Biochemistry, genetics and biotechnology of glycerol utilization in <i>Pseudomonas</i> species

Poblete‐Castro, Ignacio; Wittmann, Christoph; Nikel, Pablo I.

doi:10.1111/1751-7915.13400

Cited by 87 publications

(84 citation statements)

References 189 publications

(197 reference statements)

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“…2; see also Data Set S1). Glycerol is assimilated through the processes of uptake (GlpF), phosphorylation (GlpK), and dehydrogenation (GlpD) to yield dihydroxyacetone phosphate (DHAP) (27)(28)(29). Commensurate with this, expression of all three enzymes/transporters (and of their corresponding transcripts) was strongly stimulated during growth on glycerol.…”

Section: Figmentioning

confidence: 99%

Contextual Flexibility in Pseudomonas aeruginosa Central Carbon Metabolism during Growth in Single Carbon Sources

Dolan

Kohlstedt

Trigg

et al. 2020

mBio

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Pseudomonas aeruginosa is an opportunistic human pathogen, particularly noted for causing infections in the lungs of people with cystic fibrosis (CF). Previous studies have shown that the gene expression profile of P. aeruginosa appears to converge toward a common metabolic program as the organism adapts to the CF airway environment. However, we still have only a limited understanding of how these transcriptional changes impact metabolic flux at the systems level. To address this, we analyzed the transcriptome, proteome, and fluxome of P. aeruginosa grown on glycerol or acetate. These carbon sources were chosen because they are the primary breakdown products of an airway surfactant, phosphatidylcholine, which is known to be a major carbon source for P. aeruginosa in CF airways. We show that the fluxes of carbon throughout central metabolism are radically different among carbon sources. For example, the newly recognized “EDEMP cycle” (which incorporates elements of the Entner-Doudoroff [ED] pathway, the Embden-Meyerhof-Parnas [EMP] pathway, and the pentose phosphate [PP] pathway) plays an important role in supplying NADPH during growth on glycerol. In contrast, the EDEMP cycle is attenuated during growth on acetate, and instead, NADPH is primarily supplied by the reaction catalyzed by isocitrate dehydrogenase(s). Perhaps more importantly, our proteomic and transcriptomic analyses revealed a global remodeling of gene expression during growth on the different carbon sources, with unanticipated impacts on aerobic denitrification, electron transport chain architecture, and the redox economy of the cell. Collectively, these data highlight the remarkable metabolic plasticity of P. aeruginosa; that plasticity allows the organism to seamlessly segue between different carbon sources, maximizing the energetic yield from each. IMPORTANCE Pseudomonas aeruginosa is an opportunistic human pathogen that is well known for causing infections in the airways of people with cystic fibrosis. Although it is clear that P. aeruginosa is metabolically well adapted to life in the CF lung, little is currently known about how the organism metabolizes the nutrients available in the airways. In this work, we used a combination of gene expression and isotope tracer (“fluxomic”) analyses to find out exactly where the input carbon goes during growth on two CF-relevant carbon sources, acetate and glycerol (derived from the breakdown of lung surfactant). We found that carbon is routed (“fluxed”) through very different pathways during growth on these substrates and that this is accompanied by an unexpected remodeling of the cell’s electron transfer pathways. Having access to this “blueprint” is important because the metabolism of P. aeruginosa is increasingly being recognized as a target for the development of much-needed antimicrobial agents.

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Section: Figmentioning

confidence: 99%

Contextual Flexibility in Pseudomonas aeruginosa Central Carbon Metabolism during Growth in Single Carbon Sources

Dolan

Kohlstedt

Trigg

et al. 2020

mBio

Self Cite

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show abstract

“…During growth on glycerol, there was strong induction of the glycerol uptake system at both the proteomic and transcriptomic level (Figure 2, File S1). Glycerol is assimilated through the process of uptake (GlpF), phosphorylation (GlpK) and dehydrogenation (GlpD) to yield dihydroxyacetone phosphate (DHAP) [26–28]. Commensurate with this, the expression of all three enzymes/transporters (and their corresponding transcripts) was strongly stimulated during growth on glycerol.…”

Section: Resultsmentioning

confidence: 99%

Contextual flexibility inPseudomonas aeruginosacentral carbon metabolism during growth in single carbon sources

