以木质纤维素为原料的燃料乙醇生产工艺及废水零排放技术研究
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摘要
在环境污染和能源危机的背景下,燃料乙醇作为一种清洁可再生的能源得到了众多的关注。木质纤维素原料来源广泛,以其为原料生产燃料乙醇可以为避免粮食危机作出贡献。但目前木质纤维素生产燃料乙醇存在工艺复杂,费用居高不下,环境污染等问题。本课题选择玉米秸秆等农林废料为基本原料,研究了燃料乙醇的制备工艺,分析了电催化法木质纤维素水解液中发酵抑制物的控制及分离方法,构建了固定化混合菌种发酵工艺及母液循环利用的方法,设计了一套通过乙醇废水生物絮凝处理和循环利用来实现乙醇废水零排放的技术方案。
探究了木质纤维素水解液中水解液抑制物对驯化前后的酵母菌生长和乙醇发酵的影响。实验结果表明,乙酸、糠醛、甲酸、苯酚都会对酵母菌的生长繁殖产生抑制作用,四种物质对酵母菌生长的抑制强度依次为甲酸>糠醛>苯酚>乙酸,四种物质对酒精发酵的抑制强度依次为甲酸>糠醛>苯酚>乙酸,四种抑制物对未驯化的酵母发酵的抑制作用比对驯化后的强。
探究了木质纤维素水解液中的抑制物对木糖发酵的影响,通过对比在不同浓度的抑制物作用下的嗜单宁管囊酵母菌的生长曲线,以及菌种发酵木糖过程所消耗的糖和乙醇产率来概括了木质纤维素水解液中抑制物对发酵的影响程度,实验证明抑制物可以减缓或者完全抑制菌种的生长和木糖的发酵过程。且驯化后的菌种表现出更好的耐毒性。
考察了水解液的脱毒方法,确定了脱毒工艺。调节水解液pH至6,CTS投量0.1g/L,搅拌均匀,静置过滤。所获的最佳的脱毒率为78.5%。基于响应面试验设计所得的脱毒工艺的最佳工艺参数准确可靠,且具有实用价值。且在该方法处理后的水解液中菌种能很快地进入生长平衡期。
为了充分利用木质纤维素水解液中的葡萄糖和木糖发酵,采用固定化混合菌种(同时固定驯化后的嗜单宁管囊酵母和驯化后的酿酒酵母两种菌种)进行絮凝脱毒后水解液的发酵,同时对普通的固定化颗粒进行改性,研究表明壳聚糖-海藻酸钙固定化菌种有较好的发酵效果,且此时氯化钙的浓度为3%,固定化粒子的粒径为5.3 mm。此种固定化混合菌种用于发酵,发酵温度30℃,震荡培养器转速150 r/min,木质纤维素水解液pH5.03,发酵液还原糖浓度82g/L时发酵效果最佳,酒精浓度达20.04g／L。且各种菌种最佳发酵条件下的乙醇产率排序为：海藻酸钙-壳聚糖固定化混合菌种>海藻酸钙-三氧化二铝固定化混合菌种>游离态的混合菌种>海藻酸钙-壳聚糖固定化酿酒酵母>海藻酸钙-壳聚糖固定化嗜单宁管囊酵母>海藻酸钙-三氧化二铝固定化酿酒酵母>海藻酸钙-三氧化二铝固定化嗜单宁管囊酵母>游离态的酿酒酵母>游离态的嗜单宁管囊酵母。
本工作还提出和设计了一套乙醇废水生物絮凝处理和循环利用技术方案,实现废水零排放。试验结果表明,以本实验室合成的聚天冬氨酸为生物絮凝剂,可有效分离废水中的杂质,消除影响乙醇发酵的有害因素,废水经过十次循环处理利用,乙醇收率仍与使用自来水相当,达到应用标准。
探索性研究循环催化法处理木质纤维素原料。具体的工艺过程为将原料与70%的浓硫酸混合,反应温度80℃,形成胶状混合物后将酸浓度稀释为35%,沸腾反应一小时后过滤,使用强酸性树脂分离。分离的最终结果显示树脂分离糖酸有一定的可行性。
Environmental pollution and energy crisis have brought a large number of concerns on the research of fuel ethanol as a kind of clean and renewable energy. Wide range of lignocellulosic materials as raw materials for the production of fuel ethanol can effectively avoid the food crisis. But the process was complex, with high cost and environmental pollution. Forestry wastes such as corn straw was selected as the basic raw material for the ethanol production research, and the fermentation inhibition of the hydrolyzate from electro-catalytic hydrolysis treatment was controlled and separated. The mixed yeasts immobilized fermentation process was designed and biological flocculation wastewater treatment were constructed in order to realize wastewater zero-discharge.
The inhibitory effects of the hazardous materials in lignocellulose acid-hydrolysates on glucose fermentation and yeast growth were studied. The results showed that acetic acid, furfural, formic acid, and phenol had the inhibition of growth and reproduction of yeast, and the four substances on the yeast growth inhibition strength was in the order of formic acid> furfural> phenol> acetic acid, at the same time, four substances inhibition on alcoholic fermentation was followed by formic acid> furfural> phenol> acetic acid. The yeast after domestication showed better stability.
