Sivaramakrishnan S, Namboothiri KM, & Pandey A. (2006). Solid culturing of Bacillus amyloliquefaciens for -amylase production. Food Technology Biotechnology, 44(2), 269-274.

Gupta R, Gigras P, Mohapatra H, Goswami VK, & Chauhan B (2003). Microbial α-amylases: A biotechnological prospective. Process Biochemistry, 38(11), 1599-1616.

How to Cite
G. Kalyani, & E.M. Rajesh. (2018). Production and Purification of Amylase from Bacillus subtilis Isolated from Soil. International Journal of Engineering and Management Research, 8(3), 246-254.