Study On Sporalation Of Bacillus Coagulans

Bacillus coagulans is a kind of lactic acid bacillus, it had the very strong resistance after the spores forming, so easy of processing and storage of Bacillus coagulans as a lactic acid bacillus, this experiment is to study Bacillus coagulans fermentation of liquid and solid conditions to get much more spores.The study of the heat-resistant of spores showed that when heat at 80℃for 30min,90℃for 10min and 100℃for 10min, the survive rate was 100%. When 90℃for 30min, the survive rate was 71.6%, when 100℃for 30min, the survive rate was 59.8%, choose 80℃、10min as the conditions for viable counts.In the liquid fermentation, the first work on the basis of work had been done in the laboratory, the single factor and orthogonal tests were carried out to optimize the spore-forming condition of Bacillus coagulan in this paper. The optimal medium were confirmed as follows:Yeast extract(3g/l), peptone(5g/l), Beef extract(2g/l), MnSO4 (0.005g/l), NaCl(2g/l), K2HPO4(3g/l), MgSO4 (0.02g/l). The optimal culture were determined by single—factor test as follows:40℃, initial pH 7.0,30 ml/250 ml liquid volume, fementation for 48 h, the number of spore reached 7.6 X 108 cfu/ml.Factors promoting spore as tomato juice, corn liquid—water, and CaCO3 were promoting the spore—formation. Using Plackett-Burman design factors affecting the evaluation of the number of Bacillus. the result was glucose is negative significant, negative significant tomato juice, bran-water significantly positive, MgSO4 significantly positive. The highest climbing experiment showed that the number of spores is the amount of glucose 0.2%, tomato juice 0.8%, MgSO4 0.25%. Using response surface central composite design to optimize the significant factors. According to the analysis of spores established, get the optimal conditions for glucose 1.9 g/l, tomato juice 7.8 ml/1, corn liquid—water 10g/l, yeast extract 3g/l, K2HPO43g/l, MnSO40.3g/l, CaCO32g/l, MgSO42.4g/l, NaCl 6g/l and bran—water was used instead of distilled water. The spore value was 8.7 X 108 cfu/ml.Solid state fermentation, the optimal medium for the wheat bran medium, medium volume 15g/250ml, the optimal temperature was 45℃, and 100% inoculation.