Study on the Production of Vitamin B12 by Newly Isolated and Identified Pseudomonas aeruginosa and Effect of Gamma Irradiation on Fermentation Substrate.

Abstract

Vitamins are organic compounds that are not produced by body cells. Therefore, vitamins need to be taken from nutrients and supplements daily. They play an important role in carbohydrate, fat and protein metabolism, healthy development of the body, immunity against infections, and digestive functions. Vitamin B 12 is a necessary nutrient; it is widely used as a dietary supplement and to treat anemia all over the world. The present study focuses on isolation and extraction of vitamin B 12 producing Pseudomonas aeruginosa isolated from bladder cancer patients urine samples, the isolate was identified using morphological, physiological, biochemical methods, and confirmed by API 20 specific media, and named Pseudomonas aeruginosa M20. The vitamin B 12 production was enhanced by using Trypticase soy broth and vitamin B 12 like substance was extracted by using methanol –water as solvent. More over Two doses of Gamma irradiation were used (1 and 2 kGy) to study its effect on vitamin B 12 like substance production. The extracted vitamin B 12 like substance was characterized and identified by using HPLC, IR and EDX compared to commercial vitamin B 12 , different media (Mineral salt - methanol medium and Trypticase soy broth medium) were used for Vitamin B 12 production. In addition, different solvents (methanol –water and acetone) were used for extraction of vitamin B 12 . Trypticase soy broth and methanol –water were the most proper medium and solvent for Vitamin B 12 production with compared by commercial vitamin B 12 standard . The results presented in HPLC graphs exhibits a peak with RT at 5.422 of standard vitamin B 12 , while on Trypticase soy broth medium and methanol-water as a solvent showing a peak with RT at 5.522 which is closely related to standard Vitamin B 12 . The antimicrobial activities of both irradiated and non-irradiated bacterial extracts against bacterial and fungal strains were tested and two doses of Gamma irradiation were used (1 and 2 kGy) to study the effect of Gamma irradiation on the anti-microbial activity of the extract against the bacterial and fungal strains.