The inhibitory effects of the hazardous materials in lignocellulose acid-hydrolysates on xylose fermentation were studied. Through the growth curve of yeast and the fermentation rate, the inhibitions of the hazardous materials were measured. The result showed that these inhibitions could make the yeast stop their growth and fermentation. And the strain after domestication showed stronger anti-inhibitor capability than unacclimated strain.
The detoxification of acid hydrolyzate was studied in this paper. CTS was confirmed as the biological flocculant which has a good detoxification effect. The detoxification process was that the hydrolyzate after filtration was treated to get the solution with pH of 6, and then 0.1 g/L CTS was added to the solution. After reaction, the solution was filtrated and the detoxification process was finished. The best detoxification rate was 78.5%. The Box-Behnken design was accurate, reliable and with practical value. The yeast showed good adaptability with hydrolyzate after detoxification.
In order to achieve a higher ethanol fermentation capacity and faster ethanol fermentation rate, the co-immobilized fermentation was used. At the same time, the immobilized particles were modified. The result showed that the CTS-calcium alginate was the best immobilized material when the CaCl2 concentraion was 3% and particle size was 5.3 mm. The optimum fermentation conditions at 30℃and 150 rpm in shaking flasks were as follows:pH 5.03 and reducing sugar 82 g/L. The final ethanol concentration was 20.04 g/L. At the same time the fermentation of different yeast was compared. The ethanol yield of yeast fermentation under optimum conditions was in this order:CTS-calcium alginate immobilized mixed yeasts> Al2O3-calcium alginate immobilized mixed yeasts> free-state mixed yeasts> CTS-calcium alginate immobilized Saccharomyces cerevisiae> CTS-calcium alginate immobilized Pachysolen tannophilus> Al2O3-calcium alginate immobilized Saccharomyces cerevisiae> Al2O3-calcium alginate immobilized Pachysolen tannophilus> free-state Saccharomyces cerevisiae> free-state Pachysolen tannophilus.
In the present study, a wastewater treatment system for the ethanol fermentation industry was developed by recycling distillery wastewater and the wastewater zero discharge was achieved. Bio-flocculation was shown to be an effective way to diminish the content of inhibitory compounds drastically when the waste was recirculated. The recycle of wastewater without flocculation showed negative effects on ethanol yield as recycling was repeated. This new process was confirmed to have stable operation over ten recycles.
Exploratory study of lignocellulosic cyclic catalytic treatment was studied. The lignocellulose materials was treated by 70% concentrated sulfuric acid at 80℃to get a gel state and then the acid was diluted to 35% and the reaction was continued for one hour at boiling situation. The filtrate was treated by stong acid resin to get the acid and sugar separation. And the result showed the final separation of sugar and acid reached a certain extent.
探究了木质纤维素水解液中水解液抑制物对驯化前后的酵母菌生长和乙醇发酵的影响。实验结果表明,乙酸、糠醛、甲酸、苯酚都会对酵母菌的生长繁殖产生抑制作用,四种物质对酵母菌生长的抑制强度依次为甲酸>糠醛>苯酚>乙酸,四种物质对酒精发酵的抑制强度依次为甲酸>糠醛>苯酚>乙酸,四种抑制物对未驯化的酵母发酵的抑制作用比对驯化后的强。
探究了木质纤维素水解液中的抑制物对木糖发酵的影响,通过对比在不同浓度的抑制物作用下的嗜单宁管囊酵母菌的生长曲线,以及菌种发酵木糖过程所消耗的糖和乙醇产率来概括了木质纤维素水解液中抑制物对发酵的影响程度,实验证明抑制物可以减缓或者完全抑制菌种的生长和木糖的发酵过程。且驯化后的菌种表现出更好的耐毒性。
考察了水解液的脱毒方法,确定了脱毒工艺。调节水解液pH至6,CTS投量0.1g/L,搅拌均匀,静置过滤。所获的最佳的脱毒率为78.5%。基于响应面试验设计所得的脱毒工艺的最佳工艺参数准确可靠,且具有实用价值。且在该方法处理后的水解液中菌种能很快地进入生长平衡期。
为了充分利用木质纤维素水解液中的葡萄糖和木糖发酵,采用固定化混合菌种(同时固定驯化后的嗜单宁管囊酵母和驯化后的酿酒酵母两种菌种)进行絮凝脱毒后水解液的发酵,同时对普通的固定化颗粒进行改性,研究表明壳聚糖-海藻酸钙固定化菌种有较好的发酵效果,且此时氯化钙的浓度为3%,固定化粒子的粒径为5.3 mm。此种固定化混合菌种用于发酵,发酵温度30℃,震荡培养器转速150 r/min,木质纤维素水解液pH5.03,发酵液还原糖浓度82g/L时发酵效果最佳,酒精浓度达20.04g／L。且各种菌种最佳发酵条件下的乙醇产率排序为：海藻酸钙-壳聚糖固定化混合菌种>海藻酸钙-三氧化二铝固定化混合菌种>游离态的混合菌种>海藻酸钙-壳聚糖固定化酿酒酵母>海藻酸钙-壳聚糖固定化嗜单宁管囊酵母>海藻酸钙-三氧化二铝固定化酿酒酵母>海藻酸钙-三氧化二铝固定化嗜单宁管囊酵母>游离态的酿酒酵母>游离态的嗜单宁管囊酵母。
本工作还提出和设计了一套乙醇废水生物絮凝处理和循环利用技术方案,实现废水零排放。试验结果表明,以本实验室合成的聚天冬氨酸为生物絮凝剂,可有效分离废水中的杂质,消除影响乙醇发酵的有害因素,废水经过十次循环处理利用,乙醇收率仍与使用自来水相当,达到应用标准。
探索性研究循环催化法处理木质纤维素原料。具体的工艺过程为将原料与70%的浓硫酸混合,反应温度80℃,形成胶状混合物后将酸浓度稀释为35%,沸腾反应一小时后过滤,使用强酸性树脂分离。分离的最终结果显示树脂分离糖酸有一定的可行性。
