Improvement of NADPH bioavailability in Escherichia coli through the use of phosphofructokinase deficient strains |
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Authors: | Yipeng Wang Ka-Yiu San George N. Bennett |
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Affiliation: | 1. Department of Biochemistry and Cell Biology, MS-140, Rice University, 6100 Main Street, Houston, TX, 77005-1892, USA 2. Department of Bioengineering, Rice University, Houston, TX, 77005, USA 3. Department of Chemical and Biomolecular Engineering, Rice University, Houston, TX, 77251, USA
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Abstract: | NADPH-dependent reactions play important roles in production of industrially valuable compounds. In this study, we used phosphofructokinase (PFK)-deficient strains to direct fructose-6-phosphate to be oxidized through the pentose phosphate pathway (PPP) to increase NADPH generation. pfkA or pfkB single deletion and double-deletion strains were tested for their ability to produce lycopene. Since lycopene biosynthesis requires many NADPH, levels of lycopene were compared in a set of isogenic strains, with the pfkA single deletion strain showing the highest lycopene yield. Using another NADPH-requiring process, a one-step reduction reaction of 2-chloroacrylate to 2-chloropropionic acid by 2-haloacrylate reductase, the pfkA pfkB double-deletion strain showed the highest yield of 2-chloropropionic acid product. The combined effect of glucose-6-phosphate dehydrogenase overexpression or lactate dehydrogenase deletion with PFK deficiency on NADPH bioavailability was also studied. The results indicated that the flux distribution of fructose-6-phosphate between glycolysis and the pentose phosphate pathway determines the amount of NAPDH available for reductive biosynthesis. |
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