| Solid-state fermentation of micro-ecological agent can provide richer,more active enzymes than the liquid fermentation.All the fermentation products are also used as formulation components,which shares with the advantages such as low energy consumption,non-waste emissions and so on.This paper firstly had screened strains of micro-ecological agent and the optimization of strain seed liquid culture medium.Then there was a preliminary set up of the process of solid state fermentation on micro-ecological agent,and the verification of security on liquid seed strains and micro-ecological agent.Test results are as follows:(1)Lactobacillus casei M9 were processed with the test of acids resistance,pig bile resistance and temperature resistance,in addition,Candida utilis Y5 was conducted with high temperature acclimation to increase their tolerance and got the new strains: Lactobacillus casei M9 c and Candida utilis Y5 a.Then through the antagonism test among Bacillus subtilis R5?Lactobacillus casei M9c?Candida utilis Y5 a to make sure that the inhibition does not exist among each other.(2)From single factor test and orthogonal experiment,there got three bacteria's optimal medium and culture conditions.B.subtilis R5: glucose 2%,peptone 1%,NH4 Cl 0.6%,MgSO4 0.6%,NaCl 0.5%,pH 6.5.Inoculation of 5%,34?,180 rpm shaker cultivated16 h.L.casei M9c: yeast extract 1%,peptone 2%,brown sugar 2%,NH4 Cl 0.6%,KH2PO4 0.4%,MgSO4 0.2%.pH 6.Inoculum was 2.5%,37? static culture 16 h.C.utilis Y5a: peeled potatoes 20%,glucose 1%,yeast extract 0.5%,KH2PO4 0.05%,ZnSO4 0.01%,MnSO4 0.075%.pH 6,inoculation of 5%,32?,180 rpm shaker cultivated16 h.(3)Through Plackett-Burman experiment to pick out three key factors of medium components,and to make sure of the optimal concentration region through steepest ascent experiments.To ensure the ratio of the fermentation medium through center rotation combination test: wheat bran 18 g,corn bran powder 7.5g,soybean 4.5g,maltose 0.45 g,brown sugar 0.74 g,ammonium sulfide 0.3g,ammonium sulfate 0.3g,calcium carbonate 0.15 g,potassium dihydrogen phosphate 0.29 g,manganese sulfate 0.08 g,magnesium sulfate 0.15 g.Fermentation conditions were optimized by single factor experiment: water content(ie dry material quality: the volume of water)is 1: 0.8,inoculums of Bacillus subtilis R5?Lactobacillus casei M9 c and Candida utilis Y5 a was 5%?2.5%?5% respectly,throughout the fermentation temperature was controlled at 30?~34?for 44 hours.(4)A preliminary understanding of the pretreatment and Storage ways were obtained after fermentation through a series of process to fermentation material: The fermented material was placed in a vacuum drying oven under the temperature of 60?with a moisture content to 10%,then placed in 18×25 brand new PET / PE composite processing vacuum bag at 4?vacuum preservation,after 180 d,viable count still remains 1.033×1011cfu/g,higher than the national standard.(5)when fed the mice with seed inoculant and fungicide for 14days~30days,there is no death phenomenon.In addition,diet of mice and mental outlook and so on are normal.Through the anatomy of mice's liver and kidney,no lesion and other undesirable phenomena were found.Besides,agents can effectively increase the body weight of mice,and reduce diarrhea rates. |
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