关键词: 谷氨酸棒状杆菌, 葡萄糖, 基因敲除

Abstract: Corynebacterium glutamicum is an important production strains, which is used to produce glutamic acid, lysine and glutamine in areas of flavor enhancer and animal feed additives. Corynebacterium glutamicum, whose metabolism is related to productivity and costs. Therefore, to study the regulation of glucose transport systems and molecular level has an importantvalue. In this study, we use reverse metabolic engineering strategies as a starting strain of wild type C. glutamicum ATCC 13032 by homologous recombination without resistance marker, knocked encoding glucose transporter gene critical systems pts ptsG, ptsH-ptsI and glucose transport system critical transporter gene abc, abc2 and iolt1, got five successive knockout mutants corresponding gene. Experimental results show that: compared to the wild-type, the growth rate and Glucose consumption of CGΔptsG was 50% and 77.7%; the growth rate and Glucose consumption of CGΔptsH-ptsI was 39.5% and 66.8%; the growth rate and Glucose consumption of CGΔabc was 36% and 60%; the growth rate and Glucose consumption of CGΔabc2 was 26.2% and 30.1%; the growth rate and Glucose consumption of CGΔiolt1 was 0, the glucose metabolism of Corynebacterium glutamicum is blocked. It turned out that the glucose transporter function was controlled by ptsG、ptsH-ptsI、abc、abc2 and iolt1 genes encoding transporter proteins. This research not only laid the theoretical foundation for molecular level study of glucose transport and metabolism, but also provides a basic genetic resources for the common fermentation of Corynebacterium glutamicum and other glucose utilization microorganisms.

Key words: Corynebacterium glutamicum, glucose, gene knockout