Production and Partial Purification of and#945; Amylase From Bacillus cereus By Using Wheat Bran As A Substrate

Abstract

The ubiquitous starch splitting enzyme amylase EC 3.2.1.1 has been a centre of research since decades. It splits starch liberating simple sugars like glucose, maltose. The large scale production of amylase has been carried out using submerged and solid state fermentation Wealth of information exists in support of wheat bran as the best sources among all the agro sources for extracellular amylase production for its higher starch content. newline In the present study Bacillus cereus was isolated from wheat bran and named as WB7. The WB7 isolate has been confirmed as Bacillus cereus by 16srRNA sequencing and phylogenetic trees was created using NJ method Mega 6.0 software. Alpha amylase production from isolated Bacillus cereus was done by solid state fermentation using wheat bran as substrate in aerobic and anaerobic conditions. The aerobic fermentation of Bacillus cereus gave the maximum enzyme activity of alpha amylase 158.6 unit per gram and least by anaerobic fermentation 80.514 unit per gram. The best alpha amylase production activity by Bacillus cereus using wheat bran as substrate shown in Bushnell Hass mineral broth at 37 degree Celsius for 48 hours 101.9 unit per milliliter and least at Nutrient broth at 37 degree Celsius for 48 hours 14.4 unit per milliliter. Hence the Bushnell Hass broth used as selective medium for the production of alpha amylase. The crude enzyme of Bacillus cereus isolate showed total protein 211.18 milligram, specific activity 2199 unit per milligram and yield was 100 percentage. The purified enzyme showed total protein 120.24 milligram, specific activity 799 unit per milligram and yield 20.70 percentage. The observations demonstrated Bacillus cereus as an efficient producer of alpha amylase, that revealed maximum production at temperature 37 degree Celsius.82.1 unit per milliliter, optimum pH 94.1 unit per milliliter, optimum incubation time 48 hours 79.7 unit per milliliter and optimum carbon source glucose 16.1 unit per milliliter. newline newline