GO:0018729 propionate CoA-transferase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0018729 propionate CoA-transferase activity catalyzes the reversible transfer of coenzyme A from acetyl-CoA to propanoate, yielding acetate and propanoyl-CoA.
• The enzyme belongs to the class of CoA-transferases (EC 2.8.3.-) and is best characterized in Clostridium propionicum and Ralstonia eutropha.
• A conserved glutamate residue (Glu324 in the C. propionicum enzyme) acts as the catalytic nucleophile during the ping-pong mechanism.
• Propionate CoA-transferase is a key enzyme in the fermentation of lactate and other substrates to propionate and in the biosynthesis of lactate esters.
• The enzyme is also found in anaerobic mitochondria of parasites such as Fasciola hepatica and Ascaris suum, where it participates in unusual energy metabolism.
• Understanding GO:0018729 is relevant for metabolic engineering, production of bioplastics, and drug development against anaerobic pathogens.
Description
Propionate CoA-transferase activity (GO:0018729) is a molecular function that enables the reversible transfer of a coenzyme A (CoA) moiety from acetyl-CoA to propanoate, producing acetate and propanoyl-CoA. This reaction is central to the metabolism of propionate and short-chain fatty acids in various microorganisms and parasites. The enzyme was first purified and characterized from Clostridium propionicum, where it plays a crucial role in the fermentation pathway that converts lactate to propionate. Later studies identified a homologous enzyme in Ralstonia eutropha H16, highlighting its broader distribution and potential biotechnological applications. Researchers study this activity to understand microbial fermentation, to engineer metabolic pathways for the production of value-added chemicals such as lactate esters, and to target parasitic diseases that rely on unusual CoA-transferases for energy generation.
propionate CoA-transferase activity At A Glance
| GO ID | GO:0018729 |
|---|---|
| GO term | propionate CoA-transferase activity |
| Ontology | molecular_function |
| Synonym | acetyl-CoA:propanoate CoA-transferase activity; propionate-CoA:lactoyl-CoA transferase activity; propionate coenzyme A-transferase activity; propionyl CoA:acetate CoA transferase activity; propionyl-CoA transferase activity |
| Definition | Catalysis of the reaction: acetyl-CoA + propanoate = acetate + propanoyl-CoA. |
| Major function | Transfer of CoA from acetyl-CoA to propanoate, yielding acetate and propanoyl-CoA; involved in propionate metabolism and fermentation. |
| EC number | 2.8.3.- (CoA-transferases) |
| Catalytic residue | Glutamate 324 in Clostridium propionicum propionate CoA-transferase |
| Representative organisms | Clostridium propionicum, Ralstonia eutropha, Ascaris suum, Fasciola hepatica |
What Is GO:0018729?
According to the Gene Ontology, propionate CoA-transferase activity (GO:0018729) is defined as the catalysis of the reaction: acetyl-CoA + propanoate = acetate + propanoyl-CoA. In other words, the enzyme transfers the CoA group from acetyl-CoA to propanoate, forming propanoyl-CoA and acetate. This is a reversible reaction that belongs to the class of CoA-transferases (EC 2.8.3.-). The activity is synonymous with acetyl-CoA:propanoate CoA-transferase, propionate-CoA:lactoyl-CoA transferase, propionate coenzyme A-transferase, propionyl CoA:acetate CoA transferase, and propionyl-CoA transferase.
Why Is propionate CoA-transferase activity Important in Cell Biology?
Propionate CoA-transferase activity is important because it links propionate metabolism to central carbon and energy metabolism. In anaerobic bacteria such as Clostridium propionicum, it is part of the pathway that converts lactate to propionate, a fermentation product that helps maintain redox balance. In the biotechnological context, the enzyme from Ralstonia eutropha has been explored for the production of lactate esters, which are green solvents. In parasites, related CoA-transferases are involved in unusual mitochondrial energy metabolism, making them potential drug targets. Moreover, the enzyme's ability to catalyze reversible CoA transfer makes it a useful tool in metabolic engineering for the production of various CoA derivatives.
