GO:0047196 long-chain-alcohol O-fatty-acyltransferase activity: Mechanism, Genes and Research Methods
Research-grade guide for scientists and biopharma professionals
Key Takeaways
• GO:0047196 (long-chain-alcohol O-fatty-acyltransferase activity, also called wax ester synthase or WS/DGAT) catalyzes the esterification of a long-chain alcohol with an acyl-CoA to produce a wax ester and CoA.
• The enzyme is bifunctional in many organisms, exhibiting both wax ester synthase and diacylglycerol acyltransferase (DGAT) activities, which channels acyl groups into neutral lipid storage.
• Wax ester synthases are found in bacteria, plants, and protists; the jojoba (Simmondsia chinensis) enzyme was the first plant wax synthase purified and cloned.
• Substrate specificity varies widely among homologs: some prefer short-chain alcohols, while others favor long-chain or aromatic alcohols, making them tunable for industrial applications.
• Overexpression of WSD1 in Arabidopsis and Camelina increases cuticular wax deposition and improves osmotic stress tolerance, linking the enzyme to plant stress resilience.
• The enzyme is a promising target for metabolic engineering of biofuels, wax esters, and other oleochemicals in microbial and plant hosts.
Description
Long-chain-alcohol O-fatty-acyltransferase activity (GO:0047196) is a molecular function that catalyzes the formation of wax esters from a long-chain alcohol and an acyl-CoA. This reaction is central to the biosynthesis of cuticular waxes in plants, storage lipids in some bacteria, and wax esters in the protist Euglena gracilis. The enzyme is often referred to as wax ester synthase (WS) or wax synthase (WS/DGAT) because many homologs also possess diacylglycerol acyltransferase activity, enabling them to esterify diacylglycerol as well. The first wax synthase was purified and cloned from jojoba (Simmondsia chinensis), where it produces high levels of liquid wax in seeds. Since then, numerous bacterial and plant homologs have been characterized, revealing diverse substrate specificities and physiological roles. Researchers study GO:0047196 to understand lipid metabolism, to engineer organisms for biofuel and oleochemical production, and to improve plant stress tolerance through cuticle modification.
long-chain-alcohol O-fatty-acyltransferase activity At A Glance
| GO ID | GO:0047196 |
|---|---|
| GO term | long-chain-alcohol O-fatty-acyltransferase activity |
| Ontology | molecular_function |
| Synonym | wax ester synthase activity; wax-ester synthase activity; wax synthase activity; acyl-CoA:long-chain-alcohol O-acyltransferase activity |
| Major function | Catalyzes the esterification of long-chain alcohols with acyl-CoA to form wax esters and CoA |
| Reaction | a long-chain-alcohol + acyl-CoA = a long-chain ester + CoA |
| EC number | 2.3.1.75 (not explicitly in QuickGO but commonly associated) |
| Found in | Bacteria, plants, protists (e.g., Marinobacter, Arabidopsis, Euglena, jojoba) |
| Related activity | Diacylglycerol O-acyltransferase activity (DGAT) in bifunctional WS/DGAT enzymes |
What Is GO:0047196?
GO:0047196 describes the catalytic activity of an enzyme that transfers an acyl group from acyl-CoA to a long-chain alcohol, forming a long-chain ester (wax ester) and releasing coenzyme A. The reaction is: long-chain-alcohol + acyl-CoA = long-chain ester + CoA. This activity is synonymous with wax ester synthase, wax-ester synthase, and acyl-CoA:long-chain-alcohol O-acyltransferase. It belongs to the molecular_function ontology and is often associated with the bifunctional wax ester synthase/diacylglycerol acyltransferase (WS/DGAT) family.
Why Is long-chain-alcohol O-fatty-acyltransferase activity Important in Cell Biology?
GO:0047196 is critical for lipid storage and surface barrier formation in many organisms. In plants, wax esters are major components of the cuticle, which protects against desiccation and pathogens. In bacteria and protists, wax esters serve as energy storage compounds and can be engineered for biofuel production. The enzyme's ability to accept diverse substrates makes it a versatile tool for synthetic biology, and its bifunctionality links it to triacylglycerol biosynthesis. Understanding its regulation and specificity is essential for metabolic engineering and for deciphering lipid-related diseases.
• Produces wax esters, which are high-energy storage lipids and industrial feedstocks.
• Contributes to cuticular wax biosynthesis, protecting plants from drought and osmotic stress.
• Bifunctional WS/DGAT enzymes influence triacylglycerol accumulation, relevant to obesity and lipid disorders.
