Research On The Extraction Process Of Adenosylcobalamin Produced By Pseudomonas Denitrificans

Vitamin B₁₂ is widely used in medicine stockbreeding and food additive,which is produced mainly by fermentation. In this paper, a new method to determine vitamin B₁₂ in industrial fermentation was studied.VB₁₂ extraction from fermentation broth was also researched.The study in this paper was based on industy applications,which have special valuable reference to improvement of VB₁₂ fermentation and extraction.1.Method to determine VB₁₂ in fermentation brothVitamin B₁₂ is produced by fermentation. Adenosylcobalamin are mainly products by fermentation.In this paper,VB₁₂ was produced by Pseudomonas denitrificans fermentation, cells were collected by centrifugation.Sample was analyzed by HPLC after pretreatment. Sample can be well separated by themethod in 25min.The data all meet the needs of methodology.The method is simple,rapid and accurate,which can be used in the and extraction process of VitaminB₁₂.2.Four methods to extract VB₁₂ from Pseudomonas denitrificans by cell disruptionFour methods were applied to disrupt cell for extracting VB₁₂ , including ultrasonication boiling,microwave heating,high-pressure homogenization.The kinetics of cell disruption was studied.Experiment showed that methods of boiling and microwave heating can get high extraction ratio,and protein released from cell can be removed in operating process.The method of boiling is suitable for industry production,because it needs simple equipment and can be managed in large scale.3.Separation and purification adenosylcobalamin from fermentation broth by macroporous adsorption resinsFour different macroporous adsorption resins were studied for research the processes of static adsorption and desorption for adenosylcobalamin.Mean while influence of PH and temperature on adsorption and desorption were determined.The type of 3# resin has excellent performance in static adsorption and desorption for adenosylcobalamin.Optimal dynamic conditions were investigated for 3# resin in process of adsorption and desorption,and kinetic of which was discussed.As a result,aprimaryprocess for separation and purification adenosylcobalamin from fermentation broth using 3# resins was established.4.Qualitative Analysis Using the former manufacturing experience of Vitamin B₁₂,the structure analysis, quality analysis and stability analysis of pure adenosylcobalamin were studied. In the end, the article confirm that adenosylcobalamin passing through several process comply the request, the product quality accord with the request of the state pharmacopoeia.