scholarly journals Verrucomicrobial methanotrophs grow on diverse C3 compounds and use a homolog of particulate methane monooxygenase to oxidize acetone

2021 ◽  
Author(s):  
Samuel Imisi Awala ◽  
Joo-Han Gwak ◽  
Yong-Man Kim ◽  
So-Jeong Kim ◽  
Andrea Strazzulli ◽  
...  
Keyword(s):  
Energy Source ◽  
Methane Monooxygenase ◽  
Ozone Production ◽  
Resting Cells ◽  
Growth Substrate ◽  
Particulate Methane Monooxygenase ◽  
Photochemical Pollution ◽  
Geothermal Environments ◽  
Physiological Tests ◽  
Sole Energy

AbstractShort-chain alkanes (SCA; C2-C4) emitted from geological sources contribute to photochemical pollution and ozone production in the atmosphere. Microorganisms that oxidize SCA and thereby mitigate their release from geothermal environments have rarely been studied. In this study, propane-oxidizing cultures could not be grown from acidic geothermal samples by enrichment on propane alone, but instead required methane addition, indicating that propane was co-oxidized by methanotrophs. “Methylacidiphilum” isolates from these enrichments did not grow on propane as a sole energy source but unexpectedly did grow on C3 compounds such as 2-propanol, acetone, and acetol. A gene cluster encoding the pathway of 2-propanol oxidation to pyruvate via acetol was upregulated during growth on 2-propanol. Surprisingly, this cluster included one of three genomic operons (pmoCAB3) encoding particulate methane monooxygenase (PMO), and several physiological tests indicated that the encoded PMO3 enzyme mediates the oxidation of acetone to acetol. Acetone-grown resting cells oxidized acetone and butanone but not methane or propane, implicating a strict substrate specificity of PMO3 to ketones instead of alkanes. Another PMO-encoding operon, pmoCAB2, was induced only in methane-grown cells, and the encoded PMO2 could be responsible for co-metabolic oxidation of propane to 2-propanol. In nature, propane probably serves primarily as a supplemental growth substrate for these bacteria when growing on methane.

scholarly journals Methanobactin from Methylocystis sp. Strain SB2 Affects Gene Expression and Methane Monooxygenase Activity in Methylosinus trichosporium OB3b

2015 ◽  
Vol 81 (7) ◽  
pp. 2466-2473 ◽  
Author(s):  
Muhammad Farhan Ul-Haque ◽  
Bhagyalakshmi Kalidass ◽  
Alexey Vorobev ◽  
Bipin S. Baral ◽  
Alan A. DiSpirito ◽  
...  
Keyword(s):  
Gene Expression ◽  
Methane Monooxygenase ◽  
Particulate Methane Monooxygenase ◽  
Methylosinus Trichosporium Ob3b ◽  
Methylosinus Trichosporium ◽  
Monooxygenase Activity ◽  
Content Type ◽  
Soluble Methane Monooxygenase ◽  
A Subunit ◽  
Membrane Bound

ABSTRACTMethanotrophs can express a cytoplasmic (soluble) methane monooxygenase (sMMO) or membrane-bound (particulate) methane monooxygenase (pMMO). Expression of these MMOs is strongly regulated by the availability of copper. Many methanotrophs have been found to synthesize a novel compound, methanobactin (Mb), that is responsible for the uptake of copper, and methanobactin produced byMethylosinus trichosporiumOB3b plays a key role in controlling expression of MMO genes in this strain. As all known forms of methanobactin are structurally similar, it was hypothesized that methanobactin from one methanotroph may alter gene expression in another. WhenMethylosinus trichosporiumOB3b was grown in the presence of 1 μM CuCl2, expression ofmmoX, encoding a subunit of the hydroxylase component of sMMO, was very low.mmoXexpression increased, however, when methanobactin fromMethylocystissp. strain SB2 (SB2-Mb) was added, as did whole-cell sMMO activity, but there was no significant change in the amount of copper associated withM. trichosporiumOB3b. IfM. trichosporiumOB3b was grown in the absence of CuCl2, themmoXexpression level was high but decreased by several orders of magnitude if copper prebound to SB2-Mb (Cu-SB2-Mb) was added, and biomass-associated copper was increased. Exposure ofMethylosinus trichosporiumOB3b to SB2-Mb had no effect on expression ofmbnA, encoding the polypeptide precursor of methanobactin in either the presence or absence of CuCl2.mbnAexpression, however, was reduced when Cu-SB2-Mb was added in both the absence and presence of CuCl2. These data suggest that methanobactin acts as a general signaling molecule in methanotrophs and that methanobactin “piracy” may be commonplace.

