GO:0016709 oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H 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:0016709 describes a molecular function in which NADH or NADPH supplies electrons to a paired donor while one atom of molecular oxygen is incorporated into the substrate.
• This activity is a subset of monooxygenase chemistry and is central to oxidative metabolism, xenobiotic detoxification, and lipid and sterol modification.
• Genes annotated with this function include cytochrome P450 family members, flavin-dependent monooxygenases, and other NAD(P)H-dependent oxygenases.
• Dysregulation of GO:0016709 enzymes is linked to cancer progression, metabolic disorders, and altered drug responses.
• CRISPR knockout, point-mutation, knock-in, and overexpression models are essential to assign causality to individual GO:0016709 genes.
• Functional enrichment of GO:0016709 appears in genome-wide association and transcriptomic studies across species, from livestock to human tumors.
Description
GO:0016709 is a Gene Ontology molecular function term that defines a specific class of oxidation-reduction reactions. In these reactions, NADH or NADPH acts as one electron donor, a second donor is oxidized in a paired fashion, and one atom of molecular oxygen is incorporated into the product. This distinguishes GO:0016709 from simple oxidases or dehydrogenases, which do not incorporate oxygen into the substrate. The term is therefore a precise annotation for monooxygenase-type chemistry that depends on reduced pyridine nucleotides. Researchers encounter GO:0016709 in functional enrichment analyses of transcriptomic and genomic datasets. For example, weighted single-step genome-wide association studies in sheep identified enrichment of oxidoreductase activities, including NAD(P)H-dependent oxygen incorporation, in traits related to gastrointestinal nematode resistance. Integrated epigenomic profiling in renal cell carcinoma revealed endogenous retrovirus reactivation associated with altered expression of genes carrying this activity. Loci associated with conception rate in crossbred beef heifers also mapped to genes with oxidoreductase functions. These examples show that GO:0016709 is not an abstract annotation but a recurring functional theme in physiology and disease. Because GO:0016709 enzymes participate in hormone synthesis, fatty acid desaturation, and detoxification, they are high-value targets for both basic research and therapeutic development. Understanding which genes carry this activity, how they are regulated, and how they contribute to disease requires precise experimental models. This article summarizes the definition, mechanism, key genes, disease links, and research methods for GO:0016709, with a focus on CRISPR-based approaches for causal validation.
oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen At A Glance
| GO ID | GO:0016709 |
|---|---|
| GO term | oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen |
| Ontology | molecular_function |
| Synonym | oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NADH or NADPH as one donor, and incorporation of one atom of oxygen |
| Major function | NAD(P)H-dependent monooxygenation with incorporation of one oxygen atom into the substrate |
| Electron donor | NADH or NADPH |
| Oxygen fate | One atom of molecular oxygen is incorporated into the product; the other is typically reduced to water |
| Representative enzymes | Cytochrome P450 monooxygenases, flavin-dependent monooxygenases, and related NAD(P)H-dependent oxygenases |
| Related GO terms | Monooxygenase activity (GO:0004497); oxidoreductase activity (GO:0016491) |
What Is GO:0016709?
In simple terms, GO:0016709 describes enzymes that use NADH or NADPH to donate electrons while inserting one oxygen atom from molecular oxygen into another molecule. The QuickGO definition states: Catalysis of an oxidation-reduction (redox) reaction in which hydrogen or electrons are transferred from NADH or NADPH and one other donor, and one atom of oxygen is incorporated into one donor. This means the reaction consumes NAD(P)H, consumes molecular oxygen, and produces a hydroxylated or oxygenated product plus water. The term is a molecular function, not a biological process or cellular component, and it is often used to annotate cytochrome P450 monooxygenases, flavin-containing monooxygenases, and related enzymes.
Why Is oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen Important in Cell Biology?
GO:0016709 is important because it defines a catalytic mechanism that is essential for biosynthesis, detoxification, and metabolic homeostasis. Enzymes with this activity modify lipids, steroids, vitamins, and xenobiotics, and they are frequently dysregulated in cancer and metabolic disease. In livestock and agricultural genomics, enrichment of this activity has been associated with parasite resistance and reproductive traits, indicating its broad biological relevance. Because the reaction consumes NAD(P)H and molecular oxygen, it directly links cellular redox state to product formation, making it a sensitive node for metabolic regulation. For researchers, GO:0016709 provides a precise functional label for interpreting enrichment results and for prioritizing genes for CRISPR validation.
• GO:0016709 enzymes catalyze NAD(P)H-dependent oxygen incorporation, a core reaction in oxidative metabolism.
