CYCLODEXTRIN GLYCOSYLTRANSFERASE PRODUCTION BY ALKALIPHILIC BACILLUS SP. ISOLATED FROM RICE CULTIVATED SOIL AND MEDIA OPTIMIZATION USING TAGUCHI METHODAbstract
A Cyclodextrin glycosyltransferase (CGTase) producer was isolated from soil obtained from rice fields using alkaline Horikoshi II medium. The alkaliphile was characterized microscopically, biochemically and confirmed by 16S rRNA analysis as Bacillus oshimensis. CGTase production is dependent on the strain, medium composition and culture conditions; hence media optimization studies were carried out in shake flask using the statistical method. Tapioca starch was used as carbon source and a combination of yeast extract and peptone were used as nitrogen source in the media, along with MgSO4.7H2O, K2HPO4 and Na2CO3. A 33 factorial design using Taguchi method has been chosen to elucidate the combined effect of these variables. The parameters were optimized by changing three independent variables, with fixed concentration of mineral sources. The optimal media composition for CGTase production was found to be comprised of Tapioca starch 1%; peptone 0.5%; yeast extract 0.5% and 0.14% Magnesium Sulphate, 0.2% ammonium dihydrogen phosphate according to the factorial designing of the experiment and was used for further studies. The CGTase activity was found to be 4.5 U/ml after media optimization.
R. Kamble and A. Gupte*
Department of Biotechnology and Bioinformatics, Padmashree Dr. D. Y. Patil University, Plot No. 50, Sector 15, CBD Belapur, Navi Mumbai 400614m Maharashtra, India
20 January, 2014
04 March, 2014
05 April, 2014
01 July, 2014