代谢工程改造谷氨酸棒杆菌生成丙酮酸.pdf

生物技术通报 ·研究报告· BIOTECHNOLOGY BULLETIN ( ) 2016, 32 8 :226-232 代谢工程改造谷氨酸棒杆菌生成丙酮酸 于莹  许湄雪  刘金雷  范荣  冯惠勇  李天明  （河北科技大学，石家庄 050018） 摘 要 ： 旨在增加谷氨酸棒杆菌代谢过程中丙酮酸的积累、减少副产物的生成，利用同源重组的方法，敲除了丙酮酸代谢 过程中支流代谢途径的关键基因 ：丙酮酸醌氧化还原酶 基因、丙酮酸脱氢酶 基因和乳酸脱氢酶 基因，获得了基因缺 pqo pdh lldh 失工程菌株。利用4.5% 的葡萄糖复合培养基，工程菌经摇瓶发酵48 h， 丙酮酸的浓度达到14.6 g/L，而野生菌株仅为0.45 g/L ；工 程菌的糖酸转化率为33.18%， 比野生菌提高了32.5 倍。 关键词 ： 丙酮酸；谷氨酸棒状杆菌 ；丙酮酸醌氧化还原酶；丙酮酸脱氢酶；乳酸脱氢酶 DOI ：10.13560/ki.biotech.bull.1985.2016.08.033 Metabolic Engineering for Modifying Corynebacterium glutamicum to Produce More Pyruvate YU Ying XU Mei-xue LIU Jin-lei FAN Rong FENG Hui-yong LI Tian-ming （Hebei University of Science and Technology ，Shijiazhuang 050018 ） Abstract: In order to increase the accumulation of pyruvate while decrease the production of by-products during the metabolic process of Corynebacterium glutamicum ，the homologous recombination was employed to knock out the key genes of pyruvate ：quinone oxidoreductase （ ），pyruvate dehydrogenase（ ）and L-lactate dehydrogenase（ ）in the tributary metabolic pathways，thus the gene deletion pqo pdh lldh mutation strains were constructed. Using compound medium with 4.5% glucose as carbon sources，the concentration of pyruvate with mutation strain at shake flask fermentation for 48 hours reached 14.6 g/L，while that with the wild type only was 0.45 g/L. The conversion rate of sugar to acid with the mutation strain was 33.18% and 32.5 times higher than the wild type.