发酵法产生物丁醇的研究

【作者】 于鑫；

【导师】 姚淑敏； 华栋梁；

【作者基本信息】 曲阜师范大学 ， 生物化学与分子生物学， 2014， 硕士

【摘要】 众所周知，能源一直是一个国家的经济和社会发展和进步所必需的一个重要物质基础。虽然传统的化石燃料是现代工业的命脉，但是由于石油等化石资源的日益枯竭，价格不断地攀升，以及其所引起的严重的环境污染，使得人们必须转向寻找新的可再生环保能源生物能源。生物能源是一种可再生的绿色能源，即指使用一些可再生和环境友好的生物质，如玉米、薯类、农作物秸秆等为原料生产的乙醇、丁醇和柴油等石油替代燃料。开发和发展生物能源，既能有效控制环境污染，减轻对石油资源的依赖，同时又能促进农业产业链的发展，被认为是解决世界能源危机的最理想途径之一。燃料乙醇生产技术已经很成熟，并已经进行工业化生产。但是乙醇具有腐蚀运输管道等缺点。而生物丁醇作为能够合成多种有机物的重要平台化合物，和能与汽油以任意比例混溶等较其他能源更具优势的液体生物燃料，已经逐渐被各国重视和并得到广泛的研究。但是在生物丁醇的发展过程中，仍然存在着一些问题没有得到很好的解决。比如，丁醇对细胞的毒性，导致产量达不到很高的值。本文对丙酮丁醇梭菌（Clostridium acetobutylicum）发酵产生物丁醇的过程一些环节作了研究。重在研究树脂吸附的新型原位发酵技术和深入研究原位萃取技术，以解决丁醇毒性和提高产量。首先对丁醇丙酮梭菌培养条件进行了简单的优化和研究；第二利用丁醇水溶液对树脂和萃取剂做了筛选和特性研究；第三将筛选的树脂和萃取剂进行原位发酵研究。通过实验，我们确定了最佳的发酵葡萄糖浓度是60g/l。另外，酵母粉添加对发酵产量影响不大，对种子液热激可提高发酵产量。利用丁醇水溶液，筛选到对丁醇吸附率最高的大孔吸附树脂DM11，初步验证了树脂原位吸附发酵的可行性。对其添加比例等进行研究之后，进行原位发酵实验。实验表明：树脂的添加对提高丁醇产量有一定的效果。对各种萃取剂研究后发现，正庚醇分配率最大。其原位发酵实验表明：添加正庚醇后，丁醇浓度增加了14%。更多还原

【Abstract】 As we all know, energy is a significant material foundation to the development of theeconomy and society for a country. The traditional fossil fuel is the lifeblood of modern industry.However, people must turn to find new renewable and environmental energy—biofuel, as Oiland other fossil resources are exhausting and the price is rising day by day. Biofuel, a kind ofgreen energy, is an alternative fuel of petroleum using some renewable and eco-friendly biomasssuch as corn, as materials to produce bioethanol, biodiesel, etc. Developing biofuel can controlenvironment pollution efficiently, as well as reduce the dependence on oil resources and drivethe development of agricultural industry chain at the same time. So it is considered to be one ofthe most ideal ways to solve the global energy crisis.The production technology of bioethanol has been very mature. It has realized theindustrialized production. But it has some disadvantages such as pipeline corrosion. While as animportant platform chemical compound that can synthesize various organics and blending withgasoline in any proportion, biobutanol is a kind of liquid fuel that has more advantages thansome other energy. So many countries have paid attention to and extensively researched it.During the development process of biobutanol, however, there are still some problems thathaven’t been solved. For example, the toxicity of butanol to cell leads to low yield.This paper studied some steps during the fermentation process of Clostridiumacetobutylicum producing biobutanol. The research focused on the new in-situ fermentationtechnology of resin adsorption and lucubrating the technology of in-situ extraction, so as to solvethe toxicity of butanol and improve its yield. First, the culture conditions of Clostridiumacetobutylicum were optimized and researched simply. Second, resins and extraction agent werescreened and researched using butanol aqueous solution. Third, the resin and extractationscreened were used for the in-situ fermentation. Finally, the fermentation process andcharacteristics were studied further through the fermentation in small fermentation tank.By the experiments, the optimal fermentation concentration of glucose was determined to be60g/l. The addition of yeast powder in medium had little influence on fermentation yield, whileheat shock treatment for seed solution could improve the yield. The adsorption rate ofmacroreticular absorbent DM11that screened by butanol aqueous solution was highest. Itverified the feasibility of resin in-situ adsorption fermentation primarily. After researching theaddition rate and so on, the in-situ fermentation using resin DM11was carried out. Theexperiment indicated that the addition of resin had certain effect on the production of butanol.After the research on various extraction agent, the distribution rate of n-heptanol was found to bethe maximum. The in-situ fermentation experiment using n-heptanol indicated that theconcentration of butanol had been increased by fourteen percent.更多还原