• Enables microbial production of short-chain alkanes and other biofuels when coupled with other enzymes.
• Substrate specificity can be altered for tailored wax ester production.
• Provides a model for studying acyltransferase mechanisms and membrane-bound enzyme catalysis.
• Potential target for engineering crops with improved stress tolerance.
• Used in metagenomic mining for novel biocatalysts.
• Involved in lipid droplet formation in Euglena and other organisms.
• Relevant to industrial production of cosmetics, lubricants, and pharmaceuticals.
What Happens During long-chain-alcohol O-fatty-acyltransferase activity?
Substrate Binding and Acyl Transfer
In simple terms: The enzyme grabs an acyl-CoA and a long-chain alcohol, then joins them together.
The catalytic mechanism involves the binding of acyl-CoA and a long-chain alcohol to the enzyme's active site. The acyl group is transferred from CoA to the hydroxyl group of the alcohol, forming an ester bond and releasing CoA. This two-substrate reaction is characteristic of the WS/DGAT family, which uses a conserved HXXXD motif for catalysis.
Bifunctional Activity and DGAT
In simple terms: Many of these enzymes can also attach acyl groups to diacylglycerol, making them dual-purpose.
In Euglena gracilis and some bacteria, the same enzyme can catalyze both wax ester synthesis and diacylglycerol acyltransferase (DGAT) reactions, channeling acyl groups into either wax esters or triacylglycerols. This bifunctionality is a key feature of the WS/DGAT family and influences lipid storage pathways.
Substrate Specificity and Chain Length
In simple terms: Different versions of the enzyme prefer different alcohol lengths, from short to long.
Bacterial wax ester synthases exhibit diverse substrate specificities. For example, some enzymes from Marinobacter aquaeolei VT8 prefer short-chain alcohols, while others favor long-chain alcohols. Altering small and medium alcohol selectivity has been achieved through protein engineering. This specificity determines the type of wax ester produced and is crucial for industrial applications.
Physiological Roles in Wax Ester Biosynthesis
In simple terms: The enzyme makes wax esters that serve as storage or protective materials.
In jojoba, the enzyme produces high levels of liquid wax in seeds, which serves as an energy reserve. In Arabidopsis, WSD1 is involved in cuticular wax biosynthesis, and its overexpression increases wax deposition and osmotic stress tolerance. In Euglena, wax ester synthase isoenzymes play a pivotal role in wax ester biosynthesis under anaerobic conditions.
Key Genes Involved in GO:0047196 long-chain-alcohol O-fatty-acyltransferase activity
The following genes and their encoded proteins are key representatives of long-chain-alcohol O-fatty-acyltransferase activity across different organisms.
| Gene | Major Role | Research Relevance |
|---|---|---|
| WSD1 (Arabidopsis thaliana) | Wax ester synthase involved in cuticular wax biosynthesis | Overexpression improves osmotic stress tolerance |
| WS/DGAT (Marinobacter aquaeolei VT8) | Bifunctional wax ester synthase/diacylglycerol acyltransferase | Model for substrate specificity and engineering |
| wax synthase (Simmondsia chinensis) | Produces liquid wax in seeds | First plant wax synthase purified and cloned |
| WS (Euglena gracilis) | Wax ester biosynthesis under anaerobic conditions | Isoenzymes pivotal for wax ester production |
| Metagenome-derived WS | Novel wax ester synthase from soil | Demonstrates diversity and potential for biotechnology |
| Bacterial WS variants | Diverse substrate specificities | Used to study alcohol chain-length preference |
| Engineered WS mutants | Altered small/medium alcohol selectivity | Protein engineering for tailored wax esters |
| WS/DGAT from other bacteria | Lipid storage and wax ester production | Comparative studies of five bacterial enzymes |
| WSD1 homologs in Camelina sativa | Cuticular wax deposition | Improves stress tolerance in oilseed crops |
| AtWSD1 | Wax ester synthase in Arabidopsis | Model for plant cuticle engineering |
| EgWS | Euglena wax synthase | Anaerobic wax ester production |
| MaWS | Marinobacter aquaeolei wax synthase | Fifth WS/DGAT enzyme characterized |
| Jojoba WS | Seed wax production | High-level wax in transgenic Arabidopsis |
| Metagenomic WS | Soil metagenome-derived | Novel biocatalyst discovery |
| WS/DGAT mutants | Altered substrate specificity | Directed evolution studies |
| Bacterial WS1-5 | Five distinct enzymes | Substrate specificity differences |
How Is long-chain-alcohol O-fatty-acyltransferase activity Regulated?
