Pyruvate dehydrogenase complex regulator (PdhR) gene deletion boosts glucose metabolism in Escherichia coli under oxygen-limited culture conditions

Maeda, Soya; Shimizu, Kumiko; Kihira, Chie; Iwabu, Yuki; Kato, Ryuichi; Sugimoto, Makoto; Fukiya, Satoru; Wada, Masaaki; Yokota, Atsushi

doi:10.1016/j.jbiosc.2016.11.004

Cited by 18 publications

(22 citation statements)

References 16 publications

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“…We screened transcription factors which were affected by UvrY, and identified NarL and PdhR as potential transcription regulators of ahh1 based on RNAseq analysis. Since NarL and PdhR have been suggested to be negative transcription regulators in previous studies [56,57], the transcription levels of narL and pdhR are increased as expected in the RNAseq profiles of uvrY mutant (S6 file), and are supposed to repress ahh1expression consequently. However, further research to prove this hypothesis is warranted.…”

Section: Discussionmentioning

confidence: 57%

UvrY is required for the full virulence of Aeromonas dhakensis

et al. 2020

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Aeromonas dhakensis is an emerging human pathogen which causes fast and severe infections worldwide. Under the gradual pressure of lacking useful antibiotics, finding a new strategy against A. dhakensis infection is urgent. To understand its pathogenesis, we created an A. dhakensis AAK1 mini-Tn10 transposon library to study the mechanism of A. dhakensis infection. By using a Caenorhabditis elegans model, we established a screening platform for the purpose of identifying attenuated mutants. The uvrY mutant, which conferred the most attenuated toxicity toward C. elegans, was identified. The uvrY mutant was also less virulent in C2C12 fibroblast and mice models, in line with in vitro results. To further elucidate the mechanism of UvrY in controlling the toxicity in A. dhakensis, we conducted a transcriptomic analysis. The RNAseq results showed that the expression of a unique hemolysin ahh1 and other virulence factors were regulated by UvrY. Complementation of Ahh1, one of the most important virulence factors, rescued the pore-formation phenotype of uvrY mutant in C. elegans; however, complementation of ahh1 endogenous promoter-driven ahh1 could not produce Ahh1 and rescue the virulence in the uvrY mutant. These findings suggest that UvrY is required for the expression of Ahh1 in A. dhakensis. Taken together, our results suggested that UvrY controls several different virulence factors and is required for the full virulence of A. dhakensis. The two-component regulator UvrY therefore a potential therapeutic target which is worthy of further study.

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Section: Discussionmentioning

confidence: 57%

UvrY is required for the full virulence of Aeromonas dhakensis

et al. 2020

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“…To further confirm that RyhB deletion resulted in a metabolically hyperactive state, intracellular NAD + /NADH ratios were measured (Maeda et al, 2017 ). We found Δ ryhB mutant showed reduced NAD + /NADH ratios in both exponential (2.76-fold decrease) and stationary phase (3.66-fold decrease) cultures (Table 2 ) when compared with the parent strain.…”

Section: Resultsmentioning

confidence: 99%

“…Intracellular NAD + and NADH concentrations were measured according to methods described previously (Vilcheze et al, 2005 ; Maeda et al, 2017 ). For NAD + and NADH extraction, bacteria were cultured to the designed timepoints and harvested by centrifugation at 14,000 rpm at 4°C for 3 min.…”

Section: Methodsmentioning

confidence: 99%

Small Non-coding RNA RyhB Mediates Persistence to Multiple Antibiotics and Stresses in Uropathogenic Escherichia coli by Reducing Cellular Metabolism

Zhang

Liu

et al. 2018

Front. Microbiol.

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As dormant phenotypic variants of bacteria, persisters account for many chronic infections affecting human health. Despite numerous studies, the role of small non-coding RNA (sRNA) in bacterial persistence has not been reported. To investigate the role of Hfq-interacting sRNA in persistence, we constructed the deletion mutants of 20 Hfq-interacting sRNAs (RyhB, GcvB, MgrR, RybB, MicF, SgrS, RprA, DicF, SsrS, FnrS, GadY, DsrA, OmrB, ArcZ, RyeB, RydC, OmrA, MicA, MicC, and ChiX) to assess their persistence capacity in uropathogenic Escherichia coli strain UTI89 and identified a new sRNA RyhB being involved in persister formation. The ryhB-knockout mutant had significant defect in persistence to a diverse range of antibiotics (levofloxacin, cefotaxime, gentamicin) and stresses (hyperosmosis, acid, and heat) in both exponential phase and stationary phase. In addition, the effect of RyhB on persistence was synergistic with ppGpp and Fur protein. RNA-Seq analysis indicated that the ryhB-knockout mutant had a hyperactive metabolic state compared with the parent strain. Interestingly, increased adenosine triphosphate (ATP) levels and altered NAD+/NADH ratios were observed in the ryhB-knockout mutant. Our findings represent a new level of persistence regulation via sRNA and may provide novel therapeutic targets for interventions.

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“…The pyruvate dehydrogenase complex regulator (PdhR) is a transcriptional regulator that negatively controls the formation of the pyruvate dehydrogenase complex (PDHc), NADH dehydrogenase (NDH)-2 and cytochrome bo3 oxidase in E. coli. Under oxygen-limited conditions, it has been reported that pdhR-deficient mutants grow and consume glucose more efficiently than the WT strain, with enhanced respiration because of the increased activities of PDHc and NDH-2 [28].…”

Section: Performance Of the Mfc With Single-gene Knockout Mutantsmentioning

confidence: 99%

Promoted performance of microbial fuel cells using Escherichia coli cells with multiple-knockout of central metabolism genes

Ojima

Kawaguchi

Fukui

et al. 2019

Bioprocess Biosyst Eng

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The effect of central metabolic activity of Escherichia coli cells acting as biocatalysts on the performance of microbial fuel cells (MFCs) was studied with glucose used as the energy source. Milliliter-scale two-chambered MFCs were used with 2-hydroxy-1,4-naphthoquinone (HNQ) as an electron mediator. Among the single-gene deletions examined, frdA, pdhR, ldhA, and adhE increased the average power output of the constructed MFC. Next, multiple-gene knockout mutants were constructed using P1 transduction. The Δ5 (ΔfrdAΔpdhRΔldhAΔadhEΔpta) strain showed the highest ave. power output (1.82 mW) and coulombic efficiency (21.3%). Our results show that the combination of multiple-gene knockout in E. coli cells leads to the development of an excellent catalyst for MFCs. Finally, preventing a decrease in the pH of the anodic solution was a key factor for improving the power output of the Δ5 strain, and a maximum ave. power output of 2.21 mW was achieved with 5% NaHCO3 in the buffer. The ave. power density of the constructed MFC was 0.27 mW/cm 3 , which is comparable to an enzymatic fuel cell of a Milliliter-scale using glucose dehydrogenase.

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Pyruvate dehydrogenase complex regulator (PdhR) gene deletion boosts glucose metabolism in Escherichia coli under oxygen-limited culture conditions

Cited by 18 publications

References 16 publications

UvrY is required for the full virulence of Aeromonas dhakensis

UvrY is required for the full virulence of Aeromonas dhakensis

Small Non-coding RNA RyhB Mediates Persistence to Multiple Antibiotics and Stresses in Uropathogenic Escherichia coli by Reducing Cellular Metabolism

Promoted performance of microbial fuel cells using Escherichia coli cells with multiple-knockout of central metabolism genes

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