Biochemical test and staining, salt precipitation, dialysis, de-osmosis ion- exchange chromatography, gel filtration method, SDS-PAGE.
The enzyme Glucose oxidase has significant application in therapeutics, agriculture and food industries as well as a growing impact on the bakery industry. Commonly glucose oxidase production is done from fungal source, but we made an attempt to produce this commercially important enzyme from bacterial species, isolated from soil samples which have been collected from different regions from Bangalore, India; It was identified as Pseudomonas sp. by various morphological characteristics and biochemical tests, and ken its optimum culture conditions for production as there is a little work has done on it. In general, the optimization showed the increase in enzyme activity on the basis of the minimally required incubation period of 24 hours at 40°C in alkaline pH level 10. A strain improvement was done through physical and chemical mutations to optimize the production of glucose oxidase as then proceeds to be purified through several processes and thereafter up to expectation molecular weight of the enzyme was set up by SDS-PAGE on comparison with the protein molecular weight marker.