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San, Ka-Yiu Bennett, George N. Gonzalez, Ramon Jan, Joanna 2012-09-06T04:18:07Z 2012-09-06T04:18:10Z 2012-09-06T04:18:07Z 2012-09-06T04:18:10Z 2012-05 2012-09-05 May 2012 2012-09-06T04:18:10Z Jan, Joanna. "Metabolic Engineering and Transhydrogenase Effects on NADPH Availability in Escherichia coli." (2012) Master’s Thesis, Rice University. <a href="https://hdl.handle.net/1911/64664">https://hdl.handle.net/1911/64664</a>. https://hdl.handle.net/1911/64664 123456789/ETD-2012-05-127 The ultimate goal in the field of metabolic engineering is improving cellular processes in a rational manner using engineering design principles and molecular biology techniques. The syntheses of several industrially useful compounds are cofactor-dependent. The reducing equivalent NADPH is required in several enzymatic reactions leading up to the synthesis of high-value compounds like polymers, chiral alcohols, and antibiotics. However, it’s a highly costly compound with limited intracellular availability. This study focuses on the genetic manipulation of a whole-cell system using the two transhydrogenase isoforms pntAB and udhA. Two model systems are used: 1) the production of (S)-2-chloropropionate and 2) the production of poly(3-hydroxybutyrate). Results suggest that the presence of udhA increases product yield and NADPH availability while the presence of pntAB has the opposite effect. A maximum product yield of 1.4 mole-product/mole-glucose was achieved aerobically in a pntAB-deletion strain with udhA overexpression, a 150% improvement over the wild-type control strain. application/pdf eng Copyright is held by the author, unless otherwise indicated. Permission to reuse, publish, or reproduce the work beyond the bounds of fair use or other exemptions to copyright law must be obtained from the copyright holder. Metabolic engineering Escherichia coli Nadph Cofactor Transhydrogenase Metabolic Engineering and Transhydrogenase Effects on NADPH Availability in Escherichia coli Thesis Text Bioengineering Engineering Rice University Masters Master of Science open.access