GO:0046577 long-chain-alcohol oxidase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0046577 long-chain-alcohol oxidase activity catalyzes the oxidation of long-chain alcohols to long-chain aldehydes using molecular oxygen, producing water [2,4].
• The term is a molecular_function in the Gene Ontology, with synonyms including fatty alcohol oxidase activity and long-chain fatty alcohol oxidase activity [2,4].
• Enzymes with this activity are found in diverse organisms, from bacteria such as Acinetobacter and Geobacillus to plants like Arabidopsis thaliana and Lotus japonicus, and yeasts such as Candida [1,2,4,5,6,7].
• In Candida yeast, the long-chain fatty alcohol oxidase is a c-type haemoprotein and plays an important role in long-chain fatty acid metabolism.
• Defects in fatty alcohol oxidation are linked to human diseases such as Sjögren-Larsson syndrome, a neurocutaneous disorder caused by mutations in fatty aldehyde dehydrogenase.
• Researchers study this activity using gene knockout, point mutation, knock-in, and overexpression models, combined with biochemical assays and omics methods [1,4,5,8].
Description
Long-chain-alcohol oxidase activity (GO:0046577) is a molecular function that enables the oxidation of long-chain alcohols to their corresponding aldehydes, using molecular oxygen as an electron acceptor and releasing water [2,4]. This activity is part of the broader fatty alcohol metabolism pathway and is critical for the breakdown of long-chain fatty acids and alkanes in various organisms [2,6]. The reaction is formally described as: 2 long-chain alcohol + O2 = 2 long-chain aldehyde + 2 H2O, and it is catalyzed by enzymes known as fatty alcohol oxidases [2,4]. Researchers study this activity to understand lipid metabolism, microbial alkane degradation, and the pathophysiology of inherited disorders of fatty alcohol oxidation [1,3,6]. The enzyme has been characterized in bacteria, plants, and yeasts, revealing diverse structural and catalytic properties [1,2,4,5,7]. In Candida yeast, the long-chain fatty alcohol oxidase is a c-type haemoprotein and plays an important role in long-chain fatty acid metabolism. In plants, AtFao3 from Arabidopsis thaliana is a membrane-bound long-chain alcohol oxidase, and LjFAO1 from Lotus japonicus has been cloned and characterized [4,5]. These findings highlight the evolutionary conservation and functional significance of this activity across kingdoms.
long-chain-alcohol oxidase activity At A Glance
| GO ID | GO:0046577 |
|---|---|
| GO term | long-chain-alcohol oxidase activity |
| Ontology | molecular_function |
| Synonym | fatty alcohol oxidase activity; fatty alcohol:oxygen oxidoreductase activity; long-chain-alcohol:oxygen oxidoreductase activity; long-chain fatty acid oxidase activity; long-chain fatty alcohol oxidase activity |
| Definition | Catalysis of the reaction: 2 long-chain alcohol + O2 = 2 long-chain aldehyde + 2 H2O. |
| Major function | Oxidation of long-chain alcohols to long-chain aldehydes using molecular oxygen. |
| Cofactor | Some enzymes are c-type haemoproteins; others are metal-independent [2,7]. |
| Subcellular location | Membrane-bound in plants; soluble or membrane-associated in other organisms [4,5]. |
| Representative genes | AtFao3 (Arabidopsis thaliana), LjFAO1 (Lotus japonicus), Candida fatty alcohol oxidase, Acinetobacter enzymes [2,4,5,6]. |
What Is GO:0046577?
Long-chain-alcohol oxidase activity (GO:0046577) is defined as the catalysis of the reaction: 2 long-chain alcohol + O2 = 2 long-chain aldehyde + 2 H2O [2,4]. In other words, it is an oxidoreductase activity that converts long-chain alcohols to long-chain aldehydes, using oxygen as the electron acceptor and producing water as a byproduct [2,4]. This activity is synonymous with fatty alcohol oxidase activity, fatty alcohol:oxygen oxidoreductase activity, long-chain-alcohol:oxygen oxidoreductase activity, long-chain fatty acid oxidase activity, and long-chain fatty alcohol oxidase activity [2,4].
Why Is long-chain-alcohol oxidase activity Important in Cell Biology?