• They are involved in the metabolism of steroids, fatty acids, and xenobiotics, affecting drug efficacy and toxicity.
• Enrichment of this activity has been reported in cancer epigenomic and transcriptomic studies, including renal cell carcinoma.
• Genome-wide association studies in livestock have linked oxidoreductase activities to nematode resistance and conception rate.
• The activity is dependent on cellular redox balance because it consumes NAD(P)H.
• It is a common annotation in plant and microbial genomes, where it contributes to secondary metabolism and stress responses.
• Dysregulation can alter hormone and lipid signaling, contributing to metabolic and proliferative diseases.
• CRISPR knockout and knock-in models allow direct testing of whether a specific GO:0016709 gene drives a phenotype.
• Functional enrichment of GO:0016709 can serve as a biomarker signature in transcriptomic studies.
• Understanding this activity supports the development of enzyme inhibitors and prodrug activation strategies.
Molecular Mechanism of oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen
NAD(P)H Binding and Electron Transfer
In simple terms: The enzyme first grabs NADH or NADPH to get electrons.
The reaction begins with binding of NADH or NADPH to the enzyme active site. The reduced pyridine nucleotide donates two electrons, which are transferred through a flavin or heme cofactor to the catalytic center. This step is essential because it primes the enzyme for oxygen activation. In many annotated enzymes, NAD(P)H binding induces conformational changes that position the second donor for catalysis.
Oxygen Activation and Incorporation
In simple terms: One oxygen atom from O2 is inserted into the substrate.
After electron transfer, molecular oxygen binds to the reduced cofactor and is activated. One atom of oxygen is incorporated into the substrate, while the other is typically reduced to water. This monooxygenase chemistry distinguishes GO:0016709 from oxidases that do not incorporate oxygen. The reaction is dependent on the availability of molecular oxygen and reducing equivalents.
Paired Donor Oxidation
In simple terms: A second molecule is oxidized at the same time.
The term specifies paired donors, meaning that NAD(P)H and another donor are both oxidized. The second donor can be a lipid, steroid, xenobiotic, or other small molecule. The coupling of electron transfer from NAD(P)H to substrate oxygenation is often imperfect, leading to uncoupling and reactive oxygen species production under certain conditions.
Product Release and Enzyme Turnover
In simple terms: The oxygenated product is released and the enzyme resets.
Following oxygen incorporation, the hydroxylated or oxygenated product is released, and the enzyme returns to its resting state. Turnover requires reoxidation of the cofactor and, in some cases, interaction with redox partners such as cytochrome P450 reductase. The catalytic cycle is sensitive to substrate availability and redox balance, which influences overall flux through GO:0016709-dependent pathways.
Key Genes Involved in GO:0016709 oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen
The following genes and gene families are representative carriers of GO:0016709 activity, based on functional annotations and published studies.
| Gene | Major Role | Research Relevance |
|---|---|---|
| CYP1A1 | Cytochrome P450 monooxygenase; xenobiotic metabolism | Studied in cancer and detoxification |
| CYP1B1 | Cytochrome P450 monooxygenase; estrogen metabolism | Linked to hormone-dependent cancers |
| CYP2E1 | Cytochrome P450 monooxygenase; alcohol and drug metabolism | Model for oxidative stress and liver injury |
| CYP3A4 | Cytochrome P450 monooxygenase; drug metabolism | Key enzyme in pharmacokinetics |
| CYP17A1 | Steroidogenic monooxygenase | Target in prostate cancer and hormone disorders |
| CYP19A1 | Aromatase; estrogen biosynthesis | Relevant to breast cancer and reproduction |
| FMO1 | Flavin-containing monooxygenase | Xenobiotic and drug oxidation |
| FMO3 | Flavin-containing monooxygenase | Metabolism of trimethylamine and drugs |
| SQLE | Squalene monooxygenase; cholesterol synthesis | Metabolic and cancer studies |
| SCD | Stearoyl-CoA desaturase; fatty acid desaturation | Lipid metabolism and metabolic disease |
| ALOX5 | Arachidonate 5-lipoxygenase | Inflammation and cancer |
| ALOX15 | Arachidonate 15-lipoxygenase | Ferroptosis and cancer |
| PTGS1 | Prostaglandin-endoperoxide synthase 1 | Inflammation and drug targeting |
| PTGS2 | Prostaglandin-endoperoxide synthase 2 | Inflammation and cancer |
| NOS2 | Inducible nitric oxide synthase | Immune response and oxidative stress |
| NOS3 | Endothelial nitric oxide synthase | Vascular function |
| KMO | Kynurenine 3-monooxygenase | Neurodegeneration and inflammation |
| TDO2 | Tryptophan 2,3-dioxygenase | Cancer metabolism and immune evasion |
How Is oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen Regulated?
