发酵饲料专用微生物的筛选研究
发布时间：2018-11-21 13:05
【摘要】：随着国民经济的增长,人们对食品安全性的要求不断提高,同时对环境保护和饲料安全意识也不断增强,如何解决食品安全性问题,如何寻找和开发无毒副作用、无耐药性、无蓄积残留、无环境污染的产品,应该从源头做起,目前菌体蛋白饲料产品的研发是我国饲料工业发展极为迫切需求的,也是当今社会亟待解决的问题,因此推广微生物技术的应用对于食品安全性具有重要意义。应用复合微生物发酵技术生产是对传统饲料生产模式的一种变革,同时也是现代微生物技术应用于传统生产的一种标志,当前允许应用于生产的饲用级微生物主要以芽孢杆菌、酵母菌、乳酸菌、光合细菌等四大类为主。综合查阅发现芽孢杆菌、酵母菌、乳酸菌之间具有协同互补作用并具有优良抗逆性状、耐高温、耐酸碱性、耐挤压性易贮存等独特的生物特性成为研究热点。本研究针对微生物发酵饲料现状和今后的发展趋势,以发酵食品作为微生物分离源,采用分离、鉴定并纯化的纯种乳酸菌、芽孢杆菌、酵母菌进行发酵饲料的制备研究,使之为后续规模化、工业化发酵饲料的生产奠定基础。本研究主要试验内容和结论如下:(1)从24份纳豆粉、鲜纳豆和豆豉中筛选出16株高产菌株,从20份酒曲中筛选出14株高产菌株,从20份腌制酸菜中筛选出14株高产菌株。(2)利用现代生物分析技术对上述三类菌株进行形态学和生理生化鉴定,鉴定结果表明,上述三类高产菌株分别为:N1-N16为纳豆芽孢杆菌、Z1-Z14为植物乳酸杆菌、J1-J7、J9为酿酒酵母J8、J10-J14为产朊假丝酵母。(3)将经过鉴定的菌株,通过淀粉水解和酪蛋白水解培养基初筛得到44株菌株进行淀粉酶及蛋白酶活性测定,将筛选出的最佳高产淀粉酶和蛋白酶菌株N8、Z14、J9、J14定为出发菌株,并对其生长特性进行研究,研究结果表明:N8菌株在0h-8h为迟缓期;8h-20h为对数期;20h-24h为稳定期,24h后达到衰亡期;Z14菌株在0h-2h为迟缓期,2h-14h为对数期,14h-24h为稳定期,24h后达到衰亡期;J9菌株在0h-6h为迟缓期,6h-20h为对数期,20h-30h为稳定期,30h后达到衰亡期;J14菌株在0h-8h为迟缓期,在8h-20h为对数期,20h-24h为稳定期,24h后达到期。从四菌株比较看,确定培养20h是菌种采样留菌株的最好时期。(4)经拮抗试验确定出发菌株纳豆芽孢杆菌、植物乳杆菌、酿酒酵母、产朊假丝酵母四株菌之间无抗性及抑制反应,适合进行下步固态发酵试验。(5)以蛋白含量、残渣重量为筛选指标,将未接种菌株、单一菌株与复合菌株的饲料样品进行比较,发现N8、Z14、J9、J14四株菌之间有较好的协同作用,初步筛选出用于发酵饲料最佳的纯种复合菌,为后续的研发与利用奠定了基础。
[Abstract]:With the growth of the national economy, people's requirements for food safety have been raised, and the awareness of environmental protection and feed safety has also been strengthened. How to solve the problem of food safety, how to find and develop no toxic side effects, no drug resistance, Products without accumulative residue and no environmental pollution should start from the source. At present, the research and development of bacterial protein feed products is an urgent need for the development of feed industry in our country, and is also a problem to be solved urgently in today's society. Therefore, it is of great significance to popularize the application of microbial technology for food safety. The application of compound microbial fermentation technology is a revolution to the traditional feed production mode, and it is also a symbol of the application of modern microbial technology in traditional production. At present, the forage microbes allowed to be used in production are mainly Bacillus spp. Yeast, lactic acid bacteria, photosynthetic bacteria and other four main categories. It was found that Bacillus, yeasts and lactic acid bacteria had synergistic and complementary effects and had excellent resistance to stress, high temperature, acid and alkali resistance, easy storage and so on. In view of the present situation of microbial fermentation feed and the development trend in the future, the fermentation food was used as the source of microbial isolation, and the pure lactic acid bacteria, Bacillus sp. and yeast were isolated, identified and purified to study the preparation of fermented feed. To make it for the follow-up scale, industrial fermentation feed production laid the foundation. The main contents and conclusions of this study are as follows: (1) 16 high-yielding strains were screened from 24 samples of natto powder, fresh natto and black bean, and 14 strains were screened from 20 samples of wine koji. Fourteen high-yielding strains were screened from 20 pickled vegetables. (2) morphological, physiological and biochemical identification of the three strains was carried out by modern biological analysis. The three types of high-yielding strains mentioned above are: N1-N16 is Bacillus natto, Z1-Z14 is Lactobacillus plantarum, J1-J7N J9 is Saccharomyces cerevisiae J8, J10-J14 is Candida prion. (3) the identified strains will be identified. The amylase and protease activity of 44 strains were determined by the culture medium of starch hydrolysis and casein hydrolysis. The best high yield amylase and protease strain N8Z14 / J9 / J14 was selected as the starting strain. The growth characteristics of N8 strain were studied. The results showed that N8 strain was slow in 0h-8h. 8h-20h is logarithmic phase, 20h-24h is stable phase, it reaches decaying stage after 24 hours, Z14 strain is slow phase in 0h-2h, logarithmic phase in 2h-14h phase, stable phase in 14h-24h phase, and decayed phase after 24 hours. The strain J9 was slow in 0h-6h, logarithmic in 6h-20h, stable in 20h-30h and reached the stage of death after 30 hours, while strain J14 was slow in 0h-8h, logarithmic in 8h-20h, stable in 20h-24h and reached the stage after 24 hours. From the comparison of the four strains, the best time for sampling and remaining strains was determined to be 20 hours. (4) through antagonistic test, the original strains were identified as Bacillus natto, Lactobacillus plantarum and Saccharomyces cerevisiae. The four strains of Candida prion had no resistance and inhibition reaction, so they were suitable for the next solid-state fermentation test. (5) the uninoculated strains were selected according to protein content and residue weight. By comparing the feed samples of single strain and compound strain, it was found that there was a good synergistic effect among four strains of N8, Z14, J9, J14 and J14, and the best pure compound strain for fermenting feed was screened, which laid a foundation for further research and utilization.