Long-chain-alcohol oxidase activity is important because it participates in the metabolism of fatty alcohols and alkanes, which are key processes in microbial degradation of hydrocarbons, plant lipid metabolism, and human inherited disorders of fatty alcohol oxidation [1,2,3,6]. In Candida yeast, the long-chain fatty alcohol oxidase is a c-type haemoprotein and plays an important role in long-chain fatty acid metabolism. In bacteria such as Acinetobacter and Geobacillus, enzymes with this activity are involved in long-chain alkane degradation [1,6,7]. In humans, defects in fatty alcohol oxidation lead to Sjögren-Larsson syndrome, a neurocutaneous disorder characterized by ichthyosis, spasticity, and intellectual disability. Thus, understanding this activity has implications for biotechnology, environmental microbiology, and medicine.
• Enables the conversion of long-chain alcohols to aldehydes, a key step in fatty acid and alkane metabolism [2,6].
• Involved in microbial degradation of alkanes, relevant for bioremediation and biofuel production [1,6,7].
• Plays a role in plant lipid metabolism, as shown for AtFao3 and LjFAO1 [4,5].
• Defects in fatty alcohol oxidation are linked to Sjögren-Larsson syndrome, a human neurocutaneous disorder.
• Provides a target for metabolic engineering of alkane biosynthesis, as demonstrated by introducing a fatty alcohol dehydrogenase.
• Serves as a model for studying c-type haemoprotein chemistry and metal-independent alcohol oxidation [2,7].
• Contributes to the understanding of peroxisomal and membrane-bound lipid oxidation pathways [4,5].
• Offers potential for developing biocatalysts for industrial oxidation reactions [2,4].
What Happens During long-chain-alcohol oxidase activity?
Substrate binding and oxygen activation
In simple terms: The enzyme grabs a long-chain alcohol and oxygen to start the reaction.
The first step involves binding of a long-chain alcohol and molecular oxygen to the active site of the enzyme [2,4]. In c-type haemoproteins such as the Candida fatty alcohol oxidase, the haem iron is involved in oxygen activation. In metal-independent enzymes, different catalytic residues facilitate oxygen activation.
Catalytic oxidation of alcohol to aldehyde
In simple terms: The enzyme removes electrons from the alcohol, turning it into an aldehyde.
The enzyme catalyzes the oxidation of the long-chain alcohol to a long-chain aldehyde, transferring electrons to oxygen [2,4]. This reaction is formally: 2 long-chain alcohol + O2 = 2 long-chain aldehyde + 2 H2O [2,4]. The aldehyde product can be further oxidized to a fatty acid by aldehyde dehydrogenases.
Water formation and product release
In simple terms: Water is made as a byproduct, and the aldehyde is released.
Oxygen is reduced to water, and the aldehyde product is released from the active site [2,4]. The water molecules are byproducts of the reaction [2,4]. The aldehyde can then enter downstream metabolic pathways, such as fatty acid synthesis or degradation [2,6].
Role in long-chain fatty acid metabolism
In simple terms: This reaction helps cells process long-chain fatty acids.
In Candida yeast, the long-chain fatty alcohol oxidase is important for long-chain fatty acid metabolism, linking alcohol oxidation to fatty acid production. In Acinetobacter, long-chain alcohol and aldehyde dehydrogenase activities are involved in alkane degradation. In plants, AtFao3 and LjFAO1 are thought to participate in cuticular wax and lipid metabolism [4,5].
Key Genes Involved in GO:0046577 long-chain-alcohol oxidase activity
The following genes and proteins are representative examples of long-chain-alcohol oxidase activity or related enzymes from bacteria, plants, and yeasts.