The expression and activity of wax ester synthases are regulated at multiple levels. In plants, WSD1 expression is induced by osmotic stress, and its overexpression leads to increased wax deposition. In Euglena, wax ester synthase isoenzymes are differentially expressed under anaerobic conditions, promoting wax ester accumulation. Bacterial WS/DGAT activity can be influenced by growth conditions and carbon source, but detailed regulatory mechanisms remain less characterized.
long-chain-alcohol O-fatty-acyltransferase activity and Human Disease
| Gene | Disease / Biology | Potential Experimental Model |
|---|---|---|
| WSD1 | Osmotic stress tolerance in plants | Arabidopsis overexpression lines |
| WS/DGAT | Lipid storage and obesity-related pathways | Mouse models with heterologous expression |
| Bacterial WS | Biofilm formation and persistence | Bacterial knockout mutants |
| Euglena WS | Anaerobic wax ester production | Euglena gracilis cultures |
| Jojoba WS | Seed oil composition | Transgenic Arabidopsis |
Metabolic Disorders and Lipid Storage
Dysregulation of wax ester synthase activity could affect lipid storage and contribute to metabolic disorders. The bifunctional WS/DGAT enzymes are related to DGAT, which is implicated in obesity and insulin resistance. However, direct links to human disease are not well established, as these enzymes are primarily found in plants and microbes.
Plant Stress and Crop Resilience
In plants, wax ester synthase activity is crucial for cuticle formation, and its manipulation can improve drought and osmotic stress tolerance. Overexpression of WSD1 in Arabidopsis and Camelina enhances wax deposition and stress tolerance, suggesting agricultural applications.
Infectious Disease and Bacterial Pathogenesis
Some bacterial pathogens utilize wax ester synthases for lipid storage, which may contribute to survival in host environments. However, specific roles in pathogenesis are not well defined.
From long-chain-alcohol O-fatty-acyltransferase activity-Related Genes to Experimental Models
| Research Question | Suitable Model |
|---|---|
| Does knockout of WSD1 reduce cuticular wax? | Arabidopsis wsd1 knockout |
| Can point mutations alter substrate specificity? | Site-directed mutagenesis of WS/DGAT |
| Does knock-in of jojoba WS increase wax in seeds? | Transgenic Arabidopsis with jojoba WS |
| Where is the enzyme localized? | Tagged knock-in with GFP |
| Does overexpression improve stress tolerance? | Camelina sativa overexpressing WSD1 |
| Can bacterial WS produce novel wax esters? | E. coli expressing metagenomic WS |
How to Study the long-chain-alcohol O-fatty-acyltransferase activity Process
| Method | What It Measures | Typical Application |
|---|---|---|
| GC-MS | Wax ester composition and quantity | Profiling lipid extracts |
| In vitro enzyme assay | Catalytic activity and kinetics | Substrate specificity studies |
| Heterologous expression | Functional activity in surrogate host | Screening metagenomic libraries |
| Site-directed mutagenesis | Effect of specific residues on activity | Mechanistic studies |
| Plant transformation | In planta function and stress response | Crop improvement |
| RNA-seq | Gene expression under different conditions | Regulatory studies |
| Lipid droplet imaging | Localization of wax esters | Cellular studies |
Enzymatic Assays
In vitro assays using radiolabeled or fluorescent acyl-CoA and alcohols are standard to measure wax ester synthase activity. These assays can determine substrate specificity and kinetic parameters.
Lipidomics and GC-MS
Gas chromatography-mass spectrometry (GC-MS) is used to quantify wax ester production in cells or tissues, allowing profiling of chain lengths and saturation.
Heterologous Expression
Expressing candidate genes in E. coli or yeast enables functional characterization and screening of mutant libraries.
Plant Transformation and Stress Tests
Overexpression or knockout in Arabidopsis and Camelina followed by osmotic stress assays reveals physiological roles.
How CRISPR Can Be Used to Study GO:0047196 long-chain-alcohol O-fatty-acyltransferase activity
Knockout
CRISPR knockout of WSD1 or other wax ester synthase genes can abolish wax ester production, revealing their essential roles in cuticle formation and stress tolerance.
Point Mutation
Introducing point mutations in the catalytic HXXXD motif or substrate-binding pocket can alter enzyme activity and specificity, helping to map functional residues.
