GO:0016701 oxidoreductase activity, acting on single donors with incorporation of molecular oxygen: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0016701 describes a molecular function in which a single electron donor is oxidized and molecular oxygen is incorporated into the substrate, as defined by QuickGO.
• This activity is carried out by oxygenases, including monooxygenases and dioxygenases, that use cofactors such as heme, flavin, or metal ions to activate molecular oxygen.
• Genes annotated with GO:0016701 are involved in diverse pathways, from amino acid metabolism to antibiotic biosynthesis and xenobiotic degradation.
• Dysregulation of oxygenase activity has been linked to cancer, metabolic disorders, and nematode resistance in livestock.
• CRISPR-based models, including knockout, point mutation, and knock-in, are essential to dissect the catalytic and regulatory roles of these enzymes.
• EDITGENE provides end-to-end services for functional validation of GO:0016701-related genes, from library screening to bioinformatics.
Description
GO:0016701, oxidoreductase activity, acting on single donors with incorporation of molecular oxygen, is a molecular function term that captures a fundamental class of redox reactions in which molecular oxygen is directly incorporated into a substrate. This activity is central to many biological processes, including the biosynthesis of signaling molecules, detoxification of xenobiotics, and the metabolism of amino acids and lipids. Researchers studying this term are often interested in how oxygenases contribute to cellular homeostasis and how their dysfunction leads to disease. The QuickGO definition states that catalysis occurs via transfer of hydrogen or electrons from a single donor, with molecular oxygen incorporated into the donor. This distinguishes it from other oxidoreductases that do not incorporate oxygen into the product. Understanding the mechanistic and regulatory features of GO:0016701 is therefore critical for both basic biology and therapeutic development.
oxidoreductase activity, acting on single donors with incorporation of molecular oxygen At A Glance
| GO ID | GO:0016701 |
|---|---|
| GO term | oxidoreductase activity, acting on single donors with incorporation of molecular oxygen |
| Ontology | molecular_function |
| Synonym | oxygenase; oxidoreductase activity, acting on single donors with incorporation of molecular oxygen, miscellaneous |
| Major function | Catalysis of redox reactions where molecular oxygen is incorporated into a donor substrate |
| Cofactors | Often heme, flavin, or metal ions (e.g., Fe, Cu) |
| Substrates | Amino acids, lipids, xenobiotics, and secondary metabolites |
| Related terms | Monooxygenase activity (GO:0004497), dioxygenase activity (GO:0051213) |
What Is GO:0016701?
According to QuickGO, GO:0016701 is defined as catalysis of an oxidation-reduction (redox) reaction in which hydrogen or electrons are transferred from one donor, and molecular oxygen is incorporated into a donor. In simpler terms, it is an oxygenase activity where one substrate is oxidized and oxygen becomes part of the product. The term is synonymous with 'oxygenase' and 'oxidoreductase activity, acting on single donors with incorporation of molecular oxygen, miscellaneous'. This activity requires molecular oxygen as a co-substrate and often involves metal or organic cofactors to activate O2.
Why Is oxidoreductase activity, acting on single donors with incorporation of molecular oxygen Important in Cell Biology?
GO:0016701 is important because oxygen-incorporating redox reactions are essential for numerous physiological and pathological processes. These enzymes participate in the metabolism of drugs and toxins, the synthesis of hormones and neurotransmitters, and the response to oxidative stress. In livestock, genetic variants in oxygenase-related genes have been associated with gastrointestinal nematode resistance, highlighting their role in immune defense. In human health, dysregulated oxygenase activity contributes to cancer progression and metabolic diseases, making these enzymes attractive drug targets.
• Enables the incorporation of molecular oxygen into organic substrates, a key step in biosynthesis and biodegradation.
• Plays a central role in amino acid catabolism, including the degradation of tryptophan and phenylalanine.
• Involved in the biosynthesis of antibiotics and other secondary metabolites in microorganisms.
• Contributes to xenobiotic detoxification and drug metabolism in the liver and other tissues.
• Associated with ferroptosis-related gene signatures in breast cancer, influencing patient survival.
