纤维素热化学催化液化反应及微生物发酵综合利用
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摘要
纤维素是葡萄糖分子以β-1，4糖苷键结合而成的高分子化合物，一般聚合度为3,500-10,000。纤维素是植物细胞壁的主要成分，广泛存在于自然界，是地球上最丰富、最廉价的可再生资源，也是人类未来能源、食品和化工原料的主要来源。随着世界人口的激增，粮食和能源的短缺将日趋严重，可再生纤维素资源的开发利用已引起全世界的普遍关注。
为了提高纤维类物质的综合利用效率，降低对环境的污染，维护生态平衡。对纤维类废弃物在浓硫酸／苯酚为催化剂、乙二醇为反应介质的热化学催化液化反应中，对其各反应物的用量比例、反应条件进行优化。并且对最优条件下的液化产物进行了初步的组分测定。实验结果表明，纤维类废弃物在浓硫酸／苯酚(浓硫酸占所加物质总量的质量分数为6％)的混合催化体系中，当温度为160℃，时间为70min时的液化效果最好。经凝胶色谱法测定表明，以浓硫酸／苯酚为催化剂、乙二醇为反应介质的反应体系，所得到液化产物的平均相对分子质量最低，M_w为806，M_n为799。经红外光谱(IR)检测结果表明，液化产物的游离羟基明显增多，其它的游离端基也是明显增加，这说明液化反应破坏了纤维素的晶格结构，而且部分水解了糖苷键。
从堆肥、垃圾填埋场堆腐物和土壤中分离并经初筛、复筛得到分解纤维素能力较强的菌共7株，外加实验室已保藏的菌5株，共计12株菌株。分别对其进行了滤纸分解度、羧甲基纤维素(CMC)酶活和天然纤维素酶活的测定。筛选出对天然纤维素分解能力较强的菌株共5株，酶活力在60.00mg／mL·d以上。通过改变其培养基中天然纤维素的含量，发现随着培养基中纤维素含量的减少，酶活力也随之降低。
进而利用这些高酶活菌株对液化产物进行利用研究。利用气质联用仪和红外光谱仪对液化产物进行了分析。结果表明，液化产物中出现甲基和亚甲基等基团的振动，并存在麦草纤维素的单体葡萄糖的衍生物，说明液化反应破坏了纤维类废弃物的晶格结构。从而打破了生物利用的禁锢，容易被微生物利用；不同菌株对于液化产物的利用方式不同，利用液化产物进行混合菌株发酵培养，真蛋白含量可达到30.74％；混合菌株利用液化产物的酒精发酵，酒精含量可达到19.0％(V／V)。
纤维类废弃物的热化学液化反应过程中，产生了一类低沸点的气态混合物。经GC-MS检测，结果显示气态混合物为一类有机酸的混合物和苯酚-硫酸形成的化合物。气态混合物作为一种微生物培养基的添加剂，用来培养球形红假单胞菌(Rhodopseudomonas sphaeroides)和沼泽红假单胞菌(Rhodopseudomonas palustris)。对细胞浓度(OD_(660nm)值)和气态混合物的残留成分测定，结果表明添加了气态混合物的光合细菌浓度明显高于未添加的，而且培养基中的气态混合物经培养一周后明显减少。实验结果表明，气态混合物能够促进光合细菌的生长，菌体蛋白含量明显增加。
Cellulose is macromolecular compound made by more glucose molecules linked through -1,4 indican bond, including 3,500-10,000. It, one of chief components of plant cell wall, lies in the earth abroad. However, cellulose is a most important recycled resources, more abundant and cheaper. With the population increasingly, it is becoming more and more indispensable to energy sources, foodstuff and raw material of industry in future. It's time to be more attention to cellulose.
