【正文】 本科畢業(yè)論文（設(shè)計(jì)）論文（設(shè)計(jì)）題目：降解酒糟生物質(zhì)的纖維素分解細(xì)菌的篩選及產(chǎn)酶研究學(xué) 院： 農(nóng) 學(xué) 院 ＿專 業(yè)： 農(nóng)業(yè)資源與環(huán)境 班 級：＿09 農(nóng) 資＿ ＿學(xué) 號：＿0909010165 ＿ 學(xué)生姓名：＿ ＿ ＿＿指導(dǎo)教師：＿ ＿2013年 5 月 28 日貴州大學(xué)本科畢業(yè)論文（設(shè)計(jì)）誠信責(zé)任書本人鄭重聲明：本人所呈交的畢業(yè)論文（設(shè)計(jì)），是在導(dǎo)師的指導(dǎo)下獨(dú)立進(jìn)行研究所完成。畢業(yè)論文（設(shè)計(jì)）中凡引用他人已經(jīng)發(fā)表或未發(fā)表的成果、數(shù)據(jù)、觀點(diǎn)等，均已明確注明出處。特此聲明。論文（設(shè)計(jì)）作者簽名： 日 期： IV 貴 貴州大學(xué)本科畢業(yè)論文（設(shè)計(jì)） 目 錄摘 要 IIAbstract III1前言 1 1 1 22材料與方法 3 3 3 4 4 4 5 5 5 6 純化 7 7 7 83結(jié)果與分析 純化結(jié)果 10 10 10 12 12 13 154結(jié)果與討論 17參考文獻(xiàn) 19致 謝 20降解酒糟生物質(zhì)的纖維素分解細(xì)菌的篩選及產(chǎn)酶研究摘 要本試驗(yàn)從茅臺酒廠提供的樣品中分別從30℃和50℃下分離出能分解利用羧甲基纖維素（CMCNa）的細(xì)菌共244株。經(jīng)革蘭氏碘液染色，，其中，菌株BL595的透明圈與菌落半徑比值最大。在這103株菌中， cm（30℃） cm（50℃）的菌株有20株。本試驗(yàn)分別從50176。C和30℃培養(yǎng)篩選條件下選取7株和6株，對這些菌株進(jìn)行纖維素酶活及其他各項(xiàng)指標(biāo)的測定。菌株的生長曲線試驗(yàn)分別在30℃（中溫）條件下靜置培養(yǎng)和50℃（高溫）條件下振蕩進(jìn)行培養(yǎng)。通過不同溫度分離篩選條件下得到的菌株分別在其對應(yīng)溫度下的生長曲線可以看出，高溫振蕩培養(yǎng)的菌株會在試管表面長出菌苔，菌液澄清，試管底部有沉淀；中溫靜置培養(yǎng)條件下的菌株菌液整體渾濁，分布均勻。高溫振蕩培養(yǎng)的菌株在培養(yǎng)前期（12 h）生長快于低溫靜置培養(yǎng)的菌株，但培養(yǎng)3 d后，二者生長趨勢趨向一致，但中溫條件下篩選的部分菌株（BL5106）性能優(yōu)于高溫條件下篩選得到的菌株。通過纖維素酶活力鑒定，菌株BL5106的酶活力最高， U/mL，表明該菌株最具研究價(jià)值，該探究也為下一階段的堆肥試驗(yàn)提供了前期準(zhǔn)備及研究用菌。關(guān)鍵詞：纖維素分解細(xì)菌，酶活性，纖維素酶，篩選Screening of Vinasse Cellulose Deposing Bacteria and Research on Enzyme ProductionAbstract244 bacteria strains which could form a sharp and distinct zone around the cellulaseproducing microbial colonies on carboxymethylcellulose (CMC) plates with Gram’s iodine had been isolated. There were 103 strains which the radius ratio of distinct zone and colony was more than , and among which, the radius ratio was more than were 12 strains. Especially, the radius ratio of strain BL595 was the highest (). The strain number which the radius of distinct zone was more than cm under 30℃ and more than cm under 50℃ conditions was 20. In this study, 7 strains from 50℃ conditions and 6 strains from 30℃ conditions had been chosen to assay the active of cellulases and other index.The growth curve of strains test were cultured under 30℃ static culture and 50℃ oscillation conditions, respectively. There were lawns appearing on the culture medium surface, and the liquid were clarification under oscillation conditions. However, there was precipitation in the bottom of the test tube. In contrast, the liquid were overall turbidity under static conditions. Strains cultured under 50℃ oscillation conditions grew faster than the strains from 30℃ conditions during the earlier incubation stage, however, the growth from different conditions were consistency after incubation 3 d. The performance of strain BL516 from 30℃ conditions was better than the strains from 50℃ conditions.The cellulose active of BL5106 was the highest, which was U/mL. This study provides preparation and microbes for the posting experiment.Keywords: cellulose deposing bacteria, enzymatic activity, cellulase activity, screening第 1 頁 貴 貴州大學(xué)本科畢業(yè)論文（設(shè)計(jì)） 1前言酒糟是釀酒業(yè)最主要的副產(chǎn)物，在我國，每年白酒酒糟產(chǎn)量都在2500萬t以上[1]。由于酒糟酸度偏大，可生化性較較強(qiáng)，極易腐爛，不便存儲，若不及時(shí)處理，易對環(huán)境造成極大的污染[2]。酒糟生物質(zhì)主要由纖維素和木質(zhì)素構(gòu)成，同時(shí)含有一定的殘余糖分、淀粉、粗脂肪和蛋白質(zhì)[3]，因此，可利用具有降解纖維素的菌種對其進(jìn)行分解，把其中部分纖維素轉(zhuǎn)化為有機(jī)質(zhì)等，從而提高酒糟的綜合利用效率[4]。對纖維素酶產(chǎn)生菌的研究過去多集中于真菌的研究和應(yīng)用，鑒于細(xì)菌繁殖和產(chǎn)酶速度快、發(fā)酵周期短，有利于工業(yè)生產(chǎn)，故近年來對細(xì)菌產(chǎn)纖維素酶日益引起重視。纖維素酶的比活力一般都很低，因而產(chǎn)酶成本高。據(jù)估計(jì)，纖維素水解成葡萄糖所需的酶蛋白要比淀粉相應(yīng)水解所需的酶蛋白多100倍，這是影響纖維素酶實(shí)際應(yīng)用的重要原因之一[5]。因而，篩選纖維素酶比活力高的菌株無疑具有重要的意義。本試驗(yàn)針對酒糟生物質(zhì)降解慢的難題，擬從茅臺酒廠提供的樣品中分離細(xì)菌，篩選出酶活性較高的纖維素細(xì)菌菌株，對篩選的菌株進(jìn)行產(chǎn)酶特性初步研究，以此為依據(jù)優(yōu)化產(chǎn)酶條件，以期為酒糟綜合利用研究提供科學(xué)依據(jù)。能產(chǎn)纖維素酶的細(xì)菌種類雖不如真菌，生孢食纖維菌屬( Sporocyt