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Literature summary for 4.2.3.46 extracted from
- Cloning and functional expression of an (E,E)-alpha-farnesene synthase cDNA from peel tissue of apple fruit (2004), Planta, 219, 84-94.
Application
| Application | Comment | Organism |
|---|---|---|
| agriculture | Because diphenylamine treatment leaves unwanted chemical residues on the fruit, restricts export markets, and creates environmental concerns, a long-range molecular genetic strategy for control of scald by reduction of (E,E)-alpha-farnesene synthesis in scald-susceptible apples is searched. The success of this strategy will rely on our ability to identify, clone, and characterize key genes involved in alpha-farnesene biosynthesis and its regulation by ethylene. | Malus domestica |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| After screening a cDNA library generated from the peel tissue mRNA, a full-length terpene synthase cDNA 1931 nucleotides long is isolated (hot phenol RNA extraction protocol is used). The 1728-bp open reading frame encodes the 576 amino acid protein. Expression of the apple gene in Escherichia coli. | Malus domestica |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| 1-methylcyclopropene | after treatment of fruits at harvest with a blocker of ethylene action, AFS1 mRNA declines sharply over the initial 4 weeks of cold storage, and falls to nearly undetectable levels by 8 weeks | Malus domestica |  |
Localization
| Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|
| cytosol | The encoded protein appears to lack approximately 20 amino acids that are present in the N-terminal chloroplast-targeting region of monoterpene synthases. Thus, the product of this TS gene is most likely localized in the cytosol. | Malus domestica | 5829 | - |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| K+ | 500 mM are included in monoterpene synthase assay | Malus domestica | |
| Mg2+ | 10 mM are included in sesquiterpene synthase assay | Malus domestica | |
| Mn2+ | 1 mM is included in monoterpene synthase assay | Malus domestica | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 66000 | - |
without myc-epitope tag, determined by PAGE analysis | Malus domestica |
| 68000 | - |
with myc-epitope tag, determined by PAGE analysis | Malus domestica |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| farnesyl diphosphate | Malus domestica | sesquiterpene synthase activity | (E,E)-alpha-farnesene + diphosphate | - |
? | |
| farnesyl diphosphate | Malus domestica | isomers of farnesene produced in apple fruit are (E,E)-alpha and (Z,E)-alpha are in a ratio of 300:1 | (Z,E)-alpha-farnesene + diphosphate | trace amounts | ? | |
| geranyl diphosphate | Malus domestica | in monoterpene synthase assays, only (E)-beta-ocimene is produced at much reduced levels | (E)-beta-ocimene + diphosphate | - |
? | |
| geranyl diphosphate | Malus domestica | monoterpene synthase activity | (Z)-beta-ocimene + diphosphate | - |
? | |
| additional information | Malus domestica | other sesquiterpenes identified in trace amounts are (E)-nerolidol and beta-farnesene | ? | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Malus domestica | Q84LB2 | The deduced amino acid sequence of this enzyme shares the greatest degree of homology with a variety of monoterpene synthases, including alpha-(-)beta-pinene synthase, a gamma-terpinene synthase, and a (+)-limonene synthase from Citrus limon, which have 41-42% amino acid identity. A geraniol synthase from Cinnamomum tenuipilum and an isoprene synthase from Populus tremuloides have 42% identity to AFS1. The closest homology is with a linalool/nerolidol synthase from Fragaria ananassa (35% identity) and a (+)-delta-cadinene synthase from Gossypium hirsutum (33% identity).; Malus domestica Borkh., cultivar Law Rome | - |
Renatured (Commentary)
| Renatured (Comment) | Organism |
|---|---|
| myc-tagged and untagged AFS1 expressed protein in bacterial inclusion-body fractions is urea-denatured, purified and renatured prior to assay of enzymatic activity. | Malus domestica |
Source Tissue
| Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|
| epidermal cell | production of (E,E)-alpha-farnesene occurs in the epidermal (peel tissue) or adjacent hypodermal cell layers and the sesquiterpene can accumulate to high levels in the natural epicuticular coating of scald-susceptible apples during the first weeks of storage | Malus domestica | - |
| hypodermis | - |
Malus domestica | - |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| farnesyl diphosphate | sesquiterpene synthase activity | Malus domestica | (E,E)-alpha-farnesene + diphosphate | - |
? | |
| farnesyl diphosphate | isomers of farnesene produced in apple fruit are (E,E)-alpha and (Z,E)-alpha are in a ratio of 300:1 | Malus domestica | (Z,E)-alpha-farnesene + diphosphate | trace amounts | ? | |
| geranyl diphosphate | in monoterpene synthase assays, only (E)-beta-ocimene is produced at much reduced levels | Malus domestica | (E)-beta-ocimene + diphosphate | - |
? | |
| geranyl diphosphate | monoterpene synthase activity | Malus domestica | (Z)-beta-ocimene + diphosphate | - |
? | |
| additional information | other sesquiterpenes identified in trace amounts are (E)-nerolidol and beta-farnesene | Malus domestica | ? | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| (E,E)-alpha-farnesene synthase | - |
Malus domestica |
| AFS1 | - |
Malus domestica |
| sesquiterpene synthase | - |
Malus domestica |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 30 | - |
assay at | Malus domestica |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| additional information | - |
AFS1 transcript increases about 4fold in peel tissue of apple fruit during the first 4 weeks of storage at 0.5°C | Malus domestica |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 7.3 | - |
sesquiterpene synthase assay | Malus domestica |
| 7.5 | - |
monoterpene synthase assay | Malus domestica |
pH Range
| pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|
| 7 | 8 | maximal sesquiterpene synthase activity is observed | Malus domestica |
pH Stability
| pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|
| additional information | - |
sesquiterpene synthase activity declined below pH 7.0 and at pH 5.0-5.5, it is reduced more than 50% | Malus domestica |
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