GO:0016716 oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, another compound as one donor, and incorporation of one atom of oxygen: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0016716 describes a molecular function in which a single enzyme uses two donors and incorporates one atom of molecular oxygen into one donor, while the other oxygen atom is typically reduced to water.
• This activity is central to oxidative metabolism, including fatty acid desaturation, sterol biosynthesis, and xenobiotic transformation.
• Enzymes with this activity often require cofactors such as FAD, NAD(P)H, and cytochrome P450 heme or non-heme diiron centers.
• Dysregulation of these enzymes is linked to cancer, metabolic disorders, and altered drug responses.
• CRISPR knockout, point-mutation, and knock-in models are essential to assign causal roles to candidate genes with this activity.
• EDITGENE provides end-to-end CRISPR cell model and library screening services to study GO:0016716-related genes.
Description
GO:0016716 is a Gene Ontology molecular function term that defines a specific class of oxidoreductase enzymes. These enzymes catalyze redox reactions in which hydrogen or electrons are transferred from each of two donors, and one atom of molecular oxygen is incorporated into one donor. This distinguishes them from monooxygenases that use a single donor or dioxygenases that incorporate both oxygen atoms. The term is broad enough to include cytochrome P450 enzymes, flavin-dependent monooxygenases, and non-heme iron oxygenases that act on paired donors. Understanding this activity is critical because it underlies the biosynthesis and degradation of numerous biomolecules, including fatty acids, steroids, and drugs. Researchers studying this term often focus on identifying the specific gene products, their cofactor requirements, and their roles in disease. The integration of genomic, biochemical, and CRISPR-based approaches has accelerated the functional annotation of these enzymes.
oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, another compound as one donor, and incorporation of one atom of oxygen At A Glance
| GO ID | GO:0016716 |
|---|---|
| GO term | oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, another compound as one donor, and incorporation of one atom of oxygen |
| Ontology | molecular_function |
| Synonym | none |
| Major function | Catalysis of redox reactions using two donors and incorporating one atom of oxygen into one donor |
| Cofactors | Often requires FAD, NAD(P)H, heme, or non-heme iron |
| Example enzymes | Cytochrome P450 monooxygenases, flavin-dependent monooxygenases, and some non-heme iron oxygenases |
| Biological processes | Fatty acid desaturation, sterol biosynthesis, xenobiotic metabolism |
| Disease relevance | Cancer, metabolic disorders, and altered drug metabolism |
What Is GO:0016716?
According to the QuickGO definition, GO:0016716 refers to catalysis of an oxidation-reduction (redox) reaction in which hydrogen or electrons are transferred from each of two donors, and one atom of oxygen is incorporated into one donor. In other words, the enzyme uses two substrates as electron donors; one oxygen atom from molecular oxygen is inserted into one of the donors, while the other oxygen atom is reduced, usually to water. This activity is distinct from other oxygenase activities because it explicitly requires two donors and incorporates only one oxygen atom into the product.
Why Is oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, another compound as one donor, and incorporation of one atom of oxygen Important in Cell Biology?
GO:0016716 is important because it captures a fundamental catalytic strategy used by nature to introduce oxygen into organic molecules. This activity is essential for the biosynthesis of unsaturated fatty acids, cholesterol, and steroid hormones, as well as for the detoxification of drugs and environmental chemicals. Defects in these enzymes can lead to metabolic diseases, and their overexpression is often observed in cancers, making them attractive therapeutic targets. Moreover, many drugs are metabolized by enzymes with this activity, influencing drug efficacy and toxicity. Therefore, precise annotation and functional characterization of genes with GO:0016716 are critical for both basic biology and translational research.
• Enables the incorporation of molecular oxygen into diverse substrates, a key step in biosynthesis and biodegradation.
• Plays a central role in fatty acid desaturation and membrane lipid homeostasis.
• Required for sterol and steroid hormone biosynthesis.
