【作者】 林元山；

【导师】 梁智群；

【作者基本信息】 广西大学 ， 发酵工程， 2004， 硕士

【摘要】 糖蜜酒精废液是糖厂的主要污染源之一，是一种深褐色、高酸性、高浓度有机废水。广西大学食品与发酵工程所研究其综合治理时，发现糖蜜酒精废液有促进康氏木霉产纤维素酶的作用，本论文为此作了深入研究。 论文研究了糖蜜酒精废液诱导下的纤维素酶酶学特性：FP，EG，BG，CBH酶的最适pH分别为4.8，4.8，4.8，5.0；最适作用温度分别为50℃，70℃，60℃，55℃；测定了CBH，EG，BG酶的米氏常数；探讨了底物浓度、稀释倍数、金属离子等因素对FP，EG，BG，CBH酶的影响。 论文分离纯化了纤维素酶：EG，BG酶经DEAE C52，CL-6B柱层析分离，酶活力最终回收率分别为2.06％，65.11％，纯化倍数分别为24.55、169.84，比活力分别达189.48IU／mg蛋白，656.28IU／mg蛋白。CBH酶经DEAE-C52分离，回收率和纯化倍数分别为85.61％，31.56。 论文研究分析了糖蜜酒精废液的诱导机理：表明糖蜜酒精废液对纤维素酶的产生具有诱导效应，而非营养因素的作用；其FP酶活力是Mandels营养液的4.8倍；EG，BG，CBH酶活力是Mandels营养液的3～5倍。经电泳测定，糖蜜酒精废液诱导康氏木霉大量表达胞外蛋白p^17.8，p^72.9；胞外蛋白p^17.8，p^72.9不具有EG，BG，CBH酶活性，即无纤维素酶活性。更多还原

【Abstract】 The waste molasses of ethanol, deep brow, high acid, condensed waste organic water, is one of the main puollution sourses from sugar refineries. During the comprehensive disposal of the waste performed by Food and Fermentation engineering Institute of Guangxi University, the waste molasses of ethanol was found to be helpful to the excretion of cellulase from Trichoderma Knoningii. The further studies on cellulase from Trichoderma Knoningii affected by the waste are achieved in this thesis.Firstly, the enzymic characteristics of cellulase from waste molasses medium, including FP, EG, BG and CBH are determined in this thesis. The optimum pH for catalyzed reactions of FP, EG, BG and CBH are 4.8, 4.8, 4.8, 5.0, respectively. The optimum temperature for catalyzed reactions of FP, EG, BG and CBH are 50℃, 70℃, 60℃, 55℃, respectively. The michaelis constant of EG, BG and CBH, as well as the effects of substrates, dilutions and ions on FP, EG, BG and CBH are assayed.Secondly, purification and isolation of cellulase are performed in the thesis. By means of ammonium sulphate precipitation, anion exchange chromatography column DEAE cellulose 52 and DEAE sepharose CL-6B, the recoveries of EG and BG reach 2.06% and 65.11%, purity of EG and BG reach 24.55 times and 169.84 times, specfic activities of EG and BG reach189.48IU per mg protein and 656.28IU per mg protein, respectively.Thirdly, the mechanism of cellulase induced by waste molasses of ethanol from Trichoderma Knoningii is investigated. It’s the function of inducement, not function of nourishment that cellulase is excreted excessively from Trichoderma Knoningii on the medium of waste molasses. The activity of FP is 4.8 times as much as that of Mandels medium, comparative with EG, BG and CBH are 3 to 5 times as much as that of Mandels medium. Analyzed by SDS-PAGE, waste molasses of ethanol can induce two proteins over-Expression from Trichoderma Knoningi. These proteins, named P17.8andP72.9 according to molecular weight 17.8KD, 72.9KD respectively, have no activities of EG, BG and CBH. So P17.8and P72.9 have not activity of cellulase. They are not cellulase.更多还原