GO:0016709 activity is regulated at multiple levels. Transcriptional control of cytochrome P450 and flavin-containing monooxygenase genes responds to xenobiotic sensors such as AhR and PXR. Post-translational regulation includes phosphorylation and interaction with redox partners like cytochrome P450 reductase, which supplies electrons. Cellular redox state, NAD(P)H availability, and oxygen tension directly influence catalytic flux. In cancer, epigenetic changes and endogenous retrovirus reactivation can alter expression of oxidoreductase genes, as shown in renal cell carcinoma. Hormonal and metabolic signals also modulate steroidogenic enzymes carrying this activity.
oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| CYP1B1 | Hormone-dependent cancers | Knockout in cancer cell lines |
| ALOX15 | Ferroptosis and breast cancer | Point mutation to alter catalytic activity |
| SQLE | Cholesterol metabolism and metabolic disease | Overexpression in hepatocytes |
| CYP17A1 | Prostate cancer and steroid disorders | Knock-in of patient variants |
| FMO3 | Trimethylaminuria and drug metabolism | Knockout in liver cells |
Cancer and Oxidoreductase Dysregulation
GO:0016709 enzymes are frequently altered in cancer. Integrated epigenomic profiling in renal cell carcinoma revealed endogenous retrovirus reactivation and altered expression of oxidoreductase genes. Ferroptosis-related genes, including lipoxygenases with this activity, have been used to predict overall survival in breast cancer. Cytochrome P450 enzymes influence drug metabolism and can affect chemotherapy outcomes. These findings suggest that GO:0016709 activity contributes to tumor biology through oxidative stress, lipid peroxidation, and hormone synthesis.
Metabolic and Reproductive Disorders
Genes carrying GO:0016709 activity participate in steroidogenesis and lipid metabolism. Loci associated with conception rate in crossbred beef heifers include oxidoreductase genes, indicating a role in reproductive physiology. Squalene monooxygenase and stearoyl-CoA desaturase are involved in cholesterol and fatty acid synthesis, linking this activity to metabolic disorders. Dysregulation can alter hormone levels and membrane lipid composition, contributing to disease risk.
Infectious and Parasitic Disease
In livestock, enrichment of oxidoreductase activities has been associated with gastrointestinal nematode resistance in Santa Ines sheep. This suggests that GO:0016709 enzymes may influence immune or metabolic responses to parasites. In bees, baicalin treatment affected Nosema ceranae proliferation, with potential involvement of oxidative metabolism. These examples highlight the broad relevance of this activity beyond human disease.
From oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Is a candidate GO:0016709 gene required for substrate oxygenation? | CRISPR knockout cell line |
| Does a specific amino acid change alter catalytic activity? | Point-mutation knock-in |
| Can a disease-associated variant recapitulate a phenotype? | Knock-in of the variant allele |
| Where is the enzyme localized in the cell? | Tagged knock-in with fluorescent protein |
| Does overexpression drive metabolic flux? | Overexpression cell model |
| Which pathways depend on this activity? | CRISPR library screening |
How to Study the oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen Process
| Method | What It Measures | Typical Application |
|---|---|---|
| RNA-seq | Gene expression changes | Enrichment of GO:0016709 genes |
| CRISPR knockout | Loss-of-function phenotype | Causal gene validation |
| Point mutation | Effect of specific amino acid change | Catalytic mechanism studies |
| Knock-in | Disease variant function | Variant modeling |
| Overexpression | Gain-of-function effects | Metabolic flux analysis |
| Enzymatic assay | NAD(P)H consumption and product formation | Kinetic characterization |
| Metabolomics | Oxygenated product levels | Pathway flux measurement |
| Proteomics | Protein abundance and interactions | Enzyme complex analysis |
Transcriptomic and Enrichment Analysis
RNA sequencing followed by Gene Ontology enrichment is a standard approach to identify GO:0016709 signatures. Studies in sheep and cattle used genome-wide association and functional enrichment to link oxidoreductase activities to traits. In cancer, integrated epigenomic profiling revealed expression changes in oxidoreductase genes. These methods help prioritize candidate genes for functional validation.
CRISPR-Based Functional Genomics
CRISPR knockout, point mutation, knock-in, and overexpression models allow direct testing of GO:0016709 gene function. Pooled CRISPR screens can identify which oxidoreductase genes are required for cell growth or drug response. These approaches are essential for moving from correlation to causation.
