發(fā)布時(shí)間：2018-02-12 21:20

  本文關(guān)鍵詞： 枯草芽孢桿菌 芽孢 發(fā)酵工藝 微生物菌劑 制備　出處：《河北科技大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：腐熟菌劑主要是指針對(duì)畜禽糞便,植物殘?bào)w以及一些生活廢棄物等難分解的有機(jī)物進(jìn)行處理而研究生產(chǎn)出的一類微生物菌劑,廣泛應(yīng)用農(nóng)作物秸稈、畜禽糞便的資源化和無害化處理。枯草芽孢桿菌是在自然界廣泛存在的一種好氧的產(chǎn)芽孢桿菌。它可以產(chǎn)生蛋白酶、淀粉酶、纖維素酶和半纖維素酶等酶,是農(nóng)作物秸稈、畜禽糞便等有機(jī)物料腐熟菌劑的主要菌種之一。芽孢數(shù)和有效活菌數(shù)是腐熟菌劑質(zhì)量的重要指標(biāo)。生產(chǎn)成本也是微生物菌劑生產(chǎn)企業(yè)所關(guān)心的重要問題。本課題研究了枯草芽孢桿菌DK36的產(chǎn)芽孢條件,優(yōu)化了發(fā)酵工藝。在此基礎(chǔ)上,研究了菌劑的制備工藝。為了降低生產(chǎn)成本,進(jìn)行了以脫酚棉籽蛋白為氮源培養(yǎng)枯草芽孢桿菌DK36的研究。其主要研究結(jié)果如下:1)枯草芽孢桿菌DK36的產(chǎn)芽孢條件研究。當(dāng)葡萄糖、蔗糖、果糖、可溶性淀粉、玉米粉、麩皮為碳源時(shí),菌體生長(zhǎng)良好。葡萄糖和蔗糖作碳源時(shí)均不產(chǎn)生芽孢。當(dāng)麩皮作為碳源時(shí),形成的芽孢最多。其次為玉米粉、可溶性淀粉。蛋白胨、酵母浸粉、大豆蛋白胨、豆餅粉、硝酸鉀、硝酸鈉、硝酸銨均可使菌體大量形成芽孢,其中大豆蛋白胨的效果最好。尿素抑制菌體生長(zhǎng)和芽孢形成。添加Ca2+、Mg2+、Na+、Fe2+、Mn2+均可促進(jìn)芽孢的形成,其中效果最明顯的是硫酸錳。Cu2+對(duì)生長(zhǎng)和形成芽孢均有很大抑制作用。產(chǎn)芽孢最佳培養(yǎng)基:麩皮15.0 g/L、大豆蛋白胨5.0g/L、硫酸錳0.1 g/L、碳酸鈣3.0 g/L。最佳產(chǎn)芽孢發(fā)酵條件:培養(yǎng)溫度30℃、轉(zhuǎn)速200 r/min、接種為2%、培養(yǎng)基初始pH為7.0,芽孢數(shù)達(dá)3.6×109 cfu/mL。2)脫酚棉籽蛋白培養(yǎng)枯草芽孢桿菌DK36的研究。棉籽蛋白作為氮源時(shí),菌體生長(zhǎng)良好。碳源的種類對(duì)產(chǎn)芽孢有顯著影響。葡萄糖、蔗糖、玉米粉、麩皮均使菌體生長(zhǎng)良好。但葡萄糖和蔗糖為碳源,菌體不形成芽孢,而玉米粉和麩皮可使菌體產(chǎn)生大量芽孢。添加Ca2+、Mn2+、Mg2+和K+對(duì)芽孢形成有明顯的促進(jìn)作用,并且發(fā)現(xiàn)鉀鹽與鎂鹽、錳鹽、鈣鹽復(fù)合效果更佳。研究還發(fā)現(xiàn),脫酚棉籽蛋白作為氮源會(huì)造成芽孢形成緩慢。脫酚棉籽蛋白培養(yǎng)枯草芽孢桿菌的最佳培養(yǎng)基:玉米粉30.0g/L、棉籽蛋白30.0 g/L、硫酸鎂2.0 g/L、磷酸二氫鉀4.5 g/L。其最佳發(fā)酵條件:培養(yǎng)溫度30℃、搖床轉(zhuǎn)速200 r/min、接種量為8%,芽孢數(shù)和芽孢率分別達(dá)到3.5×109cfu/mL和98.7%。3)枯草芽孢桿菌DK36菌劑的制備。盡管芽孢能耐高溫,但隨著干燥溫度升高,菌劑有效活菌數(shù)顯著下降。發(fā)酵液pH7.0時(shí),菌劑活菌數(shù)最高。載體種類對(duì)菌劑質(zhì)量影響十分顯著。麩皮、硅藻土和活性炭作載體時(shí),菌劑有效活菌數(shù)相近。次面粉,玉米粉作菌劑載體效果較差。當(dāng)添加葡萄糖、淀粉、糊精時(shí),對(duì)菌劑活菌數(shù)影響不顯著。枯草芽孢桿菌菌劑最佳制備工藝:發(fā)酵液與載體比例為1.5:1、干燥溫度70℃、菌液pH為7.0、載體為麩皮,水分含量為3.3%,菌劑有效活菌數(shù)為26億/g。
[Abstract]:Rotting bacteria mainly refers to a class of microorganism agents which are studied and produced for the treatment of difficult decomposable organic matter such as animal manure, plant residues and some domestic wastes, and are widely used in crop stalks. Bacillus subtilis is a aerobic bacillus widely found in nature. It can produce enzymes such as protease, amylase, cellulase and hemicellulase. The number of spores and the number of viable bacteria are the important indexes of the quality of the rotting agent. The production cost is also an important issue concerned by the microbial agent production enterprises. The