Dolan

Kohlstedt

Trigg

et al. 2019

Preprint

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Pseudomonas aeruginosa is an opportunistic human pathogen, particularly noted for causing infections in the lungs of people with cystic fibrosis (CF). Previous studies have shown that the gene expression profile of P. aeruginosa appears to converge towards a common metabolic program as the organism adapts to the CF airway environment. However, at a systems level, we still have only a limited understanding of how these transcriptional changes impact on metabolic flux. To address this, we analysed the transcriptome, proteome and fluxome of P. aeruginosa grown on glycerol or acetate. These carbon sources were chosen because they are the primary breakdown products of airway surfactant, phosphatidylcholine, which is known to be a major carbon source for P. aeruginosa in the CF airways. We show that the flux of carbon through central metabolism is radically different on each carbon source. For example, the newly-recognised EDEMP cycle plays an important role in supplying NADPH during growth on glycerol. By contrast, the EDEMP cycle is attenuated during growth on acetate, and instead, NADPH is primarily supplied by the isocitrate dehydrogenase(s)-catalyzed reaction. Perhaps more importantly, our proteomic and transcriptomic analyses reveal a global remodelling of gene expression during growth on the different carbon sources, with unanticipated impacts on aerobic denitrification, electron transport chain architecture, and the redox economy of the cell. Collectively, these data highlight the remarkable metabolic plasticity of P. aeruginosa; a plasticity which allows the organism to seamlessly segue between different carbon sources, maximising the energetic yield from each. Importance Pseudomonas aeruginosa is an opportunistic human pathogen, well-known for causing infections in the airways of people with cystic fibrosis. Although it is clear that P. aeruginosa is metabolically well-adapted to life in the CF lung, little is currently known about how the organism metabolises the nutrients available in the airways. In this work, we use a combination of gene expression and isotope tracer (“fluxomic”) analyses to find out exactly where the input carbon goes during growth on two CF-relevant carbon sources, acetate and glycerol (derived from the breakdown of lung surfactant). We find that carbon is routed (“fluxed”) through very different pathways during growth on these substrates, and that this is accompanied by an unexpected remodelling of the cell’s electron transfer pathways. Having access to this “blueprint” is important because the metabolism of P. aeruginosa is increasingly being recognised as a target for the development of much-needed antimicrobial agents.

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“…Four P. aeruginosa strains can anaerobically grow well using the possible metabolic intermediates and analogues of glycerol. These substrates are water-soluble and easily enter into the glycolysis and gluconeogenesis pathways, which may be conducive to the rapid absorption and anaerobic metabolize by P. aeruginosa [42]. Among the possible metabolic intermediates and analogues of glycerol, only 1, 2-propylene glycol promoted four P. aeruginosa strains to produce rhamnolipids under anaerobic conditions, reducing the surface tension of anaerobic culture from 63.2 mN/m to about 31 mN/m (Fig.…”

Section: Research Perspectives To the Anaerobic Biosynthesis Mechanismentioning

confidence: 99%

“…And they found that the C 6 unit of rhamnose group in rhamnolipids product was directly condensed from the two molecules glycerol (C 3 unit) without carbon chain rearrangement [44]. The polyol can be oxidized into dihydroxyacetone (DHA), then DHA can be converted into dihydroxyacetone phosphate (DHAP) which enters glycolysis and gluconeogenesis pathways [42,43]. This may be why P. aeruginosa can metabolize glycerol and 1, 2-propylene glycol for biosynthesis the precursors of rhamnolipids, rhamnose and fatty acids.…”

Section: Research Perspectives To the Anaerobic Biosynthesis Mechanismentioning

confidence: 99%

Glycerol promoted the anaerobic production of rhamnolipids by different Pseudomonas aeruginosa strains

Zhao

Guo

Cui

2020

Preprint

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Background: Rhamnolipids is the most widely studied and applied biosurfactants. The anaerobic biosynthesis of rhamnolipids has important research and practical significance, such as meeting the in situ production of biosurfactant in anoxic environments and the foamless fermentation of biosurfactants. A few studies have reported the anaerobic biosynthesis of rhamnolipids from rare Pseudomonas aeruginosa strains. What did promote the anaerobic biosynthesis of rhamnolipids, the specificity of the rare strains or the effect of specific substrates? Here, anaerobic production of rhamnolipids by different P. aeruginosa strains was investigated using diverse substrates. The anaerobic biosynthesis mechanism of rhamnolipids were also discussed from the substrate point of view.Results: All P. aeruginosa strains anaerobically grew well using the tested substrates. But all P. aeruginosa strains anaerobically produced rhamnolipids only using substrates containing glycerol and nitrate. Fourier transform infrared (FTIR) spectra analysis confirmed the anaerobic production of rhamnolipids from all P. aeruginosa strains. All the anaerobically produced rhamnolipids decreased air-water surface tension from 72.6 mN/m to below 29.0 mN/m and emulsified crude oil with EI24 above 65%. Using crude glycerol as low-cost substrate, all P. aeruginosa strains can anaerobically grow and produce rhamnolipids to reduce the culture surface tension below 35 mN/m. The glycerol metabolic intermediate, 1, 2-propylene glycol, can also achieve the anaerobic production of rhamnolipids by all P. aeruginosa strains.Conclusions: Not the specificity of the rare P. aeruginosa strains but the effect of specific substrates promote the anaerobic biosynthesis of rhamnolipids by P. aeruginosa. Glycerol and nitrate are the excellent substrates for anaerobic production of rhamnolipids from all P. aeruginosa strains. Results indicated that glycerol metabolism involveed the anaerobic biosynthesis of rhamnolipids in P. aeruginosa. Results also showed the feasibility of using crude glycerol as low cost substrate to anaerobically biosynthesize rhamnolipids by P. aeruginosa.

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Biochemistry, genetics and biotechnology of glycerol utilization in Pseudomonas species

Cited by 87 publications

References 189 publications

Contextual Flexibility in Pseudomonas aeruginosa Central Carbon Metabolism during Growth in Single Carbon Sources

Contextual Flexibility in Pseudomonas aeruginosa Central Carbon Metabolism during Growth in Single Carbon Sources

Contextual flexibility inPseudomonas aeruginosacentral carbon metabolism during growth in single carbon sources

Glycerol promoted the anaerobic production of rhamnolipids by different Pseudomonas aeruginosa strains

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