Environmental pollution and energy crisis have brought a large number of concerns on the research of fuel ethanol as a kind of clean and renewable energy. Wide range of lignocellulosic materials as raw materials for the production of fuel ethanol can effectively avoid the food crisis. But the process was complex, with high cost and environmental pollution. Forestry wastes such as corn straw was selected as the basic raw material for the ethanol production research, and the fermentation inhibition of the hydrolyzate from electro-catalytic hydrolysis treatment was controlled and separated. The mixed yeasts immobilized fermentation process was designed and biological flocculation wastewater treatment were constructed in order to realize wastewater zero-discharge.
The inhibitory effects of the hazardous materials in lignocellulose acid-hydrolysates on glucose fermentation and yeast growth were studied. The results showed that acetic acid, furfural, formic acid, and phenol had the inhibition of growth and reproduction of yeast, and the four substances on the yeast growth inhibition strength was in the order of formic acid> furfural> phenol> acetic acid, at the same time, four substances inhibition on alcoholic fermentation was followed by formic acid> furfural> phenol> acetic acid. The yeast after domestication showed better stability.
The inhibitory effects of the hazardous materials in lignocellulose acid-hydrolysates on xylose fermentation were studied. Through the growth curve of yeast and the fermentation rate, the inhibitions of the hazardous materials were measured. The result showed that these inhibitions could make the yeast stop their growth and fermentation. And the strain after domestication showed stronger anti-inhibitor capability than unacclimated strain.
The detoxification of acid hydrolyzate was studied in this paper. CTS was confirmed as the biological flocculant which has a good detoxification effect. The detoxification process was that the hydrolyzate after filtration was treated to get the solution with pH of 6, and then 0.1 g/L CTS was added to the solution. After reaction, the solution was filtrated and the detoxification process was finished. The best detoxification rate was 78.5%. The Box-Behnken design was accurate, reliable and with practical value. The yeast showed good adaptability with hydrolyzate after detoxification.
In order to achieve a higher ethanol fermentation capacity and faster ethanol fermentation rate, the co-immobilized fermentation was used. At the same time, the immobilized particles were modified. The result showed that the CTS-calcium alginate was the best immobilized material when the CaCl2 concentraion was 3% and particle size was 5.3 mm. The optimum fermentation conditions at 30℃and 150 rpm in shaking flasks were as follows:pH 5.03 and reducing sugar 82 g/L. The final ethanol concentration was 20.04 g/L. At the same time the fermentation of different yeast was compared. The ethanol yield of yeast fermentation under optimum conditions was in this order:CTS-calcium alginate immobilized mixed yeasts> Al2O3-calcium alginate immobilized mixed yeasts> free-state mixed yeasts> CTS-calcium alginate immobilized Saccharomyces cerevisiae> CTS-calcium alginate immobilized Pachysolen tannophilus> Al2O3-calcium alginate immobilized Saccharomyces cerevisiae> Al2O3-calcium alginate immobilized Pachysolen tannophilus> free-state Saccharomyces cerevisiae> free-state Pachysolen tannophilus.
In the present study, a wastewater treatment system for the ethanol fermentation industry was developed by recycling distillery wastewater and the wastewater zero discharge was achieved. Bio-flocculation was shown to be an effective way to diminish the content of inhibitory compounds drastically when the waste was recirculated. The recycle of wastewater without flocculation showed negative effects on ethanol yield as recycling was repeated. This new process was confirmed to have stable operation over ten recycles.
Exploratory study of lignocellulosic cyclic catalytic treatment was studied. The lignocellulose materials was treated by 70% concentrated sulfuric acid at 80℃to get a gel state and then the acid was diluted to 35% and the reaction was continued for one hour at boiling situation. The filtrate was treated by stong acid resin to get the acid and sugar separation. And the result showed the final separation of sugar and acid reached a certain extent.
引文
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