• Linked to gastrointestinal nematode resistance in sheep, with implications for livestock breeding.
• Requires precise cofactor assembly, as shown for quinone cofactor biogenesis in methylamine dehydrogenase.
• Serves as a target for mechanistic studies using molecular modeling and site-directed mutagenesis.
• Provides a basis for CRISPR-based functional genomics to identify causal variants.
• Offers opportunities for enzyme engineering in biocatalysis and industrial applications.
What Happens During oxidoreductase activity, acting on single donors with incorporation of molecular oxygen?
Substrate Binding and Oxygen Activation
In simple terms: The enzyme grabs its substrate and activates oxygen so it can be inserted.
The reaction begins with binding of the donor substrate to the enzyme active site. Molecular oxygen then binds and is activated by a cofactor, such as heme or a metal ion, to form a reactive oxygen species. This activation is critical for subsequent oxygen incorporation.
Oxygen Incorporation and Product Formation
In simple terms: Oxygen is added to the substrate, changing it into a new product.
Once activated, one atom of molecular oxygen is incorporated into the substrate, while the other may be reduced to water. This step often involves a transient intermediate, such as a peroxo or oxo species, and results in the oxidized product.
Cofactor Regeneration and Catalytic Cycle
In simple terms: The enzyme resets itself to start another round.
After product release, the cofactor must be regenerated to its active form. This may involve electron transfer from a reductase domain or another redox partner, as seen in non-ribosomal peptide synthetases. The catalytic cycle then repeats.
Regulation by Substrate Availability and Redox State
In simple terms: The speed of the reaction depends on how much substrate is present and the cell's redox balance.
Enzyme activity is modulated by substrate concentration, oxygen availability, and the cellular redox environment. Post-translational modifications and protein-protein interactions can also influence catalytic efficiency.
Key Genes Involved in GO:0016701 oxidoreductase activity, acting on single donors with incorporation of molecular oxygen
The following genes encode enzymes with GO:0016701 activity or are directly involved in its regulation, as supported by published literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| MauG | Quinone cofactor biogenesis in methylamine dehydrogenase | Model for heme-dependent oxygen activation |
| Putrescine oxidase | Oxidation of putrescine with oxygen incorporation | Biocatalyst engineering for industrial applications |
| Tryptophan 2,3-dioxygenase | Catabolism of tryptophan | Drug target in cancer and neurological disorders |
| Indoleamine 2,3-dioxygenase | Immune regulation via tryptophan metabolism | Cancer immunotherapy target |
| Phenylalanine hydroxylase | Conversion of phenylalanine to tyrosine | Model for metabolic disorders |
| Tyrosine hydroxylase | Catecholamine biosynthesis | Neurodegeneration research |
| Lipoxygenase | Lipid peroxidation | Inflammation and cancer studies |
| Cyclooxygenase | Prostaglandin synthesis | Inflammation and pain research |
| Ferroptosis-related genes | Lipid peroxidation and cell death | Breast cancer survival prediction |
| Nematode resistance genes | Immune response in sheep | Livestock breeding |
| Conception rate loci | Reproductive traits in beef heifers | Genetic selection |
| Non-ribosomal peptide synthetase reductase domain | Peptidyl thioester reduction | Antibiotic biosynthesis |
| Methylamine dehydrogenase | Oxidation of methylamine | Bacterial metabolism |
| Quinone cofactor | Electron transfer | Enzyme mechanism |
| Heme oxygenase | Heme degradation | Oxidative stress response |
| Nitric oxide synthase | Nitric oxide synthesis | Vascular and immune function |
| Alkane monooxygenase | Hydrocarbon degradation | Bioremediation |
How Is oxidoreductase activity, acting on single donors with incorporation of molecular oxygen Regulated?