| Gene | Major Role | Research Relevance |
|---|---|---|
| AtFao3 (Arabidopsis thaliana) | Membrane-bound long-chain alcohol oxidase | Functional identification in plant lipid metabolism |
| LjFAO1 (Lotus japonicus) | Long-chain fatty alcohol oxidase | Cloning and characterization in legume |
| Candida fatty alcohol oxidase | c-type haemoprotein long-chain fatty alcohol oxidase | Important in long-chain fatty acid metabolism |
| Acinetobacter calcoaceticus alcohol dehydrogenase | Long-chain alcohol dehydrogenase | Involved in alkane degradation |
| Acinetobacter venetianus RAG-1 ADH | NAD+-dependent group III alcohol dehydrogenase | Long-chain alkane degradation |
| Geobacillus thermodenitrificans NG80-2 ADH | Metal-independent long-chain alkyl alcohol dehydrogenase | Novel alcohol dehydrogenases |
| Fatty alcohol dehydrogenase (introduced) | Alcohol dehydrogenase for alkane biosynthesis | Utilizing alcohol for alkane biosynthesis |
| Sjögren-Larsson syndrome FALDH | Fatty aldehyde dehydrogenase | Defects cause neurocutaneous disorder |
| Aldehyde dehydrogenase (Acinetobacter) | Long-chain aldehyde dehydrogenase | Alkane degradation pathway |
| Plant cuticular wax enzymes | Long-chain alcohol oxidation | Wax biosynthesis [4,5] |
| Yeast peroxisomal enzymes | Fatty alcohol oxidation | Lipid metabolism |
| Bacterial alkane monooxygenase | Alkane hydroxylation | Upstream of alcohol oxidation [1,6] |
| Alcohol oxidase (other organisms) | General alcohol oxidation | Comparative enzymology [2,7] |
| Fatty alcohol oxidase homologs | Long-chain alcohol oxidation | Evolutionary studies [4,5] |
| Membrane-bound oxidases | Lipid metabolism | Membrane biology |
| Haem-containing oxidases | Oxygen activation | Enzyme mechanism |
| Metal-independent dehydrogenases | Alcohol oxidation | Biocatalysis |
How Is long-chain-alcohol oxidase activity Regulated?
The regulation of long-chain-alcohol oxidase activity is not fully understood, but some insights exist. In Candida, the enzyme is a c-type haemoprotein and its expression may be regulated by fatty acid availability. In bacteria, genes involved in alkane degradation, including alcohol dehydrogenases, are often regulated by transcriptional regulators in response to alkanes [1,6]. In plants, AtFao3 is membrane-bound and may be regulated developmentally or in response to environmental cues. However, specific regulatory mechanisms such as mTOR or ISR have not been directly linked to this activity in the provided literature.
long-chain-alcohol oxidase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| FALDH (fatty aldehyde dehydrogenase) | Sjögren-Larsson syndrome | Knockout mouse or patient-derived cells |
| Candida fatty alcohol oxidase | Fungal lipid metabolism | Yeast knockout and overexpression |
| Acinetobacter ADH | Alkane degradation | Bacterial knockout [1,6] |
| AtFao3 | Plant lipid metabolism | Arabidopsis knockout |
| LjFAO1 | Legume lipid metabolism | Lotus japonicus knockout |
Sjögren-Larsson syndrome
Sjögren-Larsson syndrome is a neurocutaneous disorder caused by mutations in the fatty aldehyde dehydrogenase gene, leading to impaired oxidation of fatty aldehydes. Although the syndrome primarily involves aldehyde dehydrogenase deficiency, it highlights the importance of the fatty alcohol oxidation pathway in human health. The accumulation of fatty alcohols and aldehydes can cause ichthyosis, spasticity, and intellectual disability.
Metabolic disorders of lipid metabolism
Disorders of fatty alcohol and fatty acid metabolism can lead to various metabolic diseases. The long-chain-alcohol oxidase activity is part of this pathway, and its dysfunction could contribute to lipid accumulation or toxicity [2,6]. However, direct links to specific diseases beyond Sjögren-Larsson syndrome are not well established in the provided literature.
Microbial infections and alkane degradation
In pathogenic bacteria such as Acinetobacter, long-chain alcohol oxidation is involved in alkane degradation, which can be important for survival in host environments [1,6]. Understanding this activity may inform strategies to combat infections or to harness bioremediation [1,6].