• Key enzyme in propionate fermentation and lactate metabolism in Clostridium propionicum.
• Involved in the production of lactate esters, which are biodegradable solvents.
• Found in the anaerobic mitochondria of parasites like Fasciola hepatica, where it participates in energy generation.
• Contributes to the metabolism of short-chain fatty acids in Ascaris suum muscle mitochondria.
• Potential target for antiparasitic drugs due to its essential role in parasite energy metabolism.
• Useful for metabolic engineering of pathways for bioplastic and chemical production.
• Provides a model for studying CoA-transferase mechanism and evolution.
• Relevant to understanding propionate metabolism in human gut microbiota and its effects on host health.
What Happens During propionate CoA-transferase activity?
Substrate binding and initial complex formation
In simple terms: The enzyme grabs acetyl-CoA and propanoate to start the reaction.
The reaction begins with the binding of acetyl-CoA and propanoate to the active site of propionate CoA-transferase. The enzyme forms a ternary complex with both substrates, positioning the CoA moiety of acetyl-CoA near the carboxyl group of propanoate. This step is reversible and does not involve any cofactors other than the substrates themselves.
Formation of enzyme-CoA intermediate
In simple terms: The enzyme temporarily holds onto the CoA group, forming a covalent intermediate.
A conserved glutamate residue (Glu324 in Clostridium propionicum) acts as a nucleophile and attacks the thioester bond of acetyl-CoA, forming a covalent enzyme-CoA thioester intermediate and releasing acetate. This ping-pong mechanism is characteristic of CoA-transferases.
Transfer of CoA to propanoate
In simple terms: The CoA group is handed over to propanoate, making propanoyl-CoA.
The enzyme-bound CoA is then transferred to propanoate, yielding propanoyl-CoA and regenerating the free enzyme. This second half-reaction completes the catalytic cycle. The overall reaction is reversible, allowing the enzyme to also catalyze the reverse transfer from propanoyl-CoA to acetate.
Role in fermentation pathways
In simple terms: This enzyme helps bacteria turn lactate into propionate for energy balance.
In Clostridium propionicum, propionate CoA-transferase is part of the acrylate pathway, which converts lactate to propionate. The enzyme catalyzes the final step, transferring CoA from acetyl-CoA to propanoate to produce propanoyl-CoA, which is then reduced to propionate. This pathway allows the bacteria to generate ATP and maintain redox balance during fermentation.
Occurrence in parasites and unusual mitochondria
In simple terms: Some parasites use a similar enzyme in their special mitochondria to make energy.
In the anaerobic mitochondria of Fasciola hepatica, an acetate:succinate CoA-transferase (ASCT) is present, which is related to propionate CoA-transferase. This enzyme allows the parasite to produce acetate and ATP under anaerobic conditions. Similarly, Ascaris suum muscle mitochondria contain acyl-CoA transferases that participate in fermentation pathways.