The activity of enzymes with GO:0016701 is regulated at multiple levels. Transcriptional control often responds to substrate availability or stress signals. Post-translational modifications, such as phosphorylation, can modulate catalytic activity. Cofactor availability, including heme and metal ions, is a critical determinant of enzyme function. In addition, redox-sensitive transcription factors like Nrf2 can induce antioxidant and detoxifying oxygenases.
oxidoreductase activity, acting on single donors with incorporation of molecular oxygen and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| Indoleamine 2,3-dioxygenase | Cancer immune evasion | Knockout in cancer cell lines |
| Phenylalanine hydroxylase | Phenylketonuria | Point mutation knock-in in mice |
| Lipoxygenase | Inflammation and cancer | Overexpression in epithelial cells |
| MauG | Bacterial metabolism | Knockout in Methylobacterium |
| Nematode resistance genes | Parasite resistance in sheep | GWAS and functional validation |
Cancer
Oxygenases such as indoleamine 2,3-dioxygenase and lipoxygenases are frequently dysregulated in cancer. Their activity can promote immune evasion and tumor progression, making them targets for therapy. Ferroptosis-related genes, many of which are oxygenases, have been used to predict overall survival in breast cancer.
Metabolic Disorders
Defects in phenylalanine hydroxylase, a classic GO:0016701 enzyme, cause phenylketonuria. Other oxygenases are involved in lipid metabolism and energy homeostasis, linking them to obesity and diabetes.
Neurodegeneration
Tryptophan and tyrosine hydroxylases are critical for neurotransmitter synthesis. Their dysfunction has been implicated in Parkinson's disease and mood disorders.
Infectious and Parasitic Diseases
In livestock, genetic variants in oxygenase-related genes are associated with resistance to gastrointestinal nematodes, suggesting a role in immune defense. In humans, oxygenases contribute to the metabolism of antimicrobial drugs.
From oxidoreductase activity, acting on single donors with incorporation of molecular oxygen-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of oxygenase activity affect tumor growth? | Knockout cell line or mouse model |
| Does a specific point mutation alter catalytic efficiency? | Point mutation knock-in via CRISPR |
| Can a tagged oxygenase be used for localization studies? | Knock-in of fluorescent tag |
| Does overexpression of an oxygenase induce ferroptosis? | Overexpression cell line |
| Which cofactors are essential for enzyme function? | Knockout of cofactor biosynthesis genes |
| Can oxygenase activity be engineered for biocatalysis? | Directed evolution and overexpression |
How to Study the oxidoreductase activity, acting on single donors with incorporation of molecular oxygen Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Oxygen consumption assay | Rate of oxygen incorporation | Enzyme kinetics |
| LC-MS/MS | Product formation | Metabolite identification |
| CRISPR knockout screen | Gene essentiality and resistance | Functional genomics |
| RNA-seq | Transcript abundance | Expression profiling |
| Molecular modeling | Substrate binding and catalysis | Mechanistic studies |
| Site-directed mutagenesis | Effect of point mutations | Structure-function analysis |
| Immunoblotting | Protein expression and modification | Regulation studies |
| Fluorescence microscopy | Subcellular localization | Tagged knock-in |
Enzyme Activity Assays
Direct measurement of oxygen consumption or product formation using spectrophotometric or chromatographic methods is the gold standard for assessing GO:0016701 activity.
CRISPR Screening
Genome-wide CRISPR knockout or activation screens can identify genes that modulate oxygenase activity or sensitivity to oxidative stress.
Structural and Computational Modeling
Molecular modeling and crystallography provide insights into substrate binding and catalytic mechanisms, as demonstrated for non-ribosomal peptide synthetases.
Omics Profiling
Transcriptomics and proteomics can reveal expression patterns of oxygenases across tissues and conditions, linking them to disease phenotypes.
How CRISPR Can Be Used to Study GO:0016701 oxidoreductase activity, acting on single donors with incorporation of molecular oxygen
Knockout
CRISPR knockout of oxygenase genes can abolish enzyme activity, allowing researchers to study loss-of-function phenotypes in cancer, metabolism, and immunity.
Point Mutation
Introducing specific point mutations via CRISPR base editing or HDR can mimic disease-associated variants and dissect catalytic residues.
Knock-in
Knock-in of tags or reporter genes enables real-time tracking of oxygenase expression and localization.
