生物表面活性剂在固态发酵中对木质纤维素生物降解的影响研究
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摘要
生物表面活性剂主要是由微生物代谢过程中分泌的具有一定生物活性的代谢产物,像合成表面活性剂一样,生物表面活性剂由亲水基和疏水基两部分组成,但生物表面活性剂比合成表面活性剂更具潜在的优势,因而随着人类环保意识的增强,更加快了生物表面活性剂取代合成表面活性剂的发展进程。本文考察了铜绿假单胞菌(Pa AB93066)产鼠李糖脂的最佳培养条件,采用了白腐菌固态发酵降解稻草的方法,选取生物表面活性剂二鼠李糖脂添加到反应中,通过分析表面活性剂对木质素过氧化物酶,锰过氧化物酶,溶解性有机碳,木质素降解率等参数的影响,考察二鼠李糖脂作用下微生物降解木质素的特性。
通过单因素试验,铜绿假单胞菌(Pa AB93066)的最佳碳源和氮源分别是花生油和NaNO3。正交试验的结果表明,铜绿假单胞菌(Pa AB93066)具有较强的生物表面活性剂生产能力且最佳培养条件为：花生油浓度125g／L,硝酸钠浓度2.5g／L,温度为34℃,摇床转速为90r／min,pH为7。在此条件下培养了180h,鼠李糖脂产量最终为16.29g／L。在最佳培养发酵条件下,随着糖脂的生产,表面张力值迅速降低,最低达到29.19mN／m,最后趋于平衡,通过对培养液进行稀释,培养液表现出临界胶束的性质。说明产生了表面活性剂且表面活性稳定。鼠李糖脂产量随菌体量的增加而增加,在菌体培养的第84h到第144h之间,鼠李糖脂产量最高,以后菌体量开始下降,鼠李糖脂产量也相应减少。所以确定提取鼠李糖脂的最佳时间为菌体生长的稳定后期到衰减初期之间,即108h。
通过固态发酵方式采用白腐菌对稻草中木质纤维素进行降解,并研究了生物表面活性剂二鼠李糖脂对该降解过程的影响.结果表明,不同浓度的二鼠李糖脂能在不同程度上提高降解过程水溶性有机碳的含量,添加0.007%和0.021%二鼠李糖脂的实验组最高TOC浓度较对照组分别提高了83.6%和54.5%,这有利于黄孢原毛平革菌的生长,且延缓了菌体的衰退.添加0.007%和0.021%浓度的二鼠李糖脂可使LiP酶活分别提高85.7%和41.2%,二鼠李糖脂对MnP酶活没有显著影响.生物表面活性剂的介入促进了白腐菌对稻草中木质素的降解,添加0.007%二鼠李糖脂可使木质素降解率提高54%.
Biosurfactants mainly are producted by microbial metabolism in the secretion of biologically active metabolites, the same as synthetic surfactants, biosurfactants by the hydrophilic and hydrophobic group of two parts, but the ratio of synthesis of biosurfactant more surface active agents of potential advantages, so as human awareness of environmental protection, but also accelerated the biosurfactant replace Synthesis of Surface-active agent development process. This article examines the Pseudomonas aeruginosa (Pa AB93066) rhamnolipid produced the best culture conditions, using the white rot fungi degrade straw solid-state fermentation method, select the biosurfactant rhamnolipidⅡadded to the reaction, Through analysis of surfactants on the lignin peroxidase, manganese peroxidase, dissolved organic carbon, lignin degradation rate parameters were investigated under the action of two rhamnolipid characteristics of microbial degradation of lignin.
Single factor test, Pseudomonas aeruginosa (Pa AB93066) the best carbon and nitrogen are peanut oil and NaNO3. Orthogonal experimental results show that Pseudomonas aeruginosa (Pa AB93066) has a strong biosurfactant production and optimal culture conditions are:peanut oil concentration 125g/L, sodium nitrate concentration 2.5g/L, temperature 34℃, shaking the 90r/min, pH is 7. Under this condition, trained 180h, rhamnolipid production ultimately 16.29g/L. Optimum culture conditions in fermentation, with the production of glycolipids, surface tension decreased rapidly, the minimum to 29.19mN/m, finally becomes more balanced, by dilution of the culture medium, medium to show the nature of the critical micelle. Showing that the surfactant and surfactant stability. Rhamnolipid production with the increase of biomass in cell culture between the first 84h to 144h, the highest rhamnolipid yield, after the cell began to decrease, rhamnolipid production is also reduced. Therefore, extraction of rhamnolipid to determine the best time for the stable growth of bacteria between late early decay,108h.
