| Bacillus natto and Lacto.acidophilus were used to ferment the solid-state bee pollen, the optimum strains and optimum conditions of pollen fermentation were determined, the change of all the main ingredients and active substance in the bee pollen after the fermentation was analyzed, at the same time, the health function of the fermentation bee pollen was also studied by the means of animal experiment. The conditions in the lab was finished, which became the base of producing a new style fermentation pollen which owned not only probiotic characters but natural pollen functions and wonderful odor and taste. The main contents and the results of the study were as follows:1.Lactic acid bacteria(Lactobacillus bulgaricus,Streptococcus thermophilus, L.acidophilus, L.plantarum), Bacillus natto and yeast were used to ferment solid-state pollen by single bacteria and mixed bacteria, at last, the optimum strains and optimum conditions of pollen fermentation were determined:fermentation mode:superficial aerobic fermentation;inoculum mode: when ferment pollen, first, inoculate Bacillus natto, second, after 3 days, inoculate L.acidophilus for 5d;optimum inoculum volume:10%;optimum moisture content:35%～40%;fermentation temperature:30℃ ,fermentation period:8～9d.On the base of this, the fermentation conditions of the bee pollen were determined in the 20L fermentor:volume of the bee pollen 20%, fermentation temperature:30℃,shaker spacing time:2h,shaker time:0.5h, fermentor pressure:0.03～0.05MP,after inoculate L.acidophilus, aerating spacing time:12h, aerating time:2h, moisture content:35%～40%, inoculum volume: 10%, fermentation period:8～9d. The product had wonderful odor and taste, the shortcoming of natural bee pollen is overcomed effectively. The live count of Bacillus natto is 4.5×10~7cfu/g in the product and the live count of L.acidophilus is 2.7×10~7cfu/g in the product.2.The change of all the main ingredients and active substance in the bee pollen after the fermentation was analyzed. After the fermentation, the coarse-protein increased 5.9%,the coarse-fat decreased 1.57%, free amino acid increased 0.02%,organic acid increased 0.79g/100g,pH decreased to 4.2,the lactic acid increased 0.20g/100g and was the most of all organic acids. the protease andnattokinase activity in the bee pollen were the highest respectively 1023.5u/g for Id and 5.78IU/g for 4d.3.The change of the peptides and small molecule sensitized protein in the bee pollen during the fermentation was analyzed. Compared the peptides of fermented pollen with the ones of natural pollen by RP—HPLC ,three peptides in fermented pollen disappeared. The change of taste of fermented pollen was probably connected with this, however, the more experiments are needed to make;it was proved that the small molecule sensitized protein range from 20kD to 40kD disappeared in the fermented pollen by SDS — PAGE, similarly, the more experiments are also needed to make.4.The health function of the fermentation bee pollen was studied by the means of animal experiment.Firstly,there was no significantly difference about enforcing the spleen of mice between the fermentation bee pollen and the natural bee pollen;the fermentation bee pollen had more stimulative effect on the thymus of mice than the natural bee pollen on the condition of high dosage,while there was no significantly difference between them on the condition of middle and low dosage, however, in any case, they were obviously stronger than control group;comparing the fermentation bee pollen with the natural bee pollen, the former increased HC50 of normal mice on the condition of high dosage, but the both had no significantly difference on the condition of middle and low dosage;as for the effect assayed by MTT of the bee pollen on splenocyte proliferation responses induced by ConA,the result was the same with the one of HCso.In a word, the fermentation bee pollen were able to promote further immunologic function of the body.
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