Abstract

PRODUCTION, PURIFICATION AND CHARACTERIZATION OF AMYLASE USING MICROROGANISM

Rahul A. Pawar*, Anushka Tanavade, Shruti Bhagat, Tanvi Shetye, Siddhi Kudalkar and Vedika Khanvilkar

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Abstract

In this study we investigated the ―Production, Purification, and characterization of Amylase using Microorganism, through the utilization of Starch as a Substrate and Microbial Strain‘ ―Bacillus Subtilis‖. The production of amylase and exposure of the producers to various parameter for the maximum activity of enzyme. Enzyme was produced by the submerged fermentation method. Fermentation experiment conducted at a Controlled temperature range of 35oC to 38oC and orbital shaker speed is 120 rpm. After Fermentation Amylase Activity in the crude enzyme was determined by estimation of the reducing sugar liberated by the action of amylase on soluble starch. Reducing sugar were estimated by 3, 5 Di nitro salicylic acid DNS Method. The production of amylase using microorganism by using factorial design method (23factorial) when factor affecting production like concentration of starch, temperature and ph. were consider. These factors were conducted in 2 levels resulting to 8 Batches the significance of the media was estimated by ANOVA statistics method and the precision of the data was estimated by fit statistics, and it was found that the 48 hours incubation Fermented media 2.5% starch, 38oC temperature and PH-8 showed maximum activity, (Activity: 495.21Mu). Check the maximum activity of crude enzyme after this media as processing to ammonium sulphate precipitation test. After the precipitation of standard DNS assay in media 2.5% starch, PH-8 and temperature 38oC, it was found that at 60% AS saturation showed maximum activity of enzyme. (Activity: 260.7 Mu). this research provides valuable insights into the production of amylase form starch using Bacillus subtilis as the microbial strain. The formulas used to determine amylase activity are also emphasized in this Assay. Every data item in this document is explained with an illustration. This technical study might be essential for the work of the amylase enzyme. The optimization of process parameters like pH, temperature and % of starch and statistical analysis contribute to better understanding of amylase enzyme production and offer potential application in various industries.

Keywords: Amylase, fermentation, Starch, DNS, Bacillus subtilis.