• Mediates phase I metabolism of xenobiotics, affecting drug half-life and toxicity.
• Dysregulation is associated with cancer, obesity, and cardiovascular diseases.
• Provides targets for herbicide and antibiotic development.
• Involved in the biosynthesis of secondary metabolites in plants and microbes.
• Serves as a model for studying enzyme evolution and catalytic promiscuity.
• Enables the production of valuable chemicals through biocatalysis.
• Its annotation helps interpret genome-wide association studies and functional genomics data.
What Happens During oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, another compound as one donor, and incorporation of one atom of oxygen?
Substrate Binding and Donor Selection
In simple terms: The enzyme grabs two different molecules that will donate electrons.
The reaction begins with the binding of two donors to the enzyme active site. One donor is typically an organic substrate that will receive the oxygen atom, while the other donor is often NAD(P)H or a similar reducing agent. The enzyme must precisely position both donors to ensure that oxygen activation and transfer occur efficiently. Structural studies of cytochrome P450 enzymes show that substrate binding induces conformational changes that facilitate electron transfer from the redox partner.
Oxygen Activation and Incorporation
In simple terms: One oxygen atom from O2 is inserted into the substrate, and the other becomes water.
Molecular oxygen binds to a metal center (e.g., heme iron or non-heme diiron) and is activated by electrons from the second donor. This activation leads to the cleavage of the O-O bond, with one oxygen atom incorporated into the substrate and the other reduced to water. The precise mechanism varies among enzyme families but generally involves high-valent metal-oxo intermediates that abstract hydrogen from the substrate, followed by oxygen rebound.
Product Release and Enzyme Regeneration
In simple terms: The modified substrate leaves, and the enzyme resets for another round.
After oxygen incorporation, the oxygenated product is released from the active site. The enzyme then returns to its resting state, often through reduction by electron transfer partners such as cytochrome P450 reductase or ferredoxin. This regeneration step is crucial for catalytic turnover and is tightly regulated to prevent uncoupling and reactive oxygen species production.
Cofactor and Electron Transfer Chains
In simple terms: Helper molecules shuttle electrons to keep the reaction going.
Many enzymes with GO:0016716 activity rely on electron transfer chains involving FAD, FMN, iron-sulfur clusters, or heme groups. For example, cytochrome P450 enzymes receive electrons from NADPH via cytochrome P450 oxidoreductase. Flavin-dependent monooxygenases use FAD as a cofactor to activate oxygen and transfer it to the substrate. The efficiency of these chains determines the overall catalytic rate and coupling efficiency.
Regulation by Substrate Availability and Post-translational Modifications
In simple terms: The cell controls when and how fast these enzymes work.
The activity of these enzymes can be regulated at multiple levels, including transcriptional induction by xenobiotics, allosteric modulation by metabolites, and post-translational modifications such as phosphorylation. For instance, the activity of some cytochrome P450 enzymes is influenced by phosphorylation, which can alter their interaction with redox partners. Substrate availability and product inhibition also play key roles in fine-tuning flux through these pathways.
Key Genes Involved in GO:0016716 oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, another compound as one donor, and incorporation of one atom of oxygen
The following genes encode enzymes or associated proteins that exhibit or support GO:0016716 activity, based on published literature.
