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Core metabolism shifts of methanol vs. methane growth in the methanotroph Methylomicrobium buryatense 5GB1

ABSTRACT: Core metabolism shifts of methanol vs. methane growth in the methanotroph Methylomicrobium buryatense 5GB1

PROVIDER: PRJNA433951 | ENA |

REPOSITORIES: ENA

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Core metabolism shifts of methanol vs. methane growth in the methanotroph Methylomicrobium buryatense 5GB1

Project description:Methylomicrobium buryatense 5GB1 is an obligate methylotroph, which grows on methane or methanol with similar growth rates. Core metabolic pathways are similar on both substrates, but recent studies of methane metabolism suggest that growth on methanol might have significant differences from growth on methane. In this study, both a targeted metabolomics approach as well as a 13C tracer approach have been taken to understand core carbon metabolism in M. buryatense 5GB1 during methanol growth, to determine whether such differences occur. Targeted metabolomics analyses were performed on both methane and methanol cultures to identify metabolic nodes with altered fluxes. Several key metabolites showed significant differences in pool size. Noticeably, 2-keto-3-deoxy-6-phosphogluconate (KDPG) showed much larger pools under methanol culture, suggesting the Entner-Doudoroff (ED) pathway was more active. Intermediates in other parts of metabolism also showed differences in pool sizes under methanol growth. A systematic shift of active core metabolism is proposed to explain the changes. In order to distinguish flux partition differences at the C3-C4 node, 13C tracer analysis was also applied to methanol-grown cultures. Using the experimental results as constraints, we applied flux balance analysis to determine the metabolic flux phenotype of M. buryatense 5GB1 growing on methanol. The resulting new insights into core metabolism of this methanotroph provide an improved basis for future strain design.

2019-04-17 | GSE110541 | GEO

OXYGEN-LIMITED METABOLISM IN THE METHANOTROPH METHYLOMICROBIUM BURYATENSE 5GB1C

Project description:The bacteria that grow on methane aerobically (methanotrophs) support populations of non-methanotrophs in the natural environment by excreting methane-derived carbon. One group of excreted compounds are short-chain organic acids, generated in highest abundance when cultures are grown under O2-starvation. We examined this O2-starvation condition in the methanotroph Methylomicrobium buryatense 5GB1C . Under prolonged O2-starvation in a closed vial, this methanotroph increases the amount of acetate excreted about 10-fold, but the formate, lactate, and succinate excreted do not respond to this culture condition. In bioreactor cultures, the amount of each excreted product is similar across a range of growth rates and limiting substrates, including O2-limitation. A set of mutants were generated in genes predicted to be involved in generating or regulating excretion of these compounds and tested for growth defects, and changes in excretion products. The phenotypes and associated metabolic flux modeling suggested that in M. buryatense 5GB1C, formate and acetate are excreted in response to redox imbalance, and the resulting metabolic state represents a combination of fermentation and respiration metabolism.

2017-08-24 | GSE101981 | GEO

Core Metabolism Shifts during Growth on Methanol versus Methane in the Methanotroph Methylomicrobium buryatense 5GB1.

Project description: Not available

| S-EPMC6456754 | biostudies-literature

Methylotuvimicrobium buryatense

Project description:OXYGEN-LIMITED METABOLISM IN THE METHANOTROPH METHYLOMICROBIUM BURYATENSE 5GB1C

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XoxF Acts as the Predominant Methanol Dehydrogenase in the Type I Methanotroph Methylomicrobium buryatense.

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Methylotuvimicrobium buryatense

Project description:Methylomicrobium buryatense overview

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A mutagenic screen identifies a TonB-dependent receptor required for the lanthanide metal switch in the Type I methanotroph “Methylotuvimicrobium buryatense” 5GB1C

Project description:Several of the metabolic enzymes in methanotrophic bacteria rely on metals for both their expression and their catalysis. The MxaFI methanol dehydrogenase enzyme complex uses calcium as a cofactor to oxidize methanol, while the alternative methanol dehydrogenase XoxF uses lanthanide metals such as lanthanum and cerium for the same function. Lanthanide metals, abundant in the earth’s crust and widely used in electronic devices, strongly repress the transcription of mxaF yet activate the transcription of xoxF. This phenomenon of mxaF repression and xoxF activation in the presence of lanthanides is called the “lanthanide switch.” To better understand components of the lanthanide switch in the Type I gammaproteobacterial methanotroph “Methylotuvimicrobium buryatense” 5GB1C, we designed a mutagenesis system and selected for mutants unable to repress the mxaF promoter in the presence of lanthanum. Whole genome resequencing for multiple lanthanide switch mutants identified several unique point mutations in a single gene encoding a TonB-dependent receptor, which we have named LanA. While the LanA TonB-dependent receptor is absolutely required for the lanthanide switch, it does not affect lanthanum uptake by the bacterium. Deletion of a separate lanthanide-repressible gene encoding a TonB protein had no effect on the lanthanide switch or on lanthanum uptake. The discovery of this novel component of the lanthanide regulatory system highlights the complexity of this circuit and suggests further components are likely involved.

2019-05-21 | GSE125909 | GEO

Structure and function of the lanthanide-dependent methanol dehydrogenase XoxF from the methanotroph Methylomicrobium buryatense 5GB1C.

Project description: Not available

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Genetic tools for the industrially promising methanotroph Methylomicrobium buryatense.

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Oxygen-limited metabolism in the methanotroph Methylomicrobium buryatense 5GB1C.

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