Effects of dirhamnolipid on Solid state fermentation of straw lignocellulose by white rot fungi was studied for 32 days. The results showed that dirhamnolipid at different concentrations had certain stimulative effects on total organic carbon (TOC), dirhamnolipid at 0.007%and 0.021%increased TOC by 83.6%and 54.5%, what is progressive for growth of Phanerochaetechrysosporium, and put off fungal decline. Dirhamnolipid at different concentrations had certain stimulative effects on LiP activity, the Critical Micelle Concentration 0.007%increased 85.7%, and dirhamnolipid at 0.021%increased just 41.2%. Dirhamnolipid have no stimulative effects on MnP activity. The degradation rate of lignin with rhamnolipid at 0.007%was higher than that of the control, increased 54%.
通过单因素试验,铜绿假单胞菌(Pa AB93066)的最佳碳源和氮源分别是花生油和NaNO3。正交试验的结果表明,铜绿假单胞菌(Pa AB93066)具有较强的生物表面活性剂生产能力且最佳培养条件为：花生油浓度125g／L,硝酸钠浓度2.5g／L,温度为34℃,摇床转速为90r／min,pH为7。在此条件下培养了180h,鼠李糖脂产量最终为16.29g／L。在最佳培养发酵条件下,随着糖脂的生产,表面张力值迅速降低,最低达到29.19mN／m,最后趋于平衡,通过对培养液进行稀释,培养液表现出临界胶束的性质。说明产生了表面活性剂且表面活性稳定。鼠李糖脂产量随菌体量的增加而增加,在菌体培养的第84h到第144h之间,鼠李糖脂产量最高,以后菌体量开始下降,鼠李糖脂产量也相应减少。所以确定提取鼠李糖脂的最佳时间为菌体生长的稳定后期到衰减初期之间,即108h。
通过固态发酵方式采用白腐菌对稻草中木质纤维素进行降解,并研究了生物表面活性剂二鼠李糖脂对该降解过程的影响.结果表明,不同浓度的二鼠李糖脂能在不同程度上提高降解过程水溶性有机碳的含量,添加0.007%和0.021%二鼠李糖脂的实验组最高TOC浓度较对照组分别提高了83.6%和54.5%,这有利于黄孢原毛平革菌的生长,且延缓了菌体的衰退.添加0.007%和0.021%浓度的二鼠李糖脂可使LiP酶活分别提高85.7%和41.2%,二鼠李糖脂对MnP酶活没有显著影响.生物表面活性剂的介入促进了白腐菌对稻草中木质素的降解,添加0.007%二鼠李糖脂可使木质素降解率提高54%.
Biosurfactants mainly are producted by microbial metabolism in the secretion of biologically active metabolites, the same as synthetic surfactants, biosurfactants by the hydrophilic and hydrophobic group of two parts, but the ratio of synthesis of biosurfactant more surface active agents of potential advantages, so as human awareness of environmental protection, but also accelerated the biosurfactant replace Synthesis of Surface-active agent development process. This article examines the Pseudomonas aeruginosa (Pa AB93066) rhamnolipid produced the best culture conditions, using the white rot fungi degrade straw solid-state fermentation method, select the biosurfactant rhamnolipidⅡadded to the reaction, Through analysis of surfactants on the lignin peroxidase, manganese peroxidase, dissolved organic carbon, lignin degradation rate parameters were investigated under the action of two rhamnolipid characteristics of microbial degradation of lignin.
Single factor test, Pseudomonas aeruginosa (Pa AB93066) the best carbon and nitrogen are peanut oil and NaNO3. Orthogonal experimental results show that Pseudomonas aeruginosa (Pa AB93066) has a strong biosurfactant production and optimal culture conditions are:peanut oil concentration 125g/L, sodium nitrate concentration 2.5g/L, temperature 34℃, shaking the 90r/min, pH is 7. Under this condition, trained 180h, rhamnolipid production ultimately 16.29g/L. Optimum culture conditions in fermentation, with the production of glycolipids, surface tension decreased rapidly, the minimum to 29.19mN/m, finally becomes more balanced, by dilution of the culture medium, medium to show the nature of the critical micelle. Showing that the surfactant and surfactant stability. Rhamnolipid production with the increase of biomass in cell culture between the first 84h to 144h, the highest rhamnolipid yield, after the cell began to decrease, rhamnolipid production is also reduced. Therefore, extraction of rhamnolipid to determine the best time for the stable growth of bacteria between late early decay,108h.
Effects of dirhamnolipid on Solid state fermentation of straw lignocellulose by white rot fungi was studied for 32 days. The results showed that dirhamnolipid at different concentrations had certain stimulative effects on total organic carbon (TOC), dirhamnolipid at 0.007%and 0.021%increased TOC by 83.6%and 54.5%, what is progressive for growth of Phanerochaetechrysosporium, and put off fungal decline. Dirhamnolipid at different concentrations had certain stimulative effects on LiP activity, the Critical Micelle Concentration 0.007%increased 85.7%, and dirhamnolipid at 0.021%increased just 41.2%. Dirhamnolipid have no stimulative effects on MnP activity. The degradation rate of lignin with rhamnolipid at 0.007%was higher than that of the control, increased 54%.
引文
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