Research On Cleaned Fermentation Medium Of Clostridium Butyricum

Clostridium butyricum, 1933 by Dr. kingi Miyairi first found in human and animal existence in an anaerobic intestinal probiotics. It can formation enspore, and have strong ability to resist high temperatures, low pH, bile concentrations and some antibiotics. Clostridium butyricum in Japan and other countries as widespread human gastrointestinal function conditioning medicinal drug use, can significantly reduce the pseudomembranous colitis the incidence of dysbacteriosis caused by diarrhea, antibiotic-associated colitis, constipa-tion, etc, have a good effect, and without any side effects. Its application in feeds and veterinary drug has a very vast prospect compared to non-spore formation probiotics.It is hard to storage Clostridium butyricum directly, but is easy to be coated or be hold under low temperature. And using the way of coating and low temperature with high concentration strain powder was more economical. So in this paper, a cleared fermentation strategy was taken to obtain high-content products.In order to enhance the biomass production by Clostridium butyricum B1, this experiment has carried on the optimization to its cleaned fermentation medium. Initially, the most suitable carbon glucose, nitrogen source trypone, yeast extract, and beef extract were selected according to single factorial experiment respectively.Based on the result, two level factorial design of Plackett-Burman was used to evaluate the influence factors. It shows that three factors played the important roles in the medium, it was glucose, trypone and yeast extract. Then the path of Steepest Ascent was used to approach the optimal region. The optimal combined concentration for maximum biomass production were further optimized by response surface methodology and determined as follows: glucose 3.4 %, trypone 4.0 %, yeast extract 3.8 %. The optimal concentrations of the variables were determined as: glucose 3.4%(w/v) , trypone 4%(w/v) , yeast extract 3.8%(w/v) ,beef extract 1.5% (w/v) ,starch 0.1%(w/v) , NaCl 0.5%(w/v) , CH₃COONa 0.3%(w/v) , cys 0.05% (w/v) ,K₂HPO₄ 0.2%(w/v) ,(NH₄)₂SO₄ 0.1%(w/v) ,original pH 7.1 .The optimization of culture medium of Clostridium butyricum B1,the biomass production was improved from 6.96×10 cfu/mL to 3×10 cfu/mL , enhanced 4.3 folds, and the number of each gram dry matter is 5.57×10¹⁰ cfu.