Abstract

PURIFICATION AND CHARACTERIZATION OF CYCLODEXTRIN GLYCOSYLTRANSFERASE FROM ALKALIPHILIC BACILLUS OSHIMENSIS

Reshma Nilesh Pol and *Arpita Mandar Gupte

Abstract

Alkaliphilic cyclodextrin glycosyltransferase (CGTase) producer was isolated from the rice fields of Maharashtra, India. 16S rRNA sequence analysis identified the isolate as Bacillus oshimensis. CGTase enzyme was purified by ammonium sulphate precipitation along with starch adsorption, which is a simple and efficient method followed by ultra filtration. The purity was found to increase by 2.5 fold with 11% yield. The molecular weight of partially purified enzyme was found to be 66kDa respectively, as determined by Native poly-acrylamide gel electrophoresis. The characterization of the enzyme was carried out for pH and temperature and was found to be optimum at pH 8 and 37oC. Km was found to be 1.66 g% and Vmax as 19.38 U/ml.

Keywords: Cyclodextrin glycosyltransferase, purification, characterization.