RT journal article
T1 Metabolomics of Escherichia coli for Disclosing Novel Metabolic Engineering Strategies for Enhancing Hydrogen and Ethanol Production
A1 Valle Gallardo, Antonio
A1 Calle Sierra, María Elena de la
A1 Muhamadali, Howbeer
A1 Hollywood, Katherine A.
A1 Xu, Yun
A1 Lloyd, Jonathan R.
A1 Goodacre, Royston
A1 Cantero Moreno, Domingo
A1 Bolívar Pérez, Jorge
A2 BiomedicinaBiotecnología y Salud Pública
A2 Ingeniería Química y Tecnología de Alimentos
K1 Escherichia coli
K1 ethanol
K1 FT-IR
K1 GC-MS
K1 hydrogen
K1 metabolomics
AB The biological production of hydrogen is an appealing approach to mitigating the environmental problems caused by the diminishing supply of fossil fuels and the need for greener energy. Escherichia coli is one of the best-characterized microorganisms capable of consuming glycerol—a waste product of the biodiesel industry—and producing H2 and ethanol. However, the natural capacity of E. coli to generate these compounds is insufficient for commercial or industrial purposes. Metabolic engineering allows for the rewiring of the carbon source towards H2 production, although the strategies for achieving this aim are difficult to foresee. In this work, we use metabolomics platforms through GC-MS and FT-IR techniques to detect metabolic bottlenecks in the engineered ΔldhΔgndΔfrdBC::kan (M4) and ΔldhΔgndΔfrdBCΔtdcE::kan (M5) E. coli strains, previously reported as improved H2 and ethanol producers. In the M5 strain, increased intracellular citrate and malate were detected by GC-MS. These metabolites can be redirected towards acetyl-CoA and formate by the overexpression of the citrate lyase (CIT) enzyme and by co-overexpressing the anaplerotic human phosphoenol pyruvate carboxykinase (hPEPCK) or malic (MaeA) enzymes using inducible promoter vectors. These strategies enhanced specific H2 production by up to 1.25- and 1.49-fold, respectively, compared to the reference strains. Other parameters, such as ethanol and H2 yields, were also enhanced. However, these vectors may provoke metabolic burden in anaerobic conditions. Therefore, alternative strategies for a tighter control of protein expression should be addressed in order to avoid undesirable effects in the metabolic network.
PB MDPI
SN 1422-0067
YR 2023
FD 2023
LK http://hdl.handle.net/10498/32494
UL http://hdl.handle.net/10498/32494
LA eng
DS Repositorio Institucional de la Universidad de Cádiz
RD 07-nov-2026