| Gene | Major Role | Research Relevance |
|---|---|---|
| CYP1A1 | Cytochrome P450 monooxygenase involved in xenobiotic metabolism | Studied for activation of procarcinogens and drug metabolism |
| CYP2D6 | Cytochrome P450 monooxygenase metabolizing many drugs | Polymorphisms affect drug response; model for personalized medicine |
| CYP3A4 | Major drug-metabolizing enzyme in liver and intestine | Key for pharmacokinetics and drug-drug interactions |
| FADS1 | Fatty acid desaturase introducing double bonds | Associated with lipid metabolism and cardiovascular risk |
| FADS2 | Fatty acid desaturase involved in PUFA synthesis | Linked to inflammation and metabolic disorders |
| SCD | Stearoyl-CoA desaturase, a delta-9 desaturase | Target for obesity and diabetes research |
| SQLE | Squalene monooxygenase in sterol biosynthesis | Potential target for cholesterol-lowering drugs |
| CYP51A1 | Lanosterol 14-alpha demethylase in sterol biosynthesis | Target of azole antifungals; studied in cancer |
| PTGS1 | Prostaglandin-endoperoxide synthase 1 (COX-1) | Involved in inflammation and pain; drug target |
| PTGS2 | Prostaglandin-endoperoxide synthase 2 (COX-2) | Induced in inflammation and cancer |
| ALOX5 | Arachidonate 5-lipoxygenase | Role in asthma and inflammatory diseases |
| ALOX12 | Arachidonate 12-lipoxygenase | Implicated in cancer and platelet function |
| P450 oxidoreductase (POR) | Electron transfer partner for microsomal P450s | Mutations cause disorders of steroidogenesis |
| FDX1 | Ferredoxin 1, electron donor for mitochondrial P450s | Essential for steroid hormone biosynthesis |
| FDXR | Ferredoxin reductase, transfers electrons from NADPH | Mutations linked to auditory neuropathy |
| CYP11A1 | Cholesterol side-chain cleavage enzyme | Key for steroid hormone synthesis |
| CYP17A1 | 17-alpha-hydroxylase/17,20-lyase | Involved in androgen synthesis and hypertension |
How Is oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, another compound as one donor, and incorporation of one atom of oxygen Regulated?
The activity of enzymes with GO:0016716 is regulated at multiple levels. Transcriptional regulation often involves nuclear receptors such as AhR, CAR, and PXR, which induce cytochrome P450 genes in response to xenobiotics. Post-translational modifications, including phosphorylation and ubiquitination, can modulate enzyme stability and interaction with redox partners. Additionally, substrate availability and product feedback inhibition control flux through metabolic pathways. In some cases, the availability of cofactors such as NADPH and FAD determines catalytic efficiency.
oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, another compound as one donor, and incorporation of one atom of oxygen and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| CYP1A1 | Cancer (procarcinogen activation) | Knockout in HepG2 cells; overexpression in A549 |
| PTGS2 | Inflammation and cancer | Knockout in HCT116; point mutation of catalytic residues |
| FADS1 | Metabolic syndrome | Knockout in HepG2; knock-in of human variant |
| SQLE | Hypercholesterolemia | Knockout in HeLa; overexpression in liver cells |
| CYP2D6 | Drug metabolism variability | Knock-in of polymorphic variants in HepG2 |
Cancer
Altered expression of enzymes with GO:0016716 activity is frequently observed in cancer. For example, overexpression of CYP1A1 and CYP1B1 can activate procarcinogens and promote tumorigenesis. COX-2 (PTGS2) is induced in many cancers and contributes to inflammation and angiogenesis. Targeting these enzymes with inhibitors is an active area of cancer research.
Metabolic Disorders
Dysregulation of fatty acid desaturases (FADS1, FADS2, SCD) is associated with obesity, insulin resistance, and cardiovascular disease. Squalene monooxygenase (SQLE) is a rate-limiting enzyme in cholesterol synthesis and is a potential target for hypercholesterolemia. Mutations in CYP11A1 and CYP17A1 cause disorders of steroidogenesis, leading to hypertension and sexual development abnormalities.
Inflammatory Diseases
Lipoxygenases (ALOX5, ALOX12) and cyclooxygenases (PTGS1, PTGS2) are key players in inflammatory pathways. Their products, leukotrienes and prostaglandins, mediate pain, fever, and asthma. Inhibitors of these enzymes are used to treat inflammatory conditions.
