高产生物丁新菌株的筛选、鉴定及发酵研究.docVIP

高产生物丁醇新菌株的筛选、鉴定及发酵研究 裴建新1左文朴1庞浩1黄志1，黎贞崇1黄日波1，2 （1广西科学院 国家非粮生物质能源工程技术研究中心广西 南宁 .广西大学微生物及植物遗传工程教育部重点实验室，广西 南宁 530003） 摘要：为了获得高效生产丁醇的菌株，通过玉米培养基富集丁醇耐受筛选从自然环境（牛粪沼气池土壤）60份样品中分离获得的新菌株，菌株命名为gxzp-13-2。经浓度葡萄糖发酵验表明5%葡萄糖发酵丁醇产量较高，最高可达10.05 g/L，总溶剂达到13.95 g/L，丁醇比约72.1%，转化率为3.8 %。对此菌株进行了分子鉴定，经16S rDNA基因序列同源性分析gxzp-13-2与拜氏梭菌（Clostridium beijerinckii），gxzp-13-2在厌氧固体培养基上生长，菌落显紫色。该菌株为下一步的菌种基因改造工作提供了研究基础，也为丁醇发酵提供宝贵的菌种资源。关键词：拜氏梭菌生物丁醇鉴定发酵Screening and Identification of a Novel Strains for Producing butanol PEI Jian-xin1，ZUO Wen-pu1，PANG Hao1，ZHANG Sui-sheng1，HUANG Zhi-min1， LI Zhen-chong1，HUANG Ri-bo1, 2 （1. National Engineering Research Center for Non-food Biorefinery，Guangxi Academy of Science， Nanning 530007，China；2. Key Laboratory of Microbial and Plant Genetic Engineering of Ministry of Education，Guangxi University，Nanning 530003， China） Abstract: Butanol has great potential to be renewable sources of energy. In order to obtain efficient strains of producing butanol, one purple bacterial colony was screened by enriching on corn medium and selecting tolerant of butanol from 60 samples，which were got from cow dung, methanetank and soil. The new strain could effectively produce butanol，which was named as gxzp-13-2. The results of fermentation by different concentration of glucose showed: the yield of butanol fermented by 5% of glucose were the highest, reached 10.05g/L, the total solvent, rate of butanol and conversion of which were 13.95 g/L, 72.1 % and 31.8 % respectively. It had the highest homology with Clostridium beijerinckii through 16S rDNA sequence analysis，and their similarity reached 94%. When gxzp-13-2 grew on anaerobic medium，it appeared purple. This new strain could not only provide basis for alternating genes of strains for further study, but also provide valuable stains resource for fermentation butanol. Key words: Clostridium beijerinckii；biobutanol；identification；fermentation 0 引言 由于丁醇的能量密度高，亲水性低，蒸汽压低，辛烷值高等特点，被认为是最有发展潜力的新型液体燃料之一[1]。传统的丁醇生产方法主要是以石油为原料经过羰基合成或醇醛缩合而成