产氢菌的筛选及培养优化的研究

【作者】 赵小尊；

【导师】 范代娣；

【作者基本信息】 西北大学 ， 化学工程， 2009， 硕士

【摘要】 在21世纪,清洁的氢能是最有发展前景的替代能源之一。当前,氢气的开发和利用已成为科学研究的重点课题之一。生物制氢技术因其具有清洁、节能和不消耗矿石资源等突出优点而备受人们关注。本论文定位于产氢菌的筛选及培养优化的研究。实验挑选了5种来源不同的污泥样品,对其进行厌氧梯度驯化,分析比较了不同样品产氢能力的差别。其中来源于深层河底的污泥样品产氢能力达1.5mol H₂/（mol glucose）。同时从上述产氢能力高的污泥样品中筛选出了5株产氢能力较好的菌种,以CH1的产氢能力最高。对该菌株进行了形态观察和一些基本的生理生化试验,初步确定菌株的特性为:革兰氏染色阴性,无荚膜、芽孢,穿刺培养后,菌落长如根状,判断该菌运动性较强,估计有鞭毛。葡萄糖发酵试验为双阳性（产气并产酸）;甲基红试验为阳性;V.P.试验阳性;吲哚试验阴性;产H₂S;具有还原硝酸盐的能力。又通过扫描电镜观察,该菌为杆状,菌体大小约1.5～2.5μm*0.5μm。并进一步通过16S rDNA序列分析。初步鉴定该菌为产气肠杆菌。对该菌进行生长特性研究表明:CH1的产氢过程是和其生长过程密切相关,氢气是其生长伴随型代谢产物。该菌种的最佳培养温度是35℃,最适的pH值为6。最适的碳源是葡萄糖,最佳的氮源是蛋白胨、酵母粉和牛肉膏。同时CH1发酵周期很短,只有12h。更多还原

【Abstract】 In the 21 century,clean hydrogen energy is one of the most perspective replaceable energy resources.The development and utilization of hydrogen is becoming one of the most important issues in scientific researches.Biological hydrogen,one of hydrogen productions,has attracted widespread attentions for its clean,economic and not consuming any fossil fuels.In this paper,five different sledges are chosen for anaerobic gradient acclimatization.The sample（3） with the highest biohydrogen production capacity and the flocculation ratio,which is 1.5mol H₂/（mol glucose）;at the same time,five strains with higher efficiency are obtained among those sledges,especially CH1, which is the best of all.It was initiative identification by morphological and biochemical analysis:Bacteria Gram stain-negative,non-Clostridium perfringens, Bacillus,puncture cultured long as the root-like colony,judge the movement of more bacteria,it is estimated that flagellin.Glucose fermentation test for the double-positive（and acid gas）;methyl red test was positive;VP test positive;Indole test negative;producing H₂S.It also has the ability to restore nitrate.By scanning electron microscopy,the strain was the rod-shaped bacteria with the size of about 1.5～2.5μm ^*0.5μm.And the strain was determined as Enterobacter aerogenes through the sequence analysis 16sRNA Preliminary identification.From the research of the bacterium,we can know that:the hydrogen production was associated with the growth of the bacterium,which was a accompany production. The suitable condition for the bacterium was that:35℃,pH 6.the best carbon source and nitrogen source for the bacteria were glucose and mixture organic nitrogen which were the mixture of Yeast,peptone and beef extract,at the same time,CH1 has a short cycle which only last for 12 hours.更多还原