Purification and Characterization of the Acetone Carboxylase of Cupriavidus metallidurans Strain CH34

Rosier, Caroline; Leys, Natalie; Henoumont, Céline; Mergeay, Max; Wattiez, Ruddy

doi:10.1128/aem.07974-11

Cited by 15 publications

(19 citation statements)

References 20 publications

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“…Acetone carboxylases are soluble cytoplasmic enzymes present in many species of aerobic [Clark and Ensign, 1999;Rosier et al, 2012;Sluis and Ensign, 1997;Sluis et al, 2002], denitrifying [Bonnet-Smits et al, 1988;Dullius et al, 2011;Oosterkamp et al, 2015;Schühle and Heider, 2012], or anaerobic phototrophic bacteria [Birks and Kelly, 1997;Sluis et al, 2002], and even very prominent in the microaerobic gastric human pathogenic species Helicobacter pylori [Brahmachary et al, 2008;Sluis et al, 2002] ( fig. 1 ).…”

Section: Acetone Carboxylasesmentioning

confidence: 99%

“…1 ). In these species, acetone carboxylase is generally induced to high ratios of the total protein upon growth with acetone, probably to make up for the generally low specific activity of the enzyme (0.08-0.8 U/mg) [Oosterkamp et al, 2015;Rosier et al, 2012;Schühle and Heider, 2012;Sluis et al, 2002;Wöhlbrand et al, 2007].…”

Section: Acetone Carboxylasesmentioning

confidence: 99%

“…Aerobic bacteria in which acetone carboxylation was detected include the Alphaproteobacterium X. autotrophicus strain Py2 Sluis and Ensign, 1997;Sluis et al, 1996Sluis et al, , 2002, the Betaproteobacterium Cupriavidus metallidurans [Rosier et al, 2012], and the Actinobacterium Rhodococcus rhodochrous [Clark and Ensign, 1999]. In all these bacteria, acetone metabolism is initiated by a substrate-induced acetone carboxylase which is specifically produced during growth with acetone and accounts for up to 25% of total cell protein in acetone-grown cells of X. autotrophicus [Sluis et al, 2002].…”

Section: Acetone Carboxylasesmentioning

confidence: 99%

“…Acetone carboxylases have so far been purified and characterized from anaerobically grown cells of R. capsulatus [Boyd and Ensign, 2005;Boyd et al, 2004;Sluis et al, 2002] and A. aromaticum [Schühle and Heider, 2012], as well as from aerobically grown cells of X. autotrophicus, C. metallidurans , and R. rhodochrous Clark and Ensign, 1999;Rosier et al, 2012;Sluis and Ensign, 1997;Sluis et al, 2002]. All these enzymes are only able to carboxylate either acetone or butanone with bicarbonate as a cosubstrate, except for the enzyme from R. rhodochrous , which has a broader substrate range including 2-and 3-pentanone and 2-hexanone, and to utilize GTP or ITP rather than ATP [Clark and Ensign, 1999].…”

Section: Two Apparent Mechanistic Classes Of Acetone Carboxylasesmentioning

confidence: 99%

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Activation of Acetone and Other Simple Ketones in Anaerobic Bacteria

et al. 2016

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Acetone and other ketones are activated for subsequent degradation through carboxylation by many nitrate-reducing, phototrophic, and obligately aerobic bacteria. Acetone carboxylation leads to acetoacetate, which is subsequently activated to a thioester and degraded via thiolysis. Two different types of acetone carboxylases have been described, which require either 2 or 4 ATP equivalents as an energy supply for the carboxylation reaction. Both enzymes appear to combine acetone enolphosphate with carbonic phosphate to form acetoacetate. A similar but more complex enzyme is known to carboxylate the aromatic ketone acetophenone, a metabolic intermediate in anaerobic ethylbenzene metabolism in denitrifying bacteria, with simultaneous hydrolysis of 2 ATP to 2 ADP. Obligately anaerobic sulfate-reducing bacteria activate acetone to a four-carbon compound as well, but via a different process than bicarbonate- or CO₂-dependent carboxylation. The present evidence indicates that either carbon monoxide or a formyl residue is used as a cosubstrate, and that the overall ATP expenditure of this pathway is substantially lower than in the known acetone carboxylase reactions.

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Section: Acetone Carboxylasesmentioning

confidence: 99%

Section: Acetone Carboxylasesmentioning

confidence: 99%

Section: Acetone Carboxylasesmentioning

confidence: 99%

Section: Two Apparent Mechanistic Classes Of Acetone Carboxylasesmentioning

confidence: 99%

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Activation of Acetone and Other Simple Ketones in Anaerobic Bacteria

et al. 2016

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“…The activating enzyme differs from the ketone carboxylases employed by aerobic and nitratereducing bacteria, which depend on the presence of divalent cations, such as Mg 2ϩ and Mn 2ϩ (31,32). However, the acetonecarbonylating enzyme activity of D. biacutus was stimulated by NH 4 ϩ ions, similar to the acetone carboxylases of Cupriavidus metallidurans strain CH34 and Xanthobacter autotrophicus strain Py2 (13,33). Neither genomic nor proteomic analysis of acetonegrown cells of D. biacutus provided any indication of acetone carboxylases similar to those described for aerobic or nitrate-reducing acetone oxidizers (unpublished results from our labs).…”