Biochemical and Enzymatic Assays
Enzymatic assays measure NAD(P)H consumption and oxygen incorporation. Electrochemical and spectroscopic methods have been used to characterize bilirubin oxidase adlayers, providing a framework for studying oxygen-dependent redox enzymes. Such assays can determine kinetic parameters and cofactor requirements for GO:0016709 enzymes.
Proteomics and Metabolomics
Mass spectrometry-based proteomics and metabolomics can quantify enzyme abundance and oxygenated products. These methods are useful for tracing flux through GO:0016709-dependent pathways and for identifying biomarkers. Combining metabolomics with CRISPR models strengthens causal inference.
How CRISPR Can Be Used to Study GO:0016709 oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen
Knockout
CRISPR knockout of a GO:0016709 gene eliminates its catalytic activity, allowing researchers to test whether it is required for a specific metabolic or disease phenotype. For example, knocking out CYP1B1 or ALOX15 can reveal their roles in hormone metabolism or ferroptosis. Knockout models are the first step in causal validation.
Point Mutation
Point mutations can be introduced to alter catalytic residues or cofactor-binding sites. This is useful for dissecting the mechanism of oxygen incorporation and for testing whether a specific amino acid is essential for GO:0016709 activity. Such models complement structural studies.
Knock-in
Knock-in of disease-associated variants or tagged versions of the enzyme allows researchers to study variant function and localization. For example, knocking in a patient-derived CYP17A1 mutation can model steroid disorders. Tagged knock-in with fluorescent proteins enables imaging of enzyme trafficking.
Overexpression
Overexpression of a GO:0016709 gene can drive metabolic flux and reveal gain-of-function phenotypes. Overexpressing SQLE or SCD can increase cholesterol or fatty acid synthesis, respectively. This approach is useful for identifying downstream pathways and potential therapeutic targets.
How EDITGENE Supports oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H 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, NAD(P)H 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 phenotype. EDITGENE provides CRISPR-based cell models and screening services to answer these questions with precision.
Contact EDITGENE today to design your custom CRISPR model for oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H 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, NAD(P)H as one donor, and incorporation of one atom of oxygen
What is GO:0016709?
GO:0016709 is a Gene Ontology molecular function term for oxidoreductase activity that uses NADH or NADPH as one donor and incorporates one atom of molecular oxygen into another donor.
What genes are involved in oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen?
Representative genes include cytochrome P450 family members such as CYP1A1, CYP1B1, and CYP3A4, flavin-containing monooxygenases such as FMO1 and FMO3, and other oxygenases like ALOX15 and SQLE.
What is the difference between GO:0016709 and monooxygenase activity?
GO:0016709 is a specific child of monooxygenase activity that requires NADH or NADPH as one electron donor and incorporates exactly one oxygen atom into the substrate.
Why is GO:0016709 important in cancer?
Enzymes with this activity can produce oxidative stress, modify hormones, and influence drug metabolism, and their expression is altered in cancers such as renal cell carcinoma and breast cancer.
How can CRISPR help study GO:0016709?
CRISPR knockout, point mutation, knock-in, and overexpression models allow researchers to test whether a specific oxidoreductase gene is required for a phenotype.
What diseases are linked to GO:0016709 enzymes?
They have been linked to cancer, metabolic disorders, reproductive traits, and parasitic infections in livestock.
What cofactors are used by GO:0016709 enzymes?
They use NADH or NADPH as electron donors and often require flavin or heme cofactors for catalysis.
How is GO:0016709 activity measured?
Enzymatic assays measure NAD(P)H consumption and oxygen incorporation, often combined with metabolomics or proteomics.
Is GO:0016709 found in plants?
Yes, genome-wide analyses in rice and Arabidopsis have identified sulfur-encoding and oxidoreductase genes with this activity.
What model systems are best for studying GO:0016709?
Cell lines with CRISPR modifications, animal models, and biochemical assays are commonly used, depending on the research question.
Conclusion
GO:0016709 defines a precise and biologically important molecular function: NAD(P)H-dependent oxygen incorporation into a paired donor. Its enzymes participate in metabolism, detoxification, hormone synthesis, and disease, and they are recurrently identified in genomic and transcriptomic studies. Understanding this activity requires integrating enrichment analysis with causal CRISPR models. EDITGENE provides the tools to build knockout, point-mutation, knock-in, and overexpression cell models for GO:0016709 genes, accelerating functional discovery.
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
