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R. Sundaralingam
Vitamins Microorganisms can be used for the commercial production of certain vitamins, such as thiamine, riboflavin, folic acid, pantothenic acid, pyridoxal, vitamin B12 and Biotin. However, on a worldwide basis only the production of vitamin B12, riboflavin and ascorbic acid have major economic significance.
Vitamin B12
Vitamin B12 (Cyanocobalamin) is a vitamin that is synthesized in nature exclusively by microorganisms; because it is required by animals it is present in every animal tissue in very low concentrations (eg., 1 PPM in the liver).
The vitamin B12 needs of animals are covered by food intake or by absorption of vitamin B12 produced by intestinal microorganisms. The concentrations of vitamin B12 which are present in animal tissues are too low for use in commercial production.
Vitamin B12
Vitamin B12 was firs obtained commercially as a byproduct of streptomycete fermentations for the production of the antibiotics streptomycin, chloramphenicol or neomycin with a yield of about 1 mg/l. The most important manufacturers of vitamin B12 are: Farmitalia S.p.A (Italy); Glaxo Lab., Ltd (England); MercK & Co., Inc. (US); Rhone Poulenc S.A. (France); Roussel UCLAF (France); G. Richter Pharmaceutical Co and Chinoin (both Hungary). The current annual world production of vitamin B12 is estimated at about 12,000Kg.
In a second, aerobic phase (3-4 days) the biosynthesis of 5, 6 dimethylbenzimidazole takes place, so that 5deoxyadenosylcobalamin (Coenzyme B12) can be produced. As an alternative to the two-stage batch process in a fermenter, both stages can also be operated continuously in two tanks operated in cascade fashion.
Laboratory scale production process for Vitamin B12 using Pseudomonas denitrificans
Pseudomonas denitrificans MB 2436
Inoculum cultivation
Preculture
Production culture
5 litre Fermenter with 3.3 litre Medium c, sterilization 75 min, 120C. Inoculum: 150 ml preculture. Incubation: 90 h, 29C; stirring 420 rpm; aeration 1 vvm.
Media composition
Medium A (g / l): Sugar beet molasses 60; Yeast extract 1; N-Z-Amine 1; (NH4)2HPO4 2; MgSO4. 7H2O 1; MnSO4. H2O 0.2; ZnSO4. 7H2O 0.02; Na2MoO4. 2H2O 0.005; Agar 25; pH 7.4. Medium B (g / l): Medium A without agar. Medium C (g / l): Sugar beet molasses 100; Yeast extract 2; (NH4)2HPO4 5; MgSO4. 7H2O 3; MnSO4. H2O 0.2; Co(NO2). 6H2O 0.188; 5, 6 dimethylbenzimidazole 0.25; ZnSO4. 7H2O 0.02; Na2MoO4. 2H2O 0.005; pH 7.4
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