Overexpression
CRISPR activation or cDNA overexpression can elevate oxygenase levels to study gain-of-function effects, such as ferroptosis induction.
How EDITGENE Supports oxidoreductase activity, acting on single donors with incorporation of molecular oxygen Research
Researchers studying oxidoreductase activity, acting on single donors with incorporation of molecular oxygen-related genes often need to determine whether a candidate gene is causally involved in a specific phenotype. EDITGENE provides a comprehensive suite of CRISPR services to accelerate this functional validation.
Contact EDITGENE today to design your custom CRISPR model for oxidoreductase activity, acting on single donors with incorporation of molecular oxygen research.
Frequently Asked Questions About oxidoreductase activity, acting on single donors with incorporation of molecular oxygen
What is GO:0016701?
GO:0016701 is a Gene Ontology molecular function term for oxidoreductase activity, acting on single donors with incorporation of molecular oxygen, also known as oxygenase activity.
What genes are involved in oxidoreductase activity, acting on single donors with incorporation of molecular oxygen?
Genes include MauG, putrescine oxidase, tryptophan 2,3-dioxygenase, indoleamine 2,3-dioxygenase, and many others.
What diseases are associated with GO:0016701?
Dysregulation of oxygenases is linked to cancer, metabolic disorders, neurodegeneration, and infectious diseases.
How can I study oxidoreductase activity, acting on single donors with incorporation of molecular oxygen?
Common methods include enzyme activity assays, CRISPR knockout screens, and omics profiling.
What are the cofactors for GO:0016701 enzymes?
Many require heme, flavin, or metal ions such as iron or copper.
Is GO:0016701 the same as monooxygenase activity?
No, GO:0016701 is broader and includes both monooxygenases and dioxygenases that incorporate molecular oxygen.
Can CRISPR be used to study GO:0016701?
Yes, CRISPR knockout, point mutation, and knock-in models are widely used to dissect oxygenase function.
What is the role of oxygenases in cancer?
They can promote immune evasion and tumor progression, and ferroptosis-related oxygenases predict breast cancer survival.
How does EDITGENE support GO:0016701 research?
EDITGENE offers knockout, point mutation, knock-in, overexpression, library screening, and bioinformatics services.
What model organisms are used to study GO:0016701?
Common models include human cell lines, mice, sheep, and bacteria such as Methylobacterium.
Conclusion
GO:0016701 represents a vital class of oxygen-incorporating redox enzymes that impact metabolism, immunity, and disease. Understanding their mechanisms and regulation is essential for developing targeted therapies and biotechnological applications. EDITGENE's CRISPR services provide robust tools to functionally validate these enzymes in any biological context.
References
- 1. Oliver KF et al.. 2020. Loci associated with conception rate in crossbred beef heifers.. PLoS One 15(4):e0230422 PMID: 32271764
- 2. Jin LY et al.. 2021. The role of ferroptosis-related genes for overall survival prediction in breast cancer.. J Clin Lab Anal 35(12):e24094 PMID: 34741349
- 3. Kamathewatta NJB et al.. 2020. Self-Immobilized Putrescine Oxidase Biocatalyst System Engineered with a Metal Binding Peptide.. Langmuir 36(40):11908-11917 PMID: 32921059
- 4. Stafuzza NB et al.. 2023. Weighted single-step genome-wide association study and functional enrichment analyses for gastrointestinal nematode resistance traits in Santa Ines sheep.. Vet Parasitol 323:110047 PMID: 37857178
- 5. Manavalan B et al.. 2010. Molecular modeling of the reductase domain to elucidate the reaction mechanism of reduction of peptidyl thioester into its corresponding alcohol in non-ribosomal peptide synthetases.. BMC Struct Biol 10:1 PMID: 20067617
- 6. Pearson AR et al.. 2004. Further insights into quinone cofactor biogenesis: probing the role of mauG in methylamine dehydrogenase tryptophan tryptophylquinone formation.. Biochemistry 43(18):5494-502 PMID: 15122915
