ArticleApplied and environmental microbiology2008
NADH availability limits asymmetric biocatalytic epoxidation in a growing recombinant Escherichia coli strain.
Article in Applied and environmental microbiology, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed, 76 citations in OpenAlex.
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- Exogenous NADH enhances polycyclic aromatic hydrocarbon degradation inApplied and environmental microbiology · 2026Article
- Strategies found not to be suitable for stabilizing high steroid hydroxylation activities of CYP450 BM3-based whole-cell biocatalysts.PloS one · 2024Article
- Exploitation of Hetero- and Phototrophic Metabolic Modules for Redox-Intensive Whole-Cell Biocatalysis.Frontiers in bioengineering and biotechnology · 2022Review
- Cytochrome P450-mediated N-demethylation of noscapine by whole-cell biotransformation: process limitations and strategies for optimisation.Journal of industrial microbiology & biotechnology · 2020Article
- Metabolic response ofEngineering in life sciences · 2017Article
- The CreC Regulator of Escherichia coli, a New Target for Metabolic Manipulations.Applied and environmental microbiology · 2016Article
- The dynamic influence of cells on the formation of stable emulsions in organic-aqueous biotransformations.Journal of industrial microbiology & biotechnology · 2015Article
- Variability in subpopulation formation propagates into biocatalytic variability of engineered Pseudomonas putida strains.Frontiers in microbiology · 2015Article
- Flavoprotein monooxygenases for oxidative biocatalysis: recombinant expression in microbial hosts and applications.Frontiers in microbiology · 2014Review
- Engineering NAD+ availability for Escherichia coli whole-cell biocatalysis: a case study for dihydroxyacetone production.Microbial cell factories · 2013Article
- Proline availability regulates proline-4-hydroxylase synthesis and substrate uptake in proline-hydroxylating recombinant Escherichia coli.Applied and environmental microbiology · 2013Article
- A stepwise approach for the reproducible optimization of PAMO expression in Escherichia coli for whole-cell biocatalysis.BMC biotechnology · 2012Article
- Genome-scale modeling using flux ratio constraints to enable metabolic engineering of clostridial metabolism in silico.BMC systems biology · 2012Article
- Determining the extremes of the cellular NAD(H) level by using an Escherichia coli NAD(+)-auxotrophic mutant.Applied and environmental microbiology · 2011Article
- Regioselective aromatic hydroxylation of quinaldine by water using quinaldine 4-oxidase in recombinant Pseudomonas putida.Journal of industrial microbiology & biotechnology · 2011Article
- Anaerobic obligatory xylitol production in Escherichia coli strains devoid of native fermentation pathways.Applied and environmental microbiology · 2011Article
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Authors and funding
4 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Styrene can efficiently be oxidized to (S)-styrene oxide by recombinant Escherichia coli expressing the styrene monooxygenase genes styAB from Pseudomonas sp. strain VLB120. Targeting microbial physiology during whole-cell redox biocatalysis, we investigated the interdependency of styrene epoxidation, growth, and carbon metabolism on the basis of mass balances obtained from continuous two-liquid-phase cultures. Full induction of styAB expression led to growth inhibition, which could be attenuated by reducing expression levels. Operation at subtoxic substrate and product concentrations and variation of the epoxidation rate via the styrene feed concentration allowed a detailed analysis of carbon metabolism and bioconversion kinetics. Fine-tuned styAB expression and increasing specific epoxidation rates resulted in decreasing biomass yields, increasing specific rates for glucose uptake and the tricarboxylic acid (TCA) cycle, and finally saturation of the TCA cycle and acetate formation. Interestingly, the biocatalysis-related NAD(P)H consumption was 3.2 to 3.7 times higher than expected from the epoxidation stoichiometry. Possible reasons include uncoupling of styrene epoxidation and NADH oxidation and increased maintenance requirements during redox biocatalysis. At epoxidation rates of above 21 micromol per min per g cells (dry weight), the absence of limitations by O(2) and styrene and stagnating NAD(P)H regeneration rates indicated that NADH availability limited styrene epoxidation. During glucose-limited growth, oxygenase catalysis might induce regulatory stress responses, which attenuate excessive glucose catabolism and thus limit NADH regeneration. Optimizing metabolic and/or regulatory networks for efficient redox biocatalysis instead of growth (yield) is likely to be the key for maintaining high oxygenase activities in recombinant E. coli.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.