Section: Discussionmentioning

confidence: 92%

Carbonylation as a Key Reaction in Anaerobic Acetone Activation by Desulfococcus biacutus

Acosta

Hardt

Schink

2013

Appl Environ Microbiol

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Acetone is activated by aerobic and nitrate-reducing bacteria via an ATP-dependent carboxylation reaction to form acetoacetate as the first reaction product. In the activation of acetone by sulfate-reducing bacteria, acetoacetate has not been found to be an intermediate. Here, we present evidence of a carbonylation reaction as the initial step in the activation of acetone by the strictly anaerobic sulfate reducer Desulfococcus biacutus. In cell suspension experiments, CO was found to be a far better cosubstrate for acetone activation than CO 2 . The hypothetical reaction product, acetoacetaldehyde, is extremely reactive and could not be identified as a free intermediate. However, acetoacetaldehyde dinitrophenylhydrazone was detected by mass spectrometry in cell extract experiments as a reaction product of acetone, CO, and dinitrophenylhydrazine. In a similar assay, 2-amino-4-methylpyrimidine was formed as the product of a reaction between acetoacetaldehyde and guanidine. The reaction depended on ATP as a cosubstrate. Moreover, the specific activity of aldehyde dehydrogenase (coenzyme A [CoA] acylating) tested with the putative physiological substrate was found to be 153 ؎ 36 mU mg ؊1 protein, and its activity was specifically induced in extracts of acetone-grown cells. Moreover, acetoacetyl-CoA was detected (by mass spectrometry) after the carbonylation reaction as the subsequent intermediate after acetoacetaldehyde was formed. These results together provide evidence that acetoacetaldehyde is an intermediate in the activation of acetone by sulfate-reducing bacteria.A cetone is produced by bacterial fermentations, for example, by several Clostridium species (1). It is also produced in chemistry as a solvent and as an intermediate in the synthetic chemical industry. Aerobic degradation of methyl ketones was first observed with hydrocarbon-utilizing bacteria (2). Acetone is degraded by some aerobic bacteria (3) and mammalian liver cells via oxygenase-dependent hydroxylation to acetol (4). Carboxylation of acetone to acetoacetate as a means of acetone activation was first proposed for a methanogenic enrichment culture (5). The requirement of CO 2 as a cosubstrate for acetone degradation was also observed with the nitrate reducer Thiosphaera pantotropha (6) and with Rhodobacter capsulatus and other phototrophs (7). The reaction was studied with the nitrate-reducing strain Bun N under anoxic conditions, and it was concluded that acetoacetate was formed by the ATP-dependent carboxylation of acetone (8, 9).Attempts to measure an in vitro carboxylation of acetone at that time were unsuccessful. However, exchange of radioactively labeled CO 2 with the carboxyl group of acetoacetate was catalyzed by cell extracts of strain Bun N (10). A similar CO 2 -and ATPdependent activation reaction was observed with the aerobic bacterium Xanthobacter autotrophicus strain Py2 (11). A comparison between the acetone carboxylase of strain Py2 and the carboxylase of the phototrophic bacterium Rhodobacter capsulatus showed that they are ide...

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Acetone Carboxylase

Alleman¹,

Prussia²,

Mus³

et al. 2019

Encyclopedia of Inorganic and Bioinorganic Chemistry

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Carboxylases incorporate CO 2 , or its hydrated form HCO 3 − , into organic substrates. Assimilatory carboxylases combine CO 2 to a diversity of carbon sources allowing for growth on intransigent, poorly activated organic compounds such as acetone. Acetone carboxylase catalyzes the adenosine triphosphate‐dependent carboxylation of acetone to acetoacetate. Acetone carboxylase undergoes a large conformational shift to allow phosphorylated intermediates, carboxyphosphate and phosphenolacetone to interact with the Mn bound active site. The coordinated Mn orients the intermediates for the carboncarbon bond formation producing acetoacetate. Biochemical, molecular, structural, and spectroscopic studies have shown how acetone carboxylase functions and gives clues on how this unique biotin‐independent carboxylation mechanism couples ATP hydrolysis to bicarbonate‐dependent carboxylation. This article highlights the body of biochemical, spectroscopic, and structural work leading to insights into the catalytic mechanism of acetone carboxylase.

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Purification and Characterization of the Acetone Carboxylase of Cupriavidus metallidurans Strain CH34

Cited by 15 publications

References 20 publications

Activation of Acetone and Other Simple Ketones in Anaerobic Bacteria

Activation of Acetone and Other Simple Ketones in Anaerobic Bacteria

Carbonylation as a Key Reaction in Anaerobic Acetone Activation by Desulfococcus biacutus

Acetone Carboxylase

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