发酵西兰花渣生产动物饲料的菌种筛选及其发酵工艺的研究

发布时间：2018-03-02 18:20

  本文选题：西兰花渣　切入点：发酵饲料　出处：《内蒙古农业大学》2017年硕士论文　论文类型：学位论文

【摘要】：西兰花又名绿菜花,是十字花科芸薹属甘蓝种中的一个变种,富含蛋白质、碳水化合物、矿物质、维生素等多种营养物质。每到收获季节约占西兰花植株的40-50%的西兰花副产物(茎、叶、残次品)被遗弃在田间,本文以西兰花渣为主要原料进行动物饲料生产的工艺研究,为西兰花综合利用提供一条经济合理的利用之路,具体实验结果如下:1.利用酪蛋白培养基由西兰花的田间土壤、残基、污泥等3份样品中筛选出30株芽孢杆菌,并通过酪蛋白培养基和纤维素培养基进行点种实验从中筛选出降解蛋白和纤维素能力较强的一株芽孢杆菌,菌株代号为CTL-25;利用含2.0%的无水碳酸钙的MRS固体培养基从西兰花菜叶样品中筛选出一株产酸能力较强的乳酸菌,菌株代号为CTM-1。经16S rDNA基因序列比对和同源性分析得知,菌株CTL-25和菌株CTM-1分别为枯草芽孢杆菌和屎肠球菌。2.将枯草芽孢杆菌CTL-25、屎肠球菌CTM-1、酵母菌CTJ-1(CT组合菌)按1:1:1的比例、2.52×106 CFU/g的接种量接种于含西兰花渣50.0%、喷浆玉米皮10.0%、米糠粕20.0%、麸皮20.0%组成的饲料配方中发酵,发酵结束后以饲料中酸溶蛋白含量为参考指标得知,CT组合菌发酵的西兰花渣饲料的酸溶蛋白含量为5.19%,而市场上购买的四份发酵剂发酵的西兰花渣饲料中的酸溶蛋白含量分别为4.98%、5.91%、4.89%、5.29%。说明CT组合菌发酵对西兰花渣发酵饲料,提高其酸溶蛋白含量具有很好的应用潜力。3.通过将CT组合菌以1:1:1的比例、5.04×106 CFU/g的接种量接种于3种不同的西兰花渣配比的配方中得知,由西兰花渣50.0%、米糠粕20.0%、麸皮20.0%、喷浆玉米皮10.0%组成的饲料配方中的酸溶蛋白含量最高,即5.56%。然后将CT组合菌以2.52×106CFU/g的接种量进行单菌、双菌、三菌发酵实验,其中双菌比例为1:1、三菌比例为1:1:1,实验结果表明与单菌、双菌发酵相比利用三菌发酵不仅增加了西兰花渣发酵饲料中益生菌种类,而且饲料中酸溶蛋白含量显著高于其他两种菌处理组(p0.05),通过单因素实验和正交实验确定最佳的菌种配比为;菌株CTM-1:菌株CTJ-1:菌株 CTL-25=1:14:6。4.通过单因素实验和正交实验得知,利用西兰花渣生产动物饲料的最佳发酵工艺为:菌株CTM-1、菌株CTJ-1、菌株CTL-25的配比为1:14:6,接种量为7.5×106 CFU/g、发酵时间为3d、含水量为47.0%、发酵温度为31℃,此时发酵饲料中酸溶蛋白含量为6.68%、粗蛋白含量为23.50%、pH值为4.5、还原糖含量为3.0%,与对照组(未添加发酵剂)相比酸溶蛋白含量提高150.0%、粗蛋白含量提高15.0%、还原糖含量提高50.0%。
[Abstract]:Broccoli, also known as cauliflower, is a variety of Brassica species in the cruciferous family, rich in protein, carbohydrates, minerals, The by-products of broccoli (stems, leaves, residues) which account for 40-50% of broccoli plants in harvest season are abandoned in the field. In this paper, the production process of animal feed with broccoli dregs as the main raw material is studied. The experimental results are as follows: 1. Using casein medium, 30 strains of Bacillus spp were screened from soil, residue, sludge and other three samples of broccoli. A strain of Bacillus with strong ability to degrade protein and cellulose was screened from casein medium and cellulose medium. The strain was codenamed CTL-25, and a lactic acid bacteria with strong acid-producing ability was screened from broccoli leaves on MRS solid medium containing 2.0% anhydrous calcium carbonate. The strain was code-named CTM-1.The results of 16s rDNA gene sequence alignment and homology analysis showed that the lactic acid bacteria had strong acid-producing ability. Strain CTL-25 and strain CTM-1 were Bacillus subtilis and Enterococcus faecium respectively. 2.The inoculation amount of Bacillus subtilis CTL-25, Enterobacter faecium CTM-1 and yeast CTJ-1(CT was 2.52 脳 10 ~ 6 CFU/g in proportion of 1: 1: 1, respectively. Bran meal 20. 0%, wheat bran 20.0% composition of the feed formula fermentation, At the end of fermentation, the acid soluble protein content of broccoli dregs fermented by CT was found to be 5.19, while the acid soluble egg in the four starter fermented broccoli residue feed was purchased on the market. The white content was 4.98% 5.91% and 4.89% 5.29%, respectively. It has a good application potential to increase the content of acidolytic protein. 3.The results showed that the inoculation amount of CT combination bacteria was 5. 04 脳 10 ~ 6 CFU/g at the ratio of 1: 1: 1: 1 in three different formulations of broccoli residue. The content of acid-soluble protein in the feed formula composed of broccoli residue 50.0g, rice bran meal 20.0 and bran 20.0 was the highest in the feed formula composed of spraying corn peel 10.0%, that is 5.560.Then, the fermentation experiments of single, double and triple bacteria were carried out with the inoculation amount of 2.52 脳 10 6 CFU / g for CT combination bacteria. The ratio of two bacteria is 1: 1, and the proportion of three bacteria is 1: 1: 1. The experimental results show that compared with single bacteria and double bacteria fermentation, the use of three bacteria fermentation not only increases the variety of probiotics in the fermented feed of broccoli dregs. Moreover, the content of acid-soluble protein in feed was significantly higher than that in the other two bacteria treatment groups, and the optimum proportion of strains CTM-1: CTJ-1: strain CTL-251: 14: 6.4was determined by single factor experiment and orthogonal experiment. The results of single factor experiment and orthogonal experiment showed that, The optimum fermentation process for producing animal feed from broccoli residue was as follows: strain CTM-1, strain CTJ-1, proportion of strain CTL-25 was 1: 14: 6, inoculation amount was 7.5 脳 10 ~ 6 CFU / g, fermentation time was 3 days, water content was 47.0 鈩，

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