spore production conditions of Bacillus subtilis DK36 were studied. The fermentation process was optimized. On the basis of this, the preparation process of the bacterial agent was studied. In order to reduce the production cost, The DK36 of Bacillus subtilis cultured with desphenol cottonseed protein as nitrogen source was studied. The main results were as follows: 1) the spore production conditions of Bacillus subtilis DK36 were studied. When glucose, sucrose, fructose, soluble starch, corn meal, glucose, sucrose, fructose, soluble starch, corn flour, When wheat bran was carbon source, bacteria grew well. When glucose and sucrose were used as carbon source, no spores were produced. When wheat bran was used as carbon source, spore formation was the most, followed by corn flour, soluble starch, peptone, yeast extract, soybean peptone. Soybean cake powder, potassium nitrate, sodium nitrate and ammonium nitrate can make the bacteria form spores in large quantities, among which soybean peptone is the best. Urea inhibits the growth of bacteria and spore formation. Addition of Ca2 to mg _ 2, Na, Fe _ 2 and mn _ 2 can promote the formation of spores. The best medium for spores production was: bran 15.0 g / L, soybean peptone 5.0 g / L, manganese sulfate 0.1 g / L, calcium carbonate 3.0 g / L. A study on the culture of Bacillus subtilis DK36 with denatured cottonseed protein at 200rmin, inoculation at 2um, initial pH 7.0, spore number 3.6 脳 109cfumL. 2). When cottonseed protein was used as nitrogen source, the bacteria grew well, and the species of carbon source had a significant effect on spores production. Sucrose, corn flour and bran all made the bacteria grow well, but glucose and sucrose were the carbon source, the bacteria did not form spores, but corn flour and bran could make the bacteria produce a large number of spores. The addition of Ca2, mn 2, mg 2 and K could obviously promote the formation of spores. It was also found that the combination of potassium salt and magnesium salt, manganese salt, calcium salt is better. Dephenol cottonseed protein as a nitrogen source causes slow spores formation. The best medium for dephenol cottonseed protein culture of Bacillus subtilis is corn flour 30.0g / L, cottonseed protein 30.0g / L, magnesium sulfate 2.0 g / L, potassium dihydrogen phosphate 4.5 g / L. its optimum fermentation conditions are as follows:. The culture temperature is 30 鈩，

本文編號(hào)：1506585