【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S816.3
本文编号：2347030
[Abstract]:With the growth of the national economy, people's requirements for food safety have been raised, and the awareness of environmental protection and feed safety has also been strengthened. How to solve the problem of food safety, how to find and develop no toxic side effects, no drug resistance, Products without accumulative residue and no environmental pollution should start from the source. At present, the research and development of bacterial protein feed products is an urgent need for the development of feed industry in our country, and is also a problem to be solved urgently in today's society. Therefore, it is of great significance to popularize the application of microbial technology for food safety. The application of compound microbial fermentation technology is a revolution to the traditional feed production mode, and it is also a symbol of the application of modern microbial technology in traditional production. At present, the forage microbes allowed to be used in production are mainly Bacillus spp. Yeast, lactic acid bacteria, photosynthetic bacteria and other four main categories. It was found that Bacillus, yeasts and lactic acid bacteria had synergistic and complementary effects and had excellent resistance to stress, high temperature, acid and alkali resistance, easy storage and so on. In view of the present situation of microbial fermentation feed and the development trend in the future, the fermentation food was used as the source of microbial isolation, and the pure lactic acid bacteria, Bacillus sp. and yeast were isolated, identified and purified to study the preparation of fermented feed. To make it for the follow-up scale, industrial fermentation feed production laid the foundation. The main contents and conclusions of this study are as follows: (1) 16 high-yielding strains were screened from 24 samples of natto powder, fresh natto and black bean, and 14 strains were screened from 20 samples of wine koji. Fourteen high-yielding strains were screened from 20 pickled vegetables. (2) morphological, physiological and biochemical identification of the three strains was carried out by modern biological analysis. The three types of high-yielding strains mentioned above are: N1-N16 is Bacillus natto, Z1-Z14 is Lactobacillus plantarum, J1-J7N J9 is Saccharomyces cerevisiae J8, J10-J14 is Candida prion. (3) the identified strains will be identified. The amylase and protease activity of 44 strains were determined by the culture medium of starch hydrolysis and casein hydrolysis. The best high yield amylase and protease strain N8Z14 / J9 / J14 was selected as the starting strain. The growth characteristics of N8 strain were studied. The results showed that N8 strain was slow in 0h-8h. 8h-20h is logarithmic phase, 20h-24h is stable phase, it reaches decaying stage after 24 hours, Z14 strain is slow phase in 0h-2h, logarithmic phase in 2h-14h phase, stable phase in 14h-24h phase, and decayed phase after 24 hours. The strain J9 was slow in 0h-6h, logarithmic in 6h-20h, stable in 20h-30h and reached the stage of death after 30 hours, while strain J14 was slow in 0h-8h, logarithmic in 8h-20h, stable in 20h-24h and reached the stage after 24 hours. From the comparison of the four strains, the best time for sampling and remaining strains was determined to be 20 hours. (4) through antagonistic test, the original strains were identified as Bacillus natto, Lactobacillus plantarum and Saccharomyces cerevisiae. The four strains of Candida prion had no resistance and inhibition reaction, so they were suitable for the next solid-state fermentation test. (5) the uninoculated strains were selected according to protein content and residue weight. By comparing the feed samples of single strain and compound strain, it was found that there was a good synergistic effect among four strains of N8, Z14, J9, J14 and J14, and the best pure compound strain for fermenting feed was screened, which laid a foundation for further research and utilization.
【学位授予单位】：吉林农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S816.3
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本文编号：2347030
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