Knock-in
Knock-in of tagged or orthologous wax ester synthase genes allows tracking of protein localization and function in vivo, or transfer of wax production traits to new species.
Overexpression
CRISPR activation or transgenic overexpression of wax ester synthases can boost wax ester yields for industrial or agricultural purposes.
How EDITGENE Supports long-chain-alcohol O-fatty-acyltransferase activity Research
Researchers studying long-chain-alcohol O-fatty-acyltransferase activity-related genes often need to determine whether a candidate gene is causally involved in wax ester biosynthesis, stress tolerance, or lipid storage. EDITGENE provides comprehensive CRISPR-based services to create precise cell and animal models for functional validation.
Contact EDITGENE today to design your custom CRISPR model for long-chain-alcohol O-fatty-acyltransferase activity research.
Frequently Asked Questions About long-chain-alcohol O-fatty-acyltransferase activity
What is long-chain-alcohol O-fatty-acyltransferase activity?
It is the enzymatic activity that joins a long-chain alcohol with an acyl-CoA to form a wax ester and CoA, encoded by GO:0047196.
What genes are involved in long-chain-alcohol O-fatty-acyltransferase activity?
Key genes include WSD1 in Arabidopsis, WS/DGAT in Marinobacter, wax synthase in jojoba, and WS in Euglena.
What is another name for long-chain-alcohol O-fatty-acyltransferase?
It is commonly called wax ester synthase or wax synthase.
Which organisms have wax ester synthase?
Bacteria, plants (e.g., jojoba, Arabidopsis), and protists (e.g., Euglena) possess this activity.
How is wax ester synthase activity measured?
In vitro assays with acyl-CoA and alcohols, or GC-MS quantification of wax esters.
Can wax ester synthase be used for biofuel production?
Yes, it is used in metabolic engineering to produce wax esters and alkanes as biofuels.
What is the role of WSD1 in plants?
WSD1 is involved in cuticular wax biosynthesis and overexpression improves osmotic stress tolerance.
Are there bifunctional wax ester synthases?
Yes, many WS/DGAT enzymes also have diacylglycerol acyltransferase activity.
How can CRISPR be used to study wax ester synthase?
CRISPR knockout, knock-in, and point mutations can reveal gene function and engineer wax production.
What is the substrate specificity of wax ester synthases?
Specificity varies; some prefer short-chain alcohols, others long-chain, and can be altered by mutation.
Conclusion
GO:0047196 long-chain-alcohol O-fatty-acyltransferase activity is a fundamental enzymatic function in lipid metabolism, responsible for wax ester biosynthesis across diverse organisms. Its study offers insights into plant stress tolerance, microbial lipid storage, and industrial biotechnology. With CRISPR-based tools, researchers can precisely manipulate these genes to uncover mechanisms and engineer improved traits.
References
- 1. Choi YJ et al.. 2013. Microbial production of short-chain alkanes.. Nature 502(7472):571-4 PMID: 24077097
- 2. Vollheyde K et al.. 2020. The Fifth WS/DGAT Enzyme of the Bacterium Marinobacter aquaeolei VT8.. Lipids 55(5):479-494 PMID: 32434279
- 3. Abdullah HM et al.. 2021. Increased Cuticle Waxes by Overexpression of WSD1 Improves Osmotic Stress Tolerance in Arabidopsis thaliana and Camelina sativa.. Int J Mol Sci 22(10) PMID: 34068347
- 4. Kim NH et al.. 2016. Characterization of a Soil Metagenome-Derived Gene Encoding Wax Ester Synthase.. J Microbiol Biotechnol 26(2):248-54 PMID: 26528538
- 5. Tomiyama T et al.. 2017. Wax Ester Synthase/Diacylglycerol Acyltransferase Isoenzymes Play a Pivotal Role in Wax Ester Biosynthesis in Euglena gracilis.. Sci Rep 7(1):13504 PMID: 29044218
- 6. Barney BM et al.. 2015. Altering small and medium alcohol selectivity in the wax ester synthase.. Appl Microbiol Biotechnol 99(22):9675-84 PMID: 26205519
- 7. Barney BM et al.. 2012. Differences in substrate specificities of five bacterial wax ester synthases.. Appl Environ Microbiol 78(16):5734-45 PMID: 22685145
- 8. Lardizabal KD et al.. 2000. Purification of a jojoba embryo wax synthase, cloning of its cDNA, and production of high levels of wax in seeds of transgenic arabidopsis.. Plant Physiol 122(3):645-55 PMID: 10712527
