MmcBC in<i>Pelotomaculum thermopropionicum</i>represents a novel group of prokaryotic fumarases

Shimoyama, Takefumi; Rajashekhara, Eranna; Ohmori, Daijiro; Kosaka, Tomoyuki; Watanabe, Kazuya

doi:10.1111/j.1574-6968.2007.00665.x

Cited by 20 publications

(27 citation statements)

References 22 publications

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“…The gene had a long ORF of 1371 bp (G + C content of 57.8%) that encoded a 457 amino acid protein. The predicted Mr was approximately 49.2 kDa with an isoelectric point of 4.8, similar to the 48-50 kDa Mr of most FumC proteins [7]. When the deduced amino acid sequence of FumF was searched against the NCBI and Expasy databases, FumF protein was found to share partial homology with the Class II fumarate hydratases from Bacteroides sp.…”

Section: Resultsmentioning

confidence: 96%

“…In contrast, an amino acid N-terminal extension was present only in the bacteria fumarases (Figure 1). Based on amino acid sequence homology, FumF may be a new member of the Class II fumarate hydratases [7]. …”

Section: Resultsmentioning

confidence: 99%

“…The Class I fumarases, encoded by the fumA and fumB genes, are thermolabile homodimers containing an Fe-S cluster that may catalyze the reduction of L -malate by providing fumarate as the anaerobic electron acceptor [5]. The FumA protein is stable, whereas FumB protein is unstable under aerobic conditions and functions only under anaerobic conditions [6,7]. The Class II fumarases, encoded by the fumC gene, are thermostable homotetramers with no requirement for cofactors and catalyze the interconversion of fumarate to L -malate [2].…”

Section: Introductionmentioning

confidence: 99%

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Identification and characterization of a novel fumarase gene by metagenome expression cloning from marine microorganisms

Jiang

Zhao

et al. 2010

Microb Cell Fact

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Background: Fumarase catalyzes the reversible hydration of fumarate to L-malate and is a key enzyme in the tricarboxylic acid (TCA) cycle and in amino acid metabolism. Fumarase is also used for the industrial production of L-malate from the substrate fumarate. Thermostable and high-activity fumarases from organisms that inhabit extreme environments may have great potential in industry, biotechnology, and basic research. The marine environment is highly complex and considered one of the main reservoirs of microbial diversity on the planet. However, most of the microorganisms are inaccessible in nature and are not easily cultivated in the laboratory. Metagenomic approaches provide a powerful tool to isolate and identify enzymes with novel biocatalytic activities for various biotechnological applications.

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Section: Resultsmentioning

confidence: 96%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Identification and characterization of a novel fumarase gene by metagenome expression cloning from marine microorganisms

Jiang

Zhao

et al. 2010

Microb Cell Fact

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“…Although functionally related, FHs from classes I and II exhibit low sequence identity (around 20%). Recently, a novel group of fumarate hydratases has been described in the literature as two-subunit FHs based on sequence identity to class I fumarate hydratases [11,12]. Twosubunit fumarate hydratases are oxygen-sensitive, thermostable, Fe-S cluster-containing heterodimeric proteins with two different subunits (␣ and ␤ subunit).…”

Section: Introductionmentioning

confidence: 99%

Fumarate hydratase isoforms of Leishmania major: Subcellular localization, structural and kinetic properties

Feliciano

Gupta

Dyszy

et al. 2012

International Journal of Biological Macromolecules

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Fumarate hydratases (FHs; EC 4.2.1.2) are enzymes that catalyze the reversible hydration of fumarate to S-malate. Parasitic protists that belong to the genus Leishmania and are responsible for a complex of vector-borne diseases named leishmaniases possess two genes that encode distinct putative FH enzymes. Genome sequence analysis of Leishmania major Friedlin reveals the existence of genes LmjF24.0320 and LmjF29.1960 encoding the putative enzymes LmFH-1 and LmFH-2, respectively. In the present work, the FH activity of both L. major enzymes has been confirmed. Circular dichroism studies suggest important differences in terms of secondary structure content when comparing LmFH isoforms and even larger differences when comparing them to the homologous human enzyme. CD melting experiments revealed that both LmFH isoforms are thermolabile enzymes. The catalytic efficiency under aerobic and anaerobic environments suggests that they are both highly sensitive to oxidation and damaged by oxygen. Intracellular localization studies located LmFH-1 in the mitochondrion, whereas LmFH-2 was found predominantly in the cytosol with possibly also some in glycosomes. The high degree of sequence conservation in different Leishmania species, together with the relevance of FH activity for the energy metabolism in these parasites suggest that FHs might be exploited as targets for broad-spectrum antileishmanial drugs.

