The pentose phosphate pathway (PPP) and glycolysis comprise the most central metabolic pathways in the primary metabolism of Streptomyces. Most of the antibiotic biosynthetic pathways include reductive steps where NADPH is required as a reducing factor. NADPH is produced in PPP, by glucose-6-phosphate dehydrogenase (Zwf1 and Zwf2) and 6- phosphogluconate dehydrogenase (Zwf3). According to the report, the two genes (zwf1 and zwf2) were translationally coupled with the downstream opc1 and opc2 genes, respectively. The opc1 and opc2 genes were reported to be essential for Zwf activity in Cyanobacteria and corynebacterium. On the basis of the rationale, we postulated that the increase of NADPH concentration in the microbial cells would result in overproduction of secondary metabolites. Based on the genomic sequence of S. coelicolor, firstly, four different combinations of zwf and opc genes, were introduced into the pUWL201PW vector, and the resulting vectors were transformed into Streptomyces lividans TK24. Secondly, the carbon metabolism of S. lividans transformanrs was investigated. Especially, biomass, antibiotics, NADPH, and sedoheptulose- 7-phosphate production in the engineered cells were nalyzed. Taken as a whole, our results indicate engineering of PPP by the glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase genes is a useful way to elevate intracellular concentration of NADPH and sedoheptulose-7-phosphate as well as biomass, which may be applied to improve productivity of secondary metabolites biosynthesized from the intermediates of PPP. [supported by grant no. 2009-0073015 from the Basic Research Program of the National Research Foundation]
한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
설립연도
1984
분야
공학>생물공학
소개
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