From long-chain-alcohol oxidase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does loss of long-chain-alcohol oxidase affect lipid metabolism? | Knockout of AtFao3 in Arabidopsis |
| What is the catalytic mechanism of the c-type haemoprotein? | Point mutations in Candida fatty alcohol oxidase |
| Can the enzyme be redirected to produce aldehydes? | Knock-in of mutant enzyme in yeast |
| Does overexpression increase alkane degradation? | Overexpression of Acinetobacter ADH in E. coli [1,6] |
| What is the role of LjFAO1 in plant development? | Knockout and overexpression in Lotus japonicus |
| Can fatty alcohol dehydrogenase enhance alkane biosynthesis? | Overexpression in microbial hosts |
How to Study the long-chain-alcohol oxidase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| Spectrophotometric oxidase assay | Oxygen consumption or aldehyde formation | Enzyme kinetics [2,4] |
| RNA-seq | Gene expression levels | Regulation studies [1,4,5] |
| qRT-PCR | Transcript abundance | Validation of expression [4,5] |
| Western blot | Protein expression and size | Protein characterization |
| Haem difference spectroscopy | Haem type and redox state | Cofactor identification |
| CRISPR knockout | Loss of function phenotype | Gene function studies [1,4,5] |
| Overexpression | Gain of function phenotype | Biocatalysis and pathway engineering |
| Site-directed mutagenesis | Catalytic residue function | Mechanistic studies [2,7] |
Enzymatic activity assays
Long-chain-alcohol oxidase activity can be measured using spectrophotometric assays that monitor the reduction of oxygen or the formation of aldehyde products [2,4]. These assays typically use long-chain alcohols as substrates and detect the production of NADH or the consumption of oxygen [2,4].
Gene expression analysis
RNA-seq and qRT-PCR can be used to measure the expression of genes encoding long-chain-alcohol oxidases under different conditions [1,4,5]. This helps identify regulatory mechanisms and tissue-specific expression patterns [4,5].
Protein characterization
Proteomics and Western blotting can confirm the presence and size of the enzyme, while spectroscopic methods can characterize haem cofactors. For c-type haemoproteins, difference spectra can identify the haem type.
Genetic manipulation
CRISPR-Cas9 knockout, point mutation, and knock-in models allow functional studies of the enzyme in vivo [1,4,5,8]. Overexpression in heterologous hosts can produce sufficient protein for biochemical studies [2,8].
How CRISPR Can Be Used to Study GO:0046577 long-chain-alcohol oxidase activity
Knockout
CRISPR-Cas9 knockout of genes encoding long-chain-alcohol oxidases can reveal their physiological roles. For example, knocking out AtFao3 in Arabidopsis can help determine its contribution to lipid metabolism. Similarly, knockout of bacterial alcohol dehydrogenases can assess their role in alkane degradation [1,6].
Point Mutation
Point mutations can be introduced to study catalytic residues or cofactor binding sites. For instance, mutating the haem-binding motif in Candida fatty alcohol oxidase can elucidate its mechanism. In metal-independent enzymes, point mutations can identify key residues for catalysis.
Knock-in
Knock-in of tagged or mutant versions of the enzyme can facilitate purification and localization studies. For example, adding a fluorescent tag to AtFao3 can reveal its subcellular localization. Knock-in of a mutant enzyme can also test specific hypotheses about regulation.
Overexpression
Overexpression of long-chain-alcohol oxidase genes in heterologous hosts such as E. coli or yeast can produce large amounts of enzyme for biochemical and structural studies [2,8]. It can also enhance metabolic pathways, such as alkane biosynthesis, as shown by introducing a fatty alcohol dehydrogenase.
How EDITGENE Supports long-chain-alcohol oxidase activity Research
Researchers studying long-chain-alcohol oxidase activity-related genes often need to determine whether a candidate gene is causally involved in lipid metabolism, alkane degradation, or disease. EDITGENE provides comprehensive CRISPR services to create precise cell and animal models for functional studies.
Contact EDITGENE today to design your custom CRISPR model for long-chain-alcohol oxidase activity research.
Frequently Asked Questions About long-chain-alcohol oxidase activity
What is long-chain-alcohol oxidase activity?
Long-chain-alcohol oxidase activity (GO:0046577) is a molecular function that catalyzes the oxidation of long-chain alcohols to long-chain aldehydes using molecular oxygen, producing water [2,4].
What genes are involved in long-chain-alcohol oxidase activity?