Drug Metabolism and Toxicity
Polymorphisms in CYP2D6 and CYP3A4 affect drug metabolism, leading to variable drug responses and adverse reactions. Understanding these enzymes is crucial for personalized medicine and drug development.
From oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, another compound as one donor, and incorporation of one atom of oxygen-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of gene X reduce enzyme activity? | CRISPR knockout cell line (e.g., HepG2, HEK293T) |
| Does a specific point mutation affect catalytic efficiency? | Point-mutation knock-in via CRISPR |
| Does tagging the endogenous protein alter localization? | Tagged knock-in (e.g., GFP, HA) |
| Does overexpression mimic a disease state? | Overexpression via lentiviral transduction |
| Which genes are essential for the activity? | Genome-wide CRISPR library screening |
| What are the downstream metabolic changes? | Metabolomics combined with knockout models |
How to Study the oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, another compound as one donor, and incorporation of one atom of oxygen Process
| Method | What It Measures | Typical Application |
|---|---|---|
| LC-MS/MS | Substrate and product levels | Quantifying enzyme activity in cell lysates |
| CRISPR knockout | Loss-of-function phenotype | Testing gene essentiality for activity |
| RNA-seq | Transcriptional changes | Identifying induced genes in response to substrates |
| Proteomics | Protein abundance and modifications | Detecting post-translational regulation |
| Metabolomics | Global metabolite profiles | Linking enzyme activity to metabolic pathways |
| Fluorescence microscopy | Subcellular localization | Visualizing tagged enzymes in live cells |
| CRISPR library screening | Genome-wide fitness | Discovering novel regulators of the activity |
Enzymatic Activity Assays
Direct measurement of GO:0016716 activity often uses substrate conversion assays monitored by HPLC, LC-MS, or spectrophotometry. For example, cytochrome P450 activity can be measured using fluorogenic substrates or by monitoring NADPH oxidation. These assays are essential to confirm that a candidate gene encodes a functional enzyme.
CRISPR-Based Functional Genomics
CRISPR knockout and knock-in models allow researchers to test the causal role of specific genes in GO:0016716 activity. Pooled CRISPR screens can identify genes that affect cell fitness under conditions requiring this activity, such as in the presence of specific substrates or inhibitors. These screens are powerful for discovering novel enzymes and regulatory factors.
Transcriptomics and Proteomics
RNA-seq and proteomics can reveal changes in gene expression and protein abundance associated with GO:0016716 activity. For instance, induction of cytochrome P450 genes by xenobiotics can be detected by RNA-seq. Proteomic profiling can identify post-translational modifications that regulate enzyme activity.
Metabolomics and Lipidomics
Metabolomic and lipidomic analyses measure the products of GO:0016716 reactions, such as oxygenated fatty acids or steroids. These methods provide a functional readout of enzyme activity in cells and tissues. They are particularly useful for validating knockout phenotypes.
How CRISPR Can Be Used to Study GO:0016716 oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, another compound as one donor, and incorporation of one atom of oxygen
Knockout
CRISPR knockout is used to completely abolish the expression of a gene encoding an enzyme with GO:0016716 activity. This allows researchers to determine whether the gene is required for a specific metabolic reaction or cellular phenotype. For example, knockout of CYP1A1 in HepG2 cells reduces the metabolism of certain procarcinogens.
Point Mutation
Point mutations can be introduced to alter specific amino acid residues in the catalytic site, allowing structure-function studies. For instance, mutating the heme-binding cysteine in a cytochrome P450 enzyme abolishes activity. These models are valuable for understanding mechanism and for mimicking human polymorphisms.
Knock-in
Knock-in models can introduce tagged versions of the enzyme (e.g., GFP, HA) to study localization and interactions, or to express human variants in a controlled genetic background. This is particularly useful for studying disease-associated mutations.
Overexpression
Overexpression of a gene with GO:0016716 activity can be achieved by lentiviral transduction or CRISPR activation. This is used to study gain-of-function effects, such as increased drug metabolism or production of specific metabolites. Overexpression models are also used in biocatalysis research.
