代谢工程方法改造谷氨酸棒状杆菌生产乙偶姻  

Metabolic Engineering of Corynebacterium Glutamicum for Acetoin Production

作　　者：马红武[1,2] 刘会娟[1] 朱年青[1] 陈涛[1] 

机构地区：[1]天津大学化工学院,天津300072 [2]中科院天津工业生物技术研究所,天津300308

出　　处：《天津大学学报（自然科学与工程技术版）》2014年第11期967-972,共6页Journal of Tianjin University：Science and Technology

基　　金：国家重点基础研究发展计划(973计划)资助项目(2011CBA00804;2012CB725203);国家自然科学基金资助项目(21176182)

摘　　要：应用代谢工程方法对Corynebacterium glutamicum ATCC 13032生物合成乙偶姻进行了研究.在C.glutamicum ATCC 13032中导入了Bacillus subtilis 168的乙偶姻合成途径的相关基因als SD操纵子,工程菌株的乙偶姻产量为2.14,g/L.为了增加合成乙偶姻的直接前体物丙酮酸的供给,进一步敲除了丙酮酸脱氢酶复合体E1亚基的编码基因(ace E)和乳酸脱氢酶基因(ldh A),工程菌株的乙偶姻产量提高到5.09,g/L.最后,敲除了工程菌株的2,3-丁二醇脱氢酶基因(but A)以阻断副产物2,3-丁二醇的合成,在优化的溶氧条件下,菌株CGL3在基本培养基中乙偶姻产量提高到8.33,g/L,达到理论得率的51.5%.实验结果表明经过代谢工程改造的C.glutamicum ATCC 13032具有良好的乙偶姻合成能力和应用潜力.The acetoin was synthesized using metabolic engineering method from Corynebacterium glutamicumATCC 13032. The alsSD operon derived from Bacillus subtilis 168 was expressed in C. glutamicum ATCC 13032 toyield acetoin of 2.14,g/L in the engineering strain. Additional inactivation of aceE gene encoding the E1p enzyme ofthe pyruvate dehydrogenase complex and ldhA gene encoding NAD＋-dependent L-lactate dehydrogenase increased thesupply of acetoin precursor. The acetoin production increased to 5.09,g/L with the newly constructed strain. Subsequently,formation of 2,3-butanediol was inhibited by deleting butA gene encoding for 2,3-butanediol dehydrogenase.Total yield of acetoin reached 8.33,g/L（51.1% of theoretical yield）under the optimum fermentation conditionin the strain CGL3. The experimental results show that the strain of C. glutamicum ATCC 13032,engineered metabolicallycan provide a potential application for acetoin synthesis.

关 键 词：代谢工程 乙偶姻 谷氨酸棒状杆菌 乙酰乳酸合成酶 乙酰乳酸脱羧酶 

分 类 号：Q815[生物学—生物工程]