Establishment Of RP-HPLC For Biotin Content And Effection Of The Pimelic Acid On Biotin Production

Biotin is one of essential vitamins for animals during their growth process because the nutrition is playing an ever more significant role in the normal growth, breeding, feed utilization and developments of skin, bones and feather of animals. So far the biotin is produced by chemical methods mainly. Some scientists are trying to produce more biotin by microorganism due to some drawbacks of chemical synthesis methods such as complex interaction, high-cost, continual pollution. Some achievements in scientific research summed up by our predecessors showed that the biotin not only be can synthesized by subtilis, fungi, and other microorganisms , but their production cost is much lower than that by chemical methods and the produce process is adaptable to factory industrial production. On the basis of the advanced experience and technology of predecessors, The bacillus BQJ-34 which can synthesized biotin was chosen and was cultured. The level of the carbon source needed by the bacillus and the pimelic acid as a precursors for biotin is adjusted to enhance production of the biotin.BQJ-34 was cultured on the biotin fermentation media for 48 hour ,a 10ml fluid medium was boiled for an hour and was centrifuged (10,000r/min , 30 min), the supernatant is filleted with the 0.22μm filters and the biotin content of filter liquid is detected by RP-HPLC based on the biological samples regression analysis, recovery, accuracy testing. It was affirmed that the HPLC is useful for determination of the biotin in fermentation system eventually. And the best testing conditions is that an Anjielun Zorbax SB-C18 (4.6 issue 250cm,5μm) column is used , Volume Ratio (a mibile phase) of K₂HPO₄ (0.04mol, pH2.5)and CH₃CN is 91.5:8.5, and that the ultraviolet detection wavelength is 215nm.In order to determine the optimal additional level of maltose in the fermentation medium, the medium was divided into six groups and maltose were added to the medium so that the content of maltose were 0%,2%,4%,6%,8% and 10% separately. Then the BQJ-34 were added to the medium shocking under 220r/min conditions to culture them at 37℃. The liquid absorption value and growth of the bacillus was measured after 24, 48 and 72 hour. It was showed that the most effective level of maltose in the medium is 5%.The media with pimelic acid was centrifugated (30min at 10000r/min, room temperature )...