In order to enhance the efficiency of usage of lignocellulosic wastes and protect the balance of ecosystem, the liquefaction of lignocellulosic wastes was studied in this paper. Lignocellulosic wastes and other chemicals were put into the reaction container and then liquefied. The effects of catalysts, solvents, reaction time and temperature on the liquefaction were investigated. The molecular weight distributions of liquefaction products were determined by GPC. In most cases, Low residue content cannot be achieved by single catalyst. However the binary catalysts system favors low residue content. It was found that the residue content were remained at a low level with ethylene glycol as the reaction media and 6%(by mass)of sulfuric acid as the catalyst in the presence of phenol. The contents of lipophilic products were investigated. The minimum residue content was found to be 8.8% and the MW and M of liquefaction products were 806 and 799 respectively. As a result, 1 ipophilic products have the highest yield when the reaction time was 70min. The identification of IR spectrometry showed that there are more dissociated hydroxy (HO-) and end-groups in the liquefaction products than in the plant cellulose. As a result, the crystal structure was destroyed in the thermochemical liquefaction.
There are twelve bacteria, conserved five in our laboratory and screened seven through dunghill, putrid garbage or soil. 1 examined decomposition rate of filter paper, CMC enzyme and crude cellulose enzyme of bacteria. Five bacteria, 60.00mg/mL d upwards, were selected. The results show that enzyme is fading away with cellulose content decreasing in the culture media.
The liquefaction products were analyzed by applying any analysis instruments including Infrared spectrum (IR) and GC-MS. The results showed that the structure of wheat straw was destroyed in liquefaction, with producing some ramification of glucose Some screened strains can decompose and use the liquefaction products. As a result of destroying crystal of lignocellulosic wastes in the liquefaction, the screened strains can produce pure protein by using liquefaction products, and its yield is 30.74%; the mixed strains can turn the liquefaction products into alcohol, and its highest content is 19.0%(V/V).
In the liquefaction of lignocellulosic wastes, there produced gaseous substances as a byproduct. By using GC-MS, the result showed that the gaseous substances are the mixture of organic acids and a compound as a byproduct in the reaction of vitriol oil and phenol. We cultured photosynthesis bacteria (PSB), using trfe mixed substances as an additive of culture medium. Through the analysis of residue of the mixed substances in the culture media and OD660nm of cell concentration, it revealed that cell concentration became higher, and residue content of the mixed substances was lower in a week. We can draw a conclusion that the mixture can promote PSB reproduction and protein content.
为了提高纤维类物质的综合利用效率，降低对环境的污染，维护生态平衡。对纤维类废弃物在浓硫酸／苯酚为催化剂、乙二醇为反应介质的热化学催化液化反应中，对其各反应物的用量比例、反应条件进行优化。并且对最优条件下的液化产物进行了初步的组分测定。实验结果表明，纤维类废弃物在浓硫酸／苯酚(浓硫酸占所加物质总量的质量分数为6％)的混合催化体系中，当温度为160℃，时间为70min时的液化效果最好。经凝胶色谱法测定表明，以浓硫酸／苯酚为催化剂、乙二醇为反应介质的反应体系，所得到液化产物的平均相对分子质量最低，M_w为806，M_n为799。经红外光谱(IR)检测结果表明，液化产物的游离羟基明显增多，其它的游离端基也是明显增加，这说明液化反应破坏了纤维素的晶格结构，而且部分水解了糖苷键。
从堆肥、垃圾填埋场堆腐物和土壤中分离并经初筛、复筛得到分解纤维素能力较强的菌共7株，外加实验室已保藏的菌5株，共计12株菌株。分别对其进行了滤纸分解度、羧甲基纤维素(CMC)酶活和天然纤维素酶活的测定。筛选出对天然纤维素分解能力较强的菌株共5株，酶活力在60.00mg／mL·d以上。通过改变其培养基中天然纤维素的含量，发现随着培养基中纤维素含量的减少，酶活力也随之降低。
进而利用这些高酶活菌株对液化产物进行利用研究。利用气质联用仪和红外光谱仪对液化产物进行了分析。结果表明，液化产物中出现甲基和亚甲基等基团的振动，并存在麦草纤维素的单体葡萄糖的衍生物，说明液化反应破坏了纤维类废弃物的晶格结构。从而打破了生物利用的禁锢，容易被微生物利用；不同菌株对于液化产物的利用方式不同，利用液化产物进行混合菌株发酵培养，真蛋白含量可达到30.74％；混合菌株利用液化产物的酒精发酵，酒精含量可达到19.0％(V／V)。
