發(fā)酵飼料專用微生物的篩選研究
發(fā)布時間：2018-11-21 13:05
【摘要】：隨著國民經(jīng)濟的增長,人們對食品安全性的要求不斷提高,同時對環(huán)境保護和飼料安全意識也不斷增強,如何解決食品安全性問題,如何尋找和開發(fā)無毒副作用、無耐藥性、無蓄積殘留、無環(huán)境污染的產(chǎn)品,應(yīng)該從源頭做起,目前菌體蛋白飼料產(chǎn)品的研發(fā)是我國飼料工業(yè)發(fā)展極為迫切需求的,也是當(dāng)今社會亟待解決的問題,因此推廣微生物技術(shù)的應(yīng)用對于食品安全性具有重要意義。應(yīng)用復(fù)合微生物發(fā)酵技術(shù)生產(chǎn)是對傳統(tǒng)飼料生產(chǎn)模式的一種變革,同時也是現(xiàn)代微生物技術(shù)應(yīng)用于傳統(tǒng)生產(chǎn)的一種標志,當(dāng)前允許應(yīng)用于生產(chǎn)的飼用級微生物主要以芽孢桿菌、酵母菌、乳酸菌、光合細菌等四大類為主。綜合查閱發(fā)現(xiàn)芽孢桿菌、酵母菌、乳酸菌之間具有協(xié)同互補作用并具有優(yōu)良抗逆性狀、耐高溫、耐酸堿性、耐擠壓性易貯存等獨特的生物特性成為研究熱點。本研究針對微生物發(fā)酵飼料現(xiàn)狀和今后的發(fā)展趨勢,以發(fā)酵食品作為微生物分離源,采用分離、鑒定并純化的純種乳酸菌、芽孢桿菌、酵母菌進行發(fā)酵飼料的制備研究,使之為后續(xù)規(guī)�；�、工業(yè)化發(fā)酵飼料的生產(chǎn)奠定基礎(chǔ)。本研究主要試驗內(nèi)容和結(jié)論如下:(1)從24份納豆粉、鮮納豆和豆豉中篩選出16株高產(chǎn)菌株,從20份酒曲中篩選出14株高產(chǎn)菌株,從20份腌制酸菜中篩選出14株高產(chǎn)菌株。(2)利用現(xiàn)代生物分析技術(shù)對上述三類菌株進行形態(tài)學(xué)和生理生化鑒定,鑒定結(jié)果表明,上述三類高產(chǎn)菌株分別為:N1-N16為納豆芽孢桿菌、Z1-Z14為植物乳酸桿菌、J1-J7、J9為釀酒酵母J8、J10-J14為產(chǎn)朊假絲酵母。(3)將經(jīng)過鑒定的菌株,通過淀粉水解和酪蛋白水解培養(yǎng)基初篩得到44株菌株進行淀粉酶及蛋白酶活性測定,將篩選出的最佳高產(chǎn)淀粉酶和蛋白酶菌株N8、Z14、J9、J14定為出發(fā)菌株,并對其生長特性進行研究,研究結(jié)果表明:N8菌株在0h-8h為遲緩期;8h-20h為對數(shù)期;20h-24h為穩(wěn)定期,24h后達到衰亡期;Z14菌株在0h-2h為遲緩期,2h-14h為對數(shù)期,14h-24h為穩(wěn)定期,24h后達到衰亡期;J9菌株在0h-6h為遲緩期,6h-20h為對數(shù)期,20h-30h為穩(wěn)定期,30h后達到衰亡期;J14菌株在0h-8h為遲緩期,在8h-20h為對數(shù)期,20h-24h為穩(wěn)定期,24h后達到期。從四菌株比較看,確定培養(yǎng)20h是菌種采樣留菌株的最好時期。(4)經(jīng)拮抗試驗確定出發(fā)菌株納豆芽孢桿菌、植物乳桿菌、釀酒酵母、產(chǎn)朊假絲酵母四株菌之間無抗性及抑制反應(yīng),適合進行下步固態(tài)發(fā)酵試驗。(5)以蛋白含量、殘渣重量為篩選指標,將未接種菌株、單一菌株與復(fù)合菌株的飼料樣品進行比較,發(fā)現(xiàn)N8、Z14、J9、J14四株菌之間有較好的協(xié)同作用,初步篩選出用于發(fā)酵飼料最佳的純種復(fù)合菌,為后續(xù)的研發(fā)與利用奠定了基礎(chǔ)。
[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.
【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S816.3
本文編號：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.
【學(xué)位授予單位】：吉林農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：S816.3
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