How EDITGENE Supports oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, another compound as one donor, and incorporation of one atom of oxygen Research
Researchers studying oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, another compound as one donor, and incorporation of one atom of oxygen-related genes often need to determine whether a candidate gene is causally involved in a specific metabolic or disease phenotype. EDITGENE provides the necessary CRISPR tools and services to generate precisely engineered cell models for functional validation.
Contact EDITGENE today to design your custom CRISPR model for oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, another compound as one donor, and incorporation of one atom of oxygen research.
Frequently Asked Questions About oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, another compound as one donor, and incorporation of one atom of oxygen
What is GO:0016716?
GO:0016716 is a Gene Ontology molecular function term describing oxidoreductase activity that uses two donors and incorporates one atom of molecular oxygen into one donor.
What genes are involved in GO:0016716?
Genes encoding cytochrome P450 enzymes (e.g., CYP1A1, CYP3A4), fatty acid desaturases (FADS1, FADS2), and cyclooxygenases (PTGS1, PTGS2) are examples.
What diseases are associated with GO:0016716?
Dysregulation is linked to cancer, metabolic disorders, inflammatory diseases, and altered drug metabolism.
How can I study GO:0016716 activity?
Enzymatic assays, CRISPR knockout models, and metabolomics are commonly used to study this activity.
What cofactors are required for GO:0016716?
Many enzymes require FAD, NAD(P)H, heme, or non-heme iron for catalysis.
What is the difference between GO:0016716 and other oxygenase activities?
GO:0016716 specifically requires two donors and incorporates only one oxygen atom, unlike dioxygenases that incorporate both.
Can CRISPR be used to study GO:0016716?
Yes, CRISPR knockout, point mutation, and knock-in models are powerful tools to dissect gene function related to this activity.
What are the typical substrates of GO:0016716 enzymes?
Substrates include fatty acids, steroids, xenobiotics, and prostaglandins.
How is GO:0016716 activity regulated?
It is regulated transcriptionally by nuclear receptors, post-translationally by phosphorylation, and by substrate availability.
What services does EDITGENE offer for GO:0016716 research?
EDITGENE provides CRISPR knockout, point mutation, knock-in, overexpression cell models, library screening, and bioinformatics services.
Conclusion
GO:0016716 represents a fundamental enzymatic activity that is essential for diverse biological processes, from lipid metabolism to drug detoxification. Understanding the genes and mechanisms underlying this activity is crucial for both basic research and therapeutic development. CRISPR-based models, combined with biochemical and omics approaches, offer powerful ways to dissect the function of these enzymes. EDITGENE stands ready to support your research with tailored CRISPR solutions.
References
- 1. 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
- 2. Siebenthall KT et al.. 2019. Integrated epigenomic profiling reveals endogenous retrovirus reactivation in renal cell carcinoma.. EBioMedicine 41:427-442 PMID: 30827930
- 3. Oliver KF et al.. 2020. Loci associated with conception rate in crossbred beef heifers.. PLoS One 15(4):e0230422 PMID: 32271764
- 4. Han X et al.. 2026. Effect of Baicalin on the Proliferation of Nosema ceranae in Apis cerana.. Insects 17(5) PMID: 42188121
- 5. 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
- 6. Abdullah-Zawawi MR et al.. 2022. Genome-wide analysis of sulfur-encoding biosynthetic genes in rice (Oryza sativa L.) with Arabidopsis as the sulfur-dependent model plant.. Sci Rep 12(1):13829 PMID: 35970910
- 7. McArdle T et al.. 2015. Optimizing the Mass-Specific Activity of Bilirubin Oxidase Adlayers through Combined Electrochemical Quartz Crystal Microbalance and Dual Polarization Interferometry Analyses.. ACS Appl Mater Interfaces 7(45):25270-80 PMID: 26506112
