2017 ; Vol.27-10: 1867~1876
|Author||Xue-Mei Jin, Yong-Keun Chang, Jae Hag Lee, Soon-Kwang Hong|
|Place of duty||Department of Biological Science and Bioinformatics, Myongji University, Yongin 17058, Republic of Korea|
|Title||Effects of Increased NADPH Concentration by Metabolic Engineering of the Pentose Phosphate Pathway on Antibiotic Production and Sporulation in Streptomyces lividans TK24|
J. Microbiol. Biotechnol.2017 ;
|Abstract||Most of the biosynthetic pathways for secondary metabolites are influenced by carbon
metabolism and supply of cytosolic NADPH. We engineered carbon distribution to the
pentose phosphate pathway (PPP) and redesigned the host to produce high levels of NADPH
and primary intermediates from the PPP. The main enzymes producing NADPH in the PPP,
glucose 6-phosphate dehydrogenase (encoded by zwf1 and zwf2) and 6-phosphogluconate
dehydrogenase (encoded by zwf3), were overexpressed with opc encoding a positive allosteric
effector essential for Zwf activity in various combinations in Streptomyces lividans TK24. Most
S. lividans transformants showed better cell growth and higher concentration of cytosolic
NADPH than those of the control, and S. lividans TK24/pWHM3-Z23O2 containing
zwf2+zwf3+opc2 showed the highest NADPH concentration but poor sporulation in R2YE
medium. S. lividans TK24/pWHM3-Z23O2 in minimal medium showed the maximum growth
(6.2 mg/ml) at day 4. Thereafter, a gradual decrease of biomass and a sharp increase of
cytosolic NADPH and sedoheptulose 7-phosphate between days 2 and 4 and between days 1
and 3, respectively, were observed. Moreover, S. lividans TK24/pWHM3-Z23O2 produced 0.9
times less actinorhodin but 1.8 times more undecylprodigiosin than the control. These results
suggested that the increased NADPH concentration and various intermediates from the PPP
specifically triggered undecylprodigiosin biosynthesis that required many precursors and
NADPH-dependent reduction reaction. This study is the first report on bespoke metabolic
engineering of PPP routes especially suitable for producing secondary metabolites that need
diverse primary precursors and NADPH, which is useful information for metabolic
engineering in Streptomyces.|
|Key_word||Glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, actinorhodin, undecylprodigiosin, Streptomyces lividans, pentose phosphate pathway|
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