scholarly journals Towards a unified understanding of the copper sites in particulate methane monooxygenase: An X-ray absorption spectroscopic investigation

2021 ◽  
Author(s):  
George Cutsail III ◽  
Matthew O Ross ◽  
Amy C Rosenzweig ◽  
Serena DeBeer
Keyword(s):  
Greenhouse Gas ◽  
Spectroscopic Investigation ◽  
Liquid Fuel ◽  
Methane Monooxygenase ◽  
Enzymatic Conversion ◽  
Particulate Methane Monooxygenase ◽  
X Ray ◽  
Mild Conditions ◽  
Copper Sites ◽  
X Ray Absorption

The enzymatic conversion of the greenhouse gas, methane, to a liquid fuel, methanol, is performed by methane monooxygenases (MMOs) under mild conditions. The copper stoichiometry of particulate MMO (pMMO) has...

scholarly journals Polarized ATR-FTIR Spectroscopy of the Membrane-Embedded Domains of the Particulate Methane Monooxygenase†

Biochemistry ◽  
2004 ◽  
Vol 43 (42) ◽  
pp. 13283-13292 ◽  
Author(s):  
Madhuri S. Vinchurkar ◽  
Kelvin H.-C. Chen ◽  
Steve S.-F. Yu ◽  
Shan-Jen Kuo ◽  
Hui-Chi Chiu ◽  
...  
Keyword(s):  
Ftir Spectroscopy ◽  
Methane Monooxygenase ◽  
Particulate Methane Monooxygenase

scholarly journals Expression of Individual Copies ofMethylococcus capsulatus Bath Particulate Methane Monooxygenase Genes

2001 ◽  
Vol 183 (5) ◽  
pp. 1810-1812 ◽  
Author(s):  
Sergei Stolyar ◽  
Marion Franke ◽  
Mary E. Lidstrom
Keyword(s):  
Methane Monooxygenase ◽  
Gene Clusters ◽  
Gene Fusions ◽  
Methylococcus Capsulatus ◽  
Chromosomal Gene ◽  
Particulate Methane Monooxygenase ◽  
Relative Expression

ABSTRACT The expression of the two gene clusters encoding the particulate methane monooxygenase (pMMO) in Methylococcus capsulatusBath was assessed by analysis of transcripts and by use of chromosomal gene fusions. The results suggest that the two clusters are functionally redundant but that relative expression alters depending on the copper levels available for growth.

Detection of Methanotrophs in Groundwater by PCR

1999 ◽  
Vol 65 (2) ◽  
pp. 648-651 ◽  
Author(s):  
Y. S. Cheng ◽  
J. L. Halsey ◽  
K. A. Fode ◽  
C. C. Remsen ◽  
M. L. P. Collins
Keyword(s):  
Reverse Transcriptase ◽  
Environmental Samples ◽  
Methane Monooxygenase ◽  
Partial Sequence ◽  
Methanotrophic Bacteria ◽  
Particulate Methane Monooxygenase ◽  
Significant Potential ◽  
Pcr Product ◽  
Specific Amplification ◽  
Template Dna

ABSTRACT Methanotrophic bacteria have significant potential for bioremediation, which would require methods for monitoring the presence and activity of these organisms in environmental samples. In this study, PCR was used to detect methanotrophic bacteria. Primers were designed on the basis of a partial sequence of pmoA, which encodes one of the proteins of the particulate methane monooxygenase. Specific amplification of a portion of pmoA was obtained with template DNA isolated from lab strains of methanotrophs. ApmoA product was also obtained by using DNA from groundwater. The identity of the PCR product was confirmed by sequencing or by amplification with a nested primer. Reverse transcriptase PCR detected pmoA mRNA.

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