纤维类废弃物的热化学液化反应过程中，产生了一类低沸点的气态混合物。经GC-MS检测，结果显示气态混合物为一类有机酸的混合物和苯酚-硫酸形成的化合物。气态混合物作为一种微生物培养基的添加剂，用来培养球形红假单胞菌(Rhodopseudomonas sphaeroides)和沼泽红假单胞菌(Rhodopseudomonas palustris)。对细胞浓度(OD_(660nm)值)和气态混合物的残留成分测定，结果表明添加了气态混合物的光合细菌浓度明显高于未添加的，而且培养基中的气态混合物经培养一周后明显减少。实验结果表明，气态混合物能够促进光合细菌的生长，菌体蛋白含量明显增加。
Cellulose is macromolecular compound made by more glucose molecules linked through -1,4 indican bond, including 3,500-10,000. It, one of chief components of plant cell wall, lies in the earth abroad. However, cellulose is a most important recycled resources, more abundant and cheaper. With the population increasingly, it is becoming more and more indispensable to energy sources, foodstuff and raw material of industry in future. It's time to be more attention to cellulose.
In order to enhance the efficiency of usage of lignocellulosic wastes and protect the balance of ecosystem, the liquefaction of lignocellulosic wastes was studied in this paper. Lignocellulosic wastes and other chemicals were put into the reaction container and then liquefied. The effects of catalysts, solvents, reaction time and temperature on the liquefaction were investigated. The molecular weight distributions of liquefaction products were determined by GPC. In most cases, Low residue content cannot be achieved by single catalyst. However the binary catalysts system favors low residue content. It was found that the residue content were remained at a low level with ethylene glycol as the reaction media and 6%(by mass)of sulfuric acid as the catalyst in the presence of phenol. The contents of lipophilic products were investigated. The minimum residue content was found to be 8.8% and the MW and M of liquefaction products were 806 and 799 respectively. As a result, 1 ipophilic products have the highest yield when the reaction time was 70min. The identification of IR spectrometry showed that there are more dissociated hydroxy (HO-) and end-groups in the liquefaction products than in the plant cellulose. As a result, the crystal structure was destroyed in the thermochemical liquefaction.
There are twelve bacteria, conserved five in our laboratory and screened seven through dunghill, putrid garbage or soil. 1 examined decomposition rate of filter paper, CMC enzyme and crude cellulose enzyme of bacteria. Five bacteria, 60.00mg/mL d upwards, were selected. The results show that enzyme is fading away with cellulose content decreasing in the culture media.
The liquefaction products were analyzed by applying any analysis instruments including Infrared spectrum (IR) and GC-MS. The results showed that the structure of wheat straw was destroyed in liquefaction, with producing some ramification of glucose Some screened strains can decompose and use the liquefaction products. As a result of destroying crystal of lignocellulosic wastes in the liquefaction, the screened strains can produce pure protein by using liquefaction products, and its yield is 30.74%; the mixed strains can turn the liquefaction products into alcohol, and its highest content is 19.0%(V/V).
In the liquefaction of lignocellulosic wastes, there produced gaseous substances as a byproduct. By using GC-MS, the result showed that the gaseous substances are the mixture of organic acids and a compound as a byproduct in the reaction of vitriol oil and phenol. We cultured photosynthesis bacteria (PSB), using trfe mixed substances as an additive of culture medium. Through the analysis of residue of the mixed substances in the culture media and OD660nm of cell concentration, it revealed that cell concentration became higher, and residue content of the mixed substances was lower in a week. We can draw a conclusion that the mixture can promote PSB reproduction and protein content.
引文
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