ArticleApplied and environmental microbiology2013
Proline availability regulates proline-4-hydroxylase synthesis and substrate uptake in proline-hydroxylating recombinant Escherichia coli.
Article in Applied and environmental microbiology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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16 citing papers in PubMed, 38 citations in OpenAlex.
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- Modular reconstruction and optimization of the trans-4-hydroxy-L-proline synthesis pathway in Escherichia coli.Microbial cell factories · 2022Article
- Synthetic azo dye bio-decolorization by Priestia sp. RA1: process optimization and phytotoxicity assessment.Archives of microbiology · 2022Article
- Myo-Inositol in Fermented Sugar Matrix Improves Human Macrophage Function.Molecular nutrition & food research · 2022Article
- The Biosynthesis of D-1,2,4-Butanetriol From d-Arabinose With an EngineeredFrontiers in bioengineering and biotechnology · 2022Article
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- Metabolic engineering strategy for synthetizing trans-4-hydroxy-L-proline in microorganisms.Microbial cell factories · 2021Review
- Chassis engineering of Escherichia coli for trans-4-hydroxy-l-proline production.Microbial biotechnology · 2021Article
- 2-Ketoglutarate-Generated In Vitro Enzymatic Biosystem Facilitates Fe(II)/2-Ketoglutarate-Dependent Dioxygenase-Mediated C-H Bond Oxidation for (2International journal of molecular sciences · 2020Article
- Studies on the selectivity of proline hydroxylases reveal new substrates including bicycles.Bioorganic chemistry · 2020Article
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- Highly Regioselective and Stereoselective Hydroxylation of Free Amino Acids by a 2-Oxoglutarate-Dependent Dioxygenase fromACS omega · 2019Article
- Process optimization for enhancing production of cis-4-hydroxy-L-proline by engineered Escherichia coli.Microbial cell factories · 2017Article
- Identification of Trans-4-Hydroxy-L-Proline as a Compatible Solute and Its Biosynthesis and Molecular Characterization inFrontiers in microbiology · 2017Article
- Metabolic network capacity of Escherichia coli for Krebs cycle-dependent proline hydroxylation.Microbial cell factories · 2015Article
- Biosynthesis of trans-4-hydroxyproline by recombinant strains of Corynebacterium glutamicum and Escherichia coli.BMC biotechnology · 2014Article
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Authors and funding
6 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Microbial physiology plays a crucial role in whole-cell biotransformation, especially for redox reactions that depend on carbon and energy metabolism. In this study, regio- and enantio-selective proline hydroxylation with recombinant Escherichia coli expressing proline-4-hydroxylase (P4H) was investigated with respect to its interconnectivity to microbial physiology and metabolism. P4H production was found to depend on extracellular proline availability and on codon usage. Medium supplementation with proline did not alter p4h mRNA levels, indicating that P4H production depends on the availability of charged prolyl-tRNAs. Increasing the intracellular levels of soluble P4H did not result in an increase in resting cell activities above a certain threshold (depending on growth and assay temperature). Activities up to 5-fold higher were reached with permeabilized cells, confirming that host physiology and not the intracellular level of active P4H determines the achievable whole-cell proline hydroxylation activity. Metabolic flux analysis revealed that tricarboxylic acid cycle fluxes in growing biocatalytically active cells were significantly higher than proline hydroxylation rates. Remarkably, a catalysis-induced reduction of substrate uptake was observed, which correlated with reduced transcription of putA and putP, encoding proline dehydrogenase and the major proline transporter, respectively. These results provide evidence for a strong interference of catalytic activity with the regulation of proline uptake and metabolism. In terms of whole-cell biocatalyst efficiency, proline uptake and competition of P4H with proline catabolism are considered the most critical factors.
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