饲用微生态制剂固态发酵工艺的初步研究
发布时间：2018-08-25 07:11
【摘要】：微生态制剂的固态发酵比液体发酵能提供更为丰富、活性更高的酶系,其所有发酵产物均可作为制剂组分,且具有能耗低、无废液排放等优势。因此本文首先对饲用微生态制剂进行菌株的筛选和菌株种子液培养基的优化,然后初步建立了一套饲用微生态制剂的固态发酵工艺,并对菌株种子液和制剂的安全性进行验证。其结果如下:(1)通过对干酪乳杆菌M9(Lactobacillus casei M9)进行耐酸、耐猪胆盐和耐高温驯化及对产朊假丝酵母Y5(Candida utilis Y5)进行耐高温驯化以提高两株供试菌株的耐受性并得到新的菌株:干酪乳杆菌M9c(Lactobacillus casei M9c)和产朊假丝酵母Y5a(Candida utilis Y5a);然后对枯草芽孢杆菌R5(Bacillus subtilis R5)、干酪乳杆菌M9c、产朊假丝酵母Y5a进行菌株间拮抗作用确定各自之间不存在拮抗作用,得到可以研制饲用微生态制剂的优良菌株。(2)通过单因素试验及正交试验,得到三种菌的最优培养基及培养条件:B.subtilis R5:葡萄糖2%,蛋白胨1%,NH4Cl 0.6%,MgSO4 0.6%,NaCl 0.5%,pH 6.5。接种量为5%,34℃,180rpm摇床培养16h。L.casei M9c:酵母膏1%,蛋白胨2%,红糖2%,NH4Cl 0.6%,KH2PO4 0.4%,MgSO4 0.2%。pH 6,接种量为2.5%,37℃静置培养16h。C.utilis Y5a:去皮马铃薯20%,葡萄糖1%,酵母膏0.5%,KH2PO4 0.05%,ZnSO40.01%,MnSO4 0.075%。pH为6,接种量为5%,32℃,180rpm摇床培养16h。(3)通过PB试验挑选出培养基组分的三个关键因素,采用最陡爬坡实验确定其最佳浓度区域,利用中心旋转组合试验确定固态发酵培养基的配比:麦麸18g,玉米皮粉7.5g,豆粕4.5g,麦芽糖0.45g,红糖0.74g,氯化铵0.3g,硫酸铵0.3g,碳酸钙0.15g,磷酸二氢钾0.29g,硫酸锰0.08,硫酸镁0.15。由单因素试验确定了固态发酵条件为:含水量(即干物料质量:水的体积)为1:0.8,B.subtilis R5、L.casei M9c、C.utilis Y5a的接种量分别为5%、2.5%、5%,整个发酵过程中温度控制在30℃~34℃,发酵44h。(4)通过对发酵产物进行一系列处理,初步得到一套发酵后预处理工艺及保存方式:将发酵产物置于60℃真空干燥箱烘至含水量为10%,置于18×25全新PET/PE复合加工而成真空袋中4℃真空保藏,180d后活菌数依然可达1.033×1011 cfu/g,高于国家标准。(5)用筛选的菌株种子液和固态发酵的微生态制剂灌喂小白鼠14~30天,小白鼠均未出现死亡情况,并且小白鼠饮食、精神面貌等临床现象都正常。解剖观察小白鼠的肝脏和肾脏,未发现有病变等不良现象,并且制剂可有效增加小白鼠的体重,降低腹泻率。
[Abstract]:The solid-state fermentation of microecological agents can provide more enzymes with higher activity than liquid fermentation. All the fermentation products can be used as preparation components and have the advantages of low energy consumption and no waste liquid emission. Therefore, in this paper, the strains were screened and the medium of seed liquid was optimized, then a set of solid-state fermentation technology was established, and the safety of the seed liquid and preparation was verified. The results are as follows: (1) the acid tolerance of Lactobacillus casei M9 (Lactobacillus casei M9 was studied. High temperature tolerance of Candida utilis Y5 (Candida utilis Y5) in order to improve the tolerance of the two strains and to obtain new strains: Lactobacillus casei M9c (Lactobacillus casei M9c) and Candida utilis Y5a (Candida utilis Y5a). Bacillus subtilis R5 (Bacillus subtilis R5), Lactobacillus casei M9c, Candida prion Y5a were used for the antagonism between strains. The optimum culture medium and culture conditions of the three strains were obtained by single factor test and orthogonal test. The optimum culture conditions were: glucose 2, peptone 1 + NH 4Cl 0.6 + MgSO 4 0. 6% NaCl 0.5 + pH 6. 5. The optimum medium and culture conditions of the three strains were obtained by single factor test and orthogonal experiment. The optimum culture medium and culture conditions were as follows: 1% B. subtilis R5: glucose 2%, peptone 1% NH 4 Cl 0. 6 and MgSO 4 0. 6%. The inoculation amount was 5 ~ 34 鈩，
本文编号：2202127
[Abstract]:The solid-state fermentation of microecological agents can provide more enzymes with higher activity than liquid fermentation. All the fermentation products can be used as preparation components and have the advantages of low energy consumption and no waste liquid emission. Therefore, in this paper, the strains were screened and the medium of seed liquid was optimized, then a set of solid-state fermentation technology was established, and the safety of the seed liquid and preparation was verified. The results are as follows: (1) the acid tolerance of Lactobacillus casei M9 (Lactobacillus casei M9 was studied. High temperature tolerance of Candida utilis Y5 (Candida utilis Y5) in order to improve the tolerance of the two strains and to obtain new strains: Lactobacillus casei M9c (Lactobacillus casei M9c) and Candida utilis Y5a (Candida utilis Y5a). Bacillus subtilis R5 (Bacillus subtilis R5), Lactobacillus casei M9c, Candida prion Y5a were used for the antagonism between strains. The optimum culture medium and culture conditions of the three strains were obtained by single factor test and orthogonal test. The optimum culture conditions were: glucose 2, peptone 1 + NH 4Cl 0.6 + MgSO 4 0. 6% NaCl 0.5 + pH 6. 5. The optimum medium and culture conditions of the three strains were obtained by single factor test and orthogonal experiment. The optimum culture medium and culture conditions were as follows: 1% B. subtilis R5: glucose 2%, peptone 1% NH 4 Cl 0. 6 and MgSO 4 0. 6%. The inoculation amount was 5 ~ 34 鈩，
本文编号：2202127
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