探索一种新型生物发酵途径生产乙醇  

A Novel Method of Ethanol Fermentation from Biomass Resources

作　　者：黄立[1] 黄潇[1] 董元楷[1] 邢芸[1] 葛驰宇[1] 刘景晶[1] 曹荣月[1] 

机构地区：[1]中国药科大学生命科学与技术学院天然药物活性物质与功能国家重点实验室,江苏南京210009

出　　处：《药物生物技术》2012年第3期213-217,共5页Pharmaceutical Biotechnology

基　　金：国家自然科学基金(30872393;811729731);先声药业创新基金(CX10B-004XS)

摘　　要：在质粒pUC19中插入菊欧氏杆菌pehX启动子,并与全合成的运动发酵单胞菌的丙酮酸脱羧酶基因(pdc),乙醇脱氢酶基因(adhB)串联,构建pUC19-pdc-adhB乙醇发酵重组克隆,转化大肠杆菌DH5α,经气相色谱检测,在大肠杆菌中乙醇的表达量达到0.85%。菊欧氏杆菌富含多种纤维素酶,与一些仅可利用基本糖类作为碳源的大肠杆菌等相比,其可以利用纤维素作为碳源。本实验设计将pUC19-pdc-adhB乙醇发酵重组克隆转化至菊欧氏杆菌,尝试用氯化钙、电击等转化方法,但均没有获得转化子。提示需要寻找新的转化方法或者将pdc和adhB基因整合到菊欧氏杆菌的基因组中以使其能产生乙醇。The clone of ethanol fermentation was constructed by pUC19 which carried pehX promoter from Envinia chrysanthemi and codon-replaced pyruvate decarboxylase gene（pdc） and alcohol dehydrogenase gene（adhB） from Zymomonas mobilis. After the new constructed clone was transformed into E. coli DH5α, the ethanol content of the E. coli DH5c~ broth reaches 0.85% through gas chromatography detection. The clone was transformed into Erwinia chrysanthemi which is a cellulase-rich kind of plant pathogens which can easily use cellulose as a carbon source like tree bark or duckweed while most kinds of other bacteria only use some normal carbon sources like glucose or saccharose. However, a variety of methods and conditions including the measure of CaC12 and electro- poration were tested to transform the clone into Erwinia chrysanthemi. They ultimately got limited transformants. The results suggested that new methods need to be established to transform the clone into Erwinia chrysanthemi or directly import the pdc and adhB into the genome of Erwinia chrysanthemi to construct a new breed of bacteria which can produce ethanol for commercial practice.

关 键 词：菊欧氏杆菌 丙酮酸脱羧酶基因(pdc) 乙醇脱氢酶基因(adhB) 气相色谱 转化 

分 类 号：Q78[生物学—分子生物学]