Key Genes Involved in GO:0018729 propionate CoA-transferase activity
The following genes and proteins are directly or functionally associated with propionate CoA-transferase activity (GO:0018729) based on published literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| pct (Clostridium propionicum) | Encodes propionate CoA-transferase; catalyzes CoA transfer from acetyl-CoA to propanoate | Model enzyme for mechanism and active-site studies; Glu324 identified as catalytic residue |
| pct (Ralstonia eutropha H16) | Propionate CoA-transferase involved in propionate metabolism | Biotechnological applications; characterized for substrate specificity |
| ASCT (Fasciola hepatica) | Acetate:succinate CoA-transferase in anaerobic mitochondria | Potential drug target; related to propionate CoA-transferase |
| Acyl-CoA transferase (Ascaris suum) | Mitochondrial acyl-CoA transferase | Studied for anaerobic energy metabolism |
| lactate ester biosynthesis genes | Pathway for lactate ester production | Engineered using propionate CoA-transferase for green solvent production |
| carnitine palmitoyltransferase (CPT) | Fatty acid oxidation; inhibited by lactate | Indirectly related to CoA transfer; studied in cardiac muscle |
| malonyl-CoA-sensitive carnitine acyltransferase | Fatty acid metabolism in sarcoplasmic reticulum | Related CoA transfer activity in heart |
| propionate CoA-transferase homologs | Various CoA transfer reactions | Comparative genomics and evolution |
| acetyl-CoA hydrolase/transferase | Acetyl-CoA metabolism | Potential alternative routes |
| succinyl-CoA:acetate CoA-transferase | Similar CoA transfer in anaerobic microbes | Functional analog |
| butyryl-CoA:acetate CoA-transferase | Butyrate metabolism | Related enzyme family |
| 4-hydroxybutyrate CoA-transferase | GABA metabolism | Related CoA transferase |
| glutaconate CoA-transferase | Glutaconate fermentation | Model for CoA transferase mechanism |
| oxalate CoA-transferase | Oxalate metabolism | Related enzyme |
| malonate CoA-transferase | Malonate metabolism | Related enzyme |
| methylmalonyl-CoA mutase | Propionate metabolism | Downstream of propionyl-CoA |
| propionyl-CoA carboxylase | Propionate catabolism | Uses propanoyl-CoA produced by GO:0018729 |
| methylcitrate synthase | Propionate detoxification | Links to propionate metabolism |
How Is propionate CoA-transferase activity Regulated?
The regulation of propionate CoA-transferase activity is not extensively studied at the transcriptional level. In Clostridium propionicum, the pct gene is likely regulated in response to substrate availability and fermentation conditions, but specific regulators have not been identified. In Ralstonia eutropha, the enzyme may be part of a broader metabolic network controlled by global regulators of carbon metabolism. In parasites, the expression of related CoA-transferases may be developmentally regulated to adapt to anaerobic environments. Post-translational modifications have not been reported for this enzyme. Further research is needed to elucidate regulatory mechanisms.
propionate CoA-transferase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| pct (Clostridium propionicum) | Bacterial fermentation; not directly linked to human disease | Knockout in C. propionicum to study fermentation; heterologous expression in E. coli |
| ASCT (Fasciola hepatica) | Parasitic infection (fascioliasis) | RNAi knockdown in F. hepatica; enzyme inhibition assays |
| Acyl-CoA transferase (Ascaris suum) | Ascariasis; anaerobic metabolism | Gene knockout in C. elegans as a model |
| CPT1 (carnitine palmitoyltransferase) | Cardiac fatty acid oxidation disorders | Knockout mice; cardiac cell models |
| Propionyl-CoA carboxylase | Propionic acidemia | Patient-derived fibroblasts; mouse models |
Parasitic infections
Propionate CoA-transferase and related CoA-transferases are essential for the anaerobic energy metabolism of parasites such as Fasciola hepatica and Ascaris suum. Inhibition of these enzymes could disrupt ATP production, making them potential targets for antiparasitic drugs.
Metabolic disorders
Propionate metabolism is linked to disorders such as propionic acidemia, caused by deficiency of propionyl-CoA carboxylase. Although propionate CoA-transferase is not directly mutated in this disease, it participates in the pathway that generates propanoyl-CoA, the substrate for propionyl-CoA carboxylase. Understanding its role may provide insights into disease mechanisms.
Cardiac metabolism
Lactate and oxfenicine inhibit carnitine palmitoyl-CoA transferase activity and fatty acid oxidation in cardiac muscle, indicating a link between CoA transferases and cardiac energy metabolism. While not directly propionate CoA-transferase, these findings highlight the importance of CoA transfer reactions in heart function.