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“…FH is found in two biochemically distinct forms; class I FH a thermolabile, oxygen sensitive, 4Fe-4S cluster containing enzyme and, class II FH, a stable, oxygen insensitive and ironindependent enzyme (7). Class I FH is further divided into two types, two-subunit and singlesubunit, depending on the number of genes that encode the functional enzyme (8). There is no sequence homology between these two classes of enzymes.…”

mentioning

confidence: 99%

Biochemical characterization and essentiality of fumarate hydratase

Jayaraman¹,

Suryavanshi²,

Kalale

et al. 2018

Journal of Biological Chemistry

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Plasmodium falciparum (Pf), the causative agent of malaria, has an iron-sulfur cluster-containing class I fumarate hydratase (FH) that catalyzes the interconversion of fumarate to malate, a wellknown reaction in the tricarboxylic acid cycle. In humans, the same reaction is catalyzed by class II FH that has no sequence or structural homology with the class I enzyme from Plasmodium. Fumarate is generated in large quantities in the parasite as a byproduct of AMP synthesis and is converted to malate by FH and then used in the generation of the key metabolites oxaloacetate, aspartate, and pyruvate. Previous studies have identified the FH reaction as being essential to P. falciparum, but biochemical characterizations of PfFH that may provide leads for the development of specific inhibitors are lacking. Here, we report on the kinetic characterization of purified recombinant PfFH, functional complementation of fh deficiency in Escherichia coli, and mitochondrial localization in the parasite. We found that the substrate analog mercaptosuccinic acid is a potent PfFH inhibitor, with a K i value in the nanomolar range. The fh gene could not be knocked out in Plasmodium berghei when transfectants were introduced into BALB/c mice; however, fh knockout was successful when C57BL/6 mice were used as host, suggesting that the essentiality of the fh gene to the parasite was mouse strain dependent.Plasmodium falciparum (Pf), the causative agent of the most lethal form of malaria, during its intra-erythrocytic asexual stages, derives ATP primarily from glycolysis with low contribution from mitochondrial pathways (1, 2). The bulk of pyruvate formed is converted to lactic acid with a minor amount entering the tricarboxylic acid (TCA) cycle, the flux through which is upregulated in sexual stages (2). Key intermediates that anaplerotically feed into the TCA cycle are α-ketoglutarate derived from glutamate, oxaloacetate (OAA) from phosphoenolpyruvate, and fumarate from adenosine 5΄-monophosphate (AMP) synthesis. Synthesis of AMP in the parasite is solely from inosine-5΄-monophosphate (IMP) through a pathway involving the enzymes adenylosuccinate synthetase (ADSS) and adenylosuccinate lyase (ASL). The net reaction of ADSS and ASL involves consumption of GTP and aspartate and, generation of GDP, P i and fumarate. In the rapidly dividing parasite with an AT-rich genome and high energy requirements, leading to a high demand for adenine pools, one would expect a high flux of fumarate generation. The parasite does not secrete fumarate but instead, the carbon derived from this metabolite can be traced in malate, OAA, aspartate, pyruvate (through PEP) and lactate (3). The metabolic significance of this fumarate anaplerosis is still obscure. In this context, fumarate hydratase (FH, fumarase) the key enzyme to metabolize fumarate becomes an Fumarate hydratase (fumarase, E.C. 4.2.1.2) catalyzes the reversible conversion of fumarate to malate. The stereospecific reaction involves the anti-addition of a water molecule across the carbon-carbon...

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MmcBC inPelotomaculum thermopropionicumrepresents a novel group of prokaryotic fumarases

Cited by 20 publications

References 22 publications

Identification and characterization of a novel fumarase gene by metagenome expression cloning from marine microorganisms

Identification and characterization of a novel fumarase gene by metagenome expression cloning from marine microorganisms

Fumarate hydratase isoforms of Leishmania major: Subcellular localization, structural and kinetic properties

Biochemical characterization and essentiality of fumarate hydratase

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