Abstract

A STUDY ON CAPABILITY OF ? (BETA) - GLUCOSIDASE GENE TO BE CLONED INTO ESCHERICHIA COLI-DH5? ISOLATED FROM BIFIDOBACTERIUM BREVE

*Sharmistha Pal and Madhulika Singh

ABSTRACT

The research comprises isolation and identification of Bifidobacterium strains from Amul Flaavyo Probiotic Yoghurt and Amul Lactose Free Milk. Bifidobacterium Agar media, containing Bifidobacterium Selective Supplement (Propionic acid) or stimulating substances (lactulose instead of lactose), and anaerobic culture conditions were used for strain isolation. Identification of isolates to the genus was based on fructose-6-phosphate phosphoketolase activity, breakdown of urea in to ammonia, use butylenes glycol pathway to metabolize pyruvic acid determined by methyl red test, lack of ability to produce catalase, indole from tryptophan, digestion of glucose to acetylmethylcarbinol determined by voges proskeaur test, gas from glucose, and unable to utilize citrate. The sole purpose of this project was to clone β-Glucosidase gene of Bifidobacterium into a suitable vector (pUC18) and to transform this vector into a host (E.coli DH5α). The formation of recombinant cell was verified by growing in Luria Bertini Ampicillin agar plates and also the β-Glucosidase enzyme assay was carried using pNPG out for final check. The significance of Bifidobacterium and its β-Glucosidase enzyme in the food, medical and pharmaceutical industry is no more concealed. So, there is an urgent need to find an alternative for the cost effective large scale production of β-Glucosidase which is demonstrated in this project.

Keywords: ?-Glucosidase Gene, Bifidobacterium, Cloning, Enzyme Activity, Probiotics.