From propionate CoA-transferase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| What is the catalytic mechanism of propionate CoA-transferase? | Site-directed mutagenesis of Glu324 in C. propionicum pct; kinetic studies |
| How does the enzyme contribute to fermentation? | Knockout of pct in Clostridium propionicum; metabolite analysis |
| Can the enzyme be used for lactate ester production? | Overexpression of pct in engineered E. coli or yeast; bioconversion |
| Is the enzyme essential in parasites? | RNAi knockdown in Fasciola hepatica; enzyme activity assays |
| What is the substrate specificity? | Purified enzyme from Ralstonia eutropha; kinetic assays with various CoA donors and acceptors |
| How is the enzyme regulated? | Transcriptional reporter fusions; promoter analysis in C. propionicum |
How to Study the propionate CoA-transferase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Spectrophotometric enzyme assay | CoA transfer activity | Kinetic characterization of purified enzyme |
| Site-directed mutagenesis | Role of specific residues | Identification of catalytic nucleophile |
| Heterologous expression | Protein production | Purification for structural studies |
| GC-MS metabolite analysis | Substrate and product concentrations | Fermentation pathway engineering |
| RNAi knockdown | Gene function in parasites | Target validation in Fasciola hepatica |
| 13C flux analysis | Metabolic flux distribution | Metabolic engineering |
| Enzyme inhibition assays | Inhibitor potency | Drug discovery against parasites |
| Structural biology (X-ray crystallography) | Three-dimensional structure | Mechanistic insights |
Enzyme activity assays
Propionate CoA-transferase activity is typically measured spectrophotometrically by coupling the formation of propanoyl-CoA to a downstream reaction, such as citrate synthase, or by using Ellman's reagent to detect free CoA. These assays allow determination of kinetic parameters and substrate specificity.
Site-directed mutagenesis
To identify catalytic residues, site-directed mutagenesis is used. For example, mutation of Glu324 in Clostridium propionicum propionate CoA-transferase to alanine or glutamine abolishes activity, confirming its essential role.
Heterologous expression and purification
The pct gene can be cloned and overexpressed in E. coli, followed by affinity purification. This provides sufficient protein for structural and biochemical studies.
Metabolic engineering and flux analysis
In metabolic engineering, the enzyme is expressed in heterologous hosts to redirect carbon flux toward desired products such as lactate esters. Flux analysis using 13C-labeled substrates can quantify the contribution of the enzyme to product formation.
How CRISPR Can Be Used to Study GO:0018729 propionate CoA-transferase activity
Knockout
CRISPR-Cas9 knockout of the pct gene in Clostridium propionicum or related bacteria can be used to study the role of propionate CoA-transferase in fermentation and propionate production. Knockout strains may exhibit altered metabolite profiles, which can be analyzed by GC-MS.
Point Mutation
CRISPR-mediated point mutations can be introduced into the pct gene to alter specific amino acids, such as Glu324, to test their role in catalysis. This approach allows precise dissection of the catalytic mechanism without affecting other parts of the enzyme.
Knock-in
Knock-in of a tagged version of the pct gene (e.g., His-tag or FLAG-tag) can facilitate protein purification and localization studies. This is useful for determining the subcellular localization of the enzyme in parasites or bacteria.
Overexpression
CRISPR activation (CRISPRa) or plasmid-based overexpression can be used to increase propionate CoA-transferase levels in heterologous hosts for metabolic engineering applications, such as the production of lactate esters or other CoA derivatives.
How EDITGENE Supports propionate CoA-transferase activity Research
Researchers studying propionate CoA-transferase activity-related genes often need to determine whether a candidate gene is causally involved in a specific metabolic pathway or disease phenotype. EDITGENE provides a comprehensive suite of CRISPR-based services to enable precise genome editing in a wide range of cell models, from bacteria to human cells.
Contact EDITGENE today to design your custom CRISPR model for propionate CoA-transferase activity research.
Frequently Asked Questions About propionate CoA-transferase activity
What is propionate CoA-transferase activity?
Propionate CoA-transferase activity (GO:0018729) is a molecular function that catalyzes the reversible transfer of coenzyme A from acetyl-CoA to propanoate, producing acetate and propanoyl-CoA.
What genes are involved in propionate CoA-transferase activity?
The primary gene is pct, encoding propionate CoA-transferase, found in Clostridium propionicum and Ralstonia eutropha. Related genes include ASCT in Fasciola hepatica and acyl-CoA transferases in Ascaris suum.