Genes include AtFao3 in Arabidopsis thaliana, LjFAO1 in Lotus japonicus, Candida fatty alcohol oxidase, and bacterial alcohol dehydrogenases from Acinetobacter and Geobacillus [1,2,4,5,6,7].
What is the reaction catalyzed by long-chain-alcohol oxidase?
The reaction is: 2 long-chain alcohol + O2 = 2 long-chain aldehyde + 2 H2O [2,4].
Which diseases are associated with long-chain-alcohol oxidase activity?
Defects in the related fatty aldehyde dehydrogenase cause Sjögren-Larsson syndrome, a neurocutaneous disorder.
Is long-chain-alcohol oxidase a haemoprotein?
Some long-chain fatty alcohol oxidases, such as the Candida enzyme, are c-type haemoproteins.
How can I study long-chain-alcohol oxidase activity?
You can use enzymatic assays, gene expression analysis, protein characterization, and CRISPR knockout, point mutation, knock-in, or overexpression models [1,2,4,5,8].
What is the GO ID for long-chain-alcohol oxidase activity?
The GO ID is GO:0046577 [2,4].
What are synonyms for long-chain-alcohol oxidase activity?
Synonyms include fatty alcohol oxidase activity, fatty alcohol:oxygen oxidoreductase activity, long-chain-alcohol:oxygen oxidoreductase activity, long-chain fatty acid oxidase activity, and long-chain fatty alcohol oxidase activity [2,4].
Which organisms have long-chain-alcohol oxidase activity?
This activity is found in bacteria, plants, and yeasts, including Acinetobacter, Arabidopsis thaliana, Lotus japonicus, and Candida [1,2,4,5,6,7].
How does long-chain-alcohol oxidase contribute to alkane degradation?
In bacteria, it oxidizes long-chain alcohols derived from alkanes to aldehydes, which are further metabolized [1,6].
Conclusion
Long-chain-alcohol oxidase activity (GO:0046577) is a fundamental molecular function involved in fatty alcohol and alkane metabolism across bacteria, plants, and yeasts [1,2,4,5,6,7]. Its role in human disease, particularly Sjögren-Larsson syndrome, underscores its biomedical importance. Researchers can leverage CRISPR-based models to dissect its mechanisms and explore biotechnological applications [1,4,5,8]. EDITGENE provides comprehensive services to support these studies.
References
- 1. Chen S et al.. 2024. An NAD(+)-dependent group Ⅲ alcohol dehydrogenase involved in long-chain alkane degradation in Acinetobacter venetianus RAG-1.. Enzyme Microb Technol 172:110343 PMID: 37890395
- 2. Cheng Q et al.. 2005. Candida yeast long chain fatty alcohol oxidase is a c-type haemoprotein and plays an important role in long chain fatty acid metabolism.. Biochim Biophys Acta 1735(3):192-203 PMID: 16046182
- 3. Rizzo WB. 1993. Sjögren-Larsson syndrome.. Semin Dermatol 12(3):210-8 PMID: 8217559
- 4. Cheng Q et al.. 2004. Functional identification of AtFao3, a membrane bound long chain alcohol oxidase in Arabidopsis thaliana.. FEBS Lett 574(1-3):62-8 PMID: 15358540
- 5. Zhao S et al.. 2008. Cloning and characterization of long-chain fatty alcohol oxidase LjFAO1 in lotus japonicus.. Biotechnol Prog 24(3):773-9 PMID: 18396913
- 6. Fox MG et al.. 1992. Long-chain alcohol and aldehyde dehydrogenase activities in Acinetobacter calcoaceticus strain HO1-N.. J Gen Microbiol 138(9):1963-72 PMID: 1402794
- 7. Liu X et al.. 2009. Two novel metal-independent long-chain alkyl alcohol dehydrogenases from Geobacillus thermodenitrificans NG80-2.. Microbiology (Reading) 155(Pt 6):2078-2085 PMID: 19383697
- 8. Sui YA et al.. 2022. Utilizing Alcohol for Alkane Biosynthesis by Introducing a Fatty Alcohol Dehydrogenase.. Appl Environ Microbiol 88(23):e0126422 PMID: 36416567
