Metabolic flux control at the pyruvate node in an anaerobic Escherichia coli strain with an active pyruvate dehydrogenase.

Applied and Environmental Microbiology
Qingzhao WangK T Shanmugam

Abstract

During anaerobic growth of Escherichia coli, pyruvate formate-lyase (PFL) and lactate dehydrogenase (LDH) channel pyruvate toward a mixture of fermentation products. We have introduced a third branch at the pyruvate node in a mutant of E. coli with a mutation in pyruvate dehydrogenase (PDH*) that renders the enzyme less sensitive to inhibition by NADH. The key starting enzymes of the three branches at the pyruvate node in such a mutant, PDH*, PFL, and LDH, have different metabolic potentials and kinetic properties. In such a mutant (strain QZ2), pyruvate flux through LDH was about 30%, with the remainder of the flux occurring through PFL, indicating that LDH is a preferred route of pyruvate conversion over PDH*. In a pfl mutant (strain YK167) with both PDH* and LDH activities, flux through PDH* was about 33% of the total, confirming the ability of LDH to outcompete the PDH pathway for pyruvate in vivo. Only in the absence of LDH (strain QZ3) was pyruvate carbon equally distributed between the PDH* and PFL pathways. A pfl mutant with LDH and PDH* activities, as well as a pfl ldh double mutant with PDH* activity, had a surprisingly low cell yield per mole of ATP (Y(ATP)) (about 7.0 g of cells per mol of ATP) compared to 10.9 g of...Continue Reading

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Citations

Mar 1, 2012·Journal of Industrial Microbiology & Biotechnology·Ryan ManowShengde Zhou
Aug 15, 2013·Journal of Biotechnology·Amanda HildebrandZhiliang Fan
Jan 11, 2012·Biotechnology and Bioengineering·Qingzhao WangJauhleene Chamu
Apr 19, 2015·Microbial Cell Factories·Stephen G WuYinjie J Tang
Aug 25, 2016·Journal of the American Chemical Society·Piotr DzienLucio Frydman
Nov 2, 2016·Journal of Industrial Microbiology & Biotechnology·George N Bennett, Ka-Yiu San
May 31, 2019·Applied Microbiology and Biotechnology·Karel OlavarriaSebastian Aljoscha Wahl
Apr 29, 2020·Metabolites·Jason T KuEthan I Lan
Sep 29, 2011·Prikladnaia biokhimiia i mikrobiologiia·A Iu SkorokhodovaV G Debabov
May 5, 2017·Scientific Reports·Stefan BehrKirsten Jung
Feb 9, 2021·Metabolic Engineering Communications·David C GarciaMitchel J Doktycz
Jul 22, 2021·The ISME Journal·Angeliki MarietouBo Barker Jørgensen

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