What is the reaction catalyzed by propionate CoA-transferase?
The enzyme catalyzes: acetyl-CoA + propanoate = acetate + propanoyl-CoA. This is a reversible CoA transfer reaction.
Which organisms have propionate CoA-transferase?
It is found in anaerobic bacteria such as Clostridium propionicum and Ralstonia eutropha, as well as in parasites like Fasciola hepatica and Ascaris suum.
What is the role of glutamate 324 in propionate CoA-transferase?
Glutamate 324 in Clostridium propionicum propionate CoA-transferase acts as the catalytic nucleophile, forming a covalent enzyme-CoA intermediate during catalysis.
How is propionate CoA-transferase used in biotechnology?
It is used in metabolic engineering for the production of lactate esters and other CoA derivatives, and for converting lactate to propionate in fermentation processes.
Is propionate CoA-transferase a drug target?
Related CoA-transferases in parasites are considered potential drug targets because they are essential for anaerobic energy metabolism. Inhibition could kill the parasite.
What diseases are associated with propionate CoA-transferase?
It is not directly linked to a human genetic disease, but it participates in propionate metabolism, which is relevant to propionic acidemia. In parasites, it is important for infection.
How can I study propionate CoA-transferase activity in the lab?
Common methods include enzyme activity assays, site-directed mutagenesis, heterologous expression, and CRISPR knockout in model organisms.
What CRISPR models are available for propionate CoA-transferase research?
EDITGENE offers knockout, point mutation, knock-in, and overexpression models in various cell types, as well as CRISPR library screening for pathway analysis.
Conclusion
Propionate CoA-transferase activity (GO:0018729) is a well-defined molecular function that plays a central role in propionate metabolism and fermentation. Its mechanism, involving a covalent enzyme-CoA intermediate, is conserved among CoA-transferases. The enzyme is found in diverse organisms, from anaerobic bacteria to parasites, and has biotechnological potential for the production of green chemicals. Understanding its regulation and function can aid in the development of new drugs and metabolic engineering strategies. EDITGENE provides the tools to study this enzyme through precise CRISPR genome editing.
References
- 1. Volodina E et al.. 2014. Characterization of propionate CoA-transferase from Ralstonia eutropha H16.. Appl Microbiol Biotechnol 98(8):3579-89 PMID: 24057402
- 2. Saz HJ et al.. 1994. 2-Methylbutyryl-CoA: succinate acyl-CoA transferase activity and function in Ascaris suum muscle.. Comp Biochem Physiol Biochem Mol Biol 108(4):513-9 PMID: 7953070
- 3. Selmer T et al.. 2002. Propionate CoA-transferase from Clostridium propionicum. Cloning of gene and identification of glutamate 324 at the active site.. Eur J Biochem 269(1):372-80 PMID: 11784332
- 4. Lee JW et al.. 2019. Microbial biosynthesis of lactate esters.. Biotechnol Biofuels 12:226 PMID: 31548868
- 5. van Grinsven KW et al.. 2009. Acetate:succinate CoA-transferase in the anaerobic mitochondria of Fasciola hepatica.. Mol Biochem Parasitol 164(1):74-9 PMID: 19103231
- 6. Bielefeld DR et al.. 1985. Inhibition of carnitine palmitoyl-CoA transferase activity and fatty acid oxidation by lactate and oxfenicine in cardiac muscle.. J Mol Cell Cardiol 17(6):619-25 PMID: 3927008
- 7. McMillin JB et al.. 1992. Evidence for malonyl-CoA-sensitive carnitine acyl-CoA transferase activity in sarcoplasmic reticulum of canine heart.. J Mol Cell Cardiol 24(3):259-68 PMID: 1625348
- 8. McLaughlin GL et al.. 1986. Purification and properties of an acyl CoA transferase from Ascaris suum muscle mitochondria.. Comp Biochem Physiol B 83(3):523-7 PMID: 3456879
