NADH-driven poly-3-hydroxybutyrate accumulation in Escherichia coli

NADH-driven poly-3-hydroxybutyrate accumulation in Escherichia coli: Data from enzymatic assays and oxygen-limited continuous cultures

Olavarria, Karel (TU Delft BT/Industriele Microbiologie)
Quakkelaar, Caspar (Student TU Delft)
van Renselaar, Joachim (Student TU Delft)
Langerak, D. (TU Delft OLD BT/Cell Systems Engineering)
van Loosdrecht, Mark C.M. (TU Delft BT/Environmental Biotechnology)
Wahl, S.A. (TU Delft BT/Industriele Microbiologie)

2020

Biosynthesis of poly-3-hydroxybutyrate (PHB) as a fermentation product enables the coupling of growth and product generation. Moreover, the reduction of oxygen supply should reduce operative cost and increase product yield. Generation of PHB as a fermentation product depends on the in vivo activity of an NADH-preferring acetoacetyl-CoA reductase. Proof of this concept requires (i) quantification of the cofactor preference, in physiologically relevant conditions, of a putative NADH-preferring acetoacetyl-CoA reductase and (ii) verification of PHB accumulation using an NADH-preferring acetoacetyl-CoA reductase in a species naturally incapable of doing so, for example, Escherichia coli. This dataset contains kinetic data obtained by spectrophotometry and data from a continuous culture of an engineered E. coli strain accumulating PHB under oxygen-limiting conditions. In this dataset it is possible to find (1) enzyme stability assays; (2) initial rates and progress curves from reactions catalyzed by two acetoacetyl-CoA reductases; (3) estimations of the relative use of NADH and NADPH by two acetoacetyl-CoA reductases; (4) estimations of the flux capacity of the reaction catalyzed by an acetoacetyl-CoA reductase; (5) biomass composition of an engineered E. coli strain transformed with a plasmid; (6) calculation of reconciled specific rates of this engineered strain growing on sucrose as the sole carbon source under oxygen limitation and (7) metabolic fluxes distributions during the continuous growth of this engineered strain. Because a relatively small number of acetoacetyl-CoA reductases have been kinetically characterized, data and scripts here provided could be useful for further kinetic characterizations. Moreover, the procedure described to estimate biomass composition could be interesting to estimate plasmid and protein burden in other strains. Application of data reconciliation to fermentations should help to obtain specific rates consistent with the principle of mass and electron conservation. All the required data and scripts to perform these analyses are deposited in a Mendeley Data repository. This article was co-submitted with the manuscript entitled “An NADH preferring acetoacetyl-CoA reductase is engaged in poly-3-hydroxybutyrate accumulation in Escherichiasia. coli”.

Acetoacetyl-CoA reductase
Candidatus Accumulibacter phosphatis
Cofactor preference
Continuous culture
Data reconciliation
Kinetic characterization
Metabolic fluxes
Polyhydroxybutyrate

http://resolver.tudelft.nl/uuid:a6fd076d-e012-4bef-b862-d64657b3ba57

https://doi.org/10.1016/j.dib.2020.106588

2352-3409

Data in Brief, 33

Institutional Repository

journal article

© 2020 Karel Olavarria, Caspar Quakkelaar, Joachim van Renselaar, D. Langerak, Mark C.M. van Loosdrecht, S.A. Wahl