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Information on EC 4.2.3.46 - alpha-farnesene synthase and Organism(s) Malus domestica and UniProt Accession Q84LB2
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4.2.3.46 alpha-farnesene synthaseSpecify your search results
Word Map
The taxonomic range for the selected organisms is: Malus domestica
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
mdafs1, tps03, tps02, alpha-farnesene synthase, afs-1, (e,e)-alpha-farnesene synthase, e,e-alpha-farnesene synthase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
PATHWAY SOURCE
PATHWAYS
-
-
SYSTEMATIC NAME
IUBMB Comments
(2E,6E)-farnesyl-diphosphate lyase [(3E,6E)-alpha-farnesene-forming]
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
(2E,6E)-farnesyl diphosphate
(3E,6E)-alpha-farnesene + beta-farnesene + diphosphate
when farnesyl diphosphate, synthesised from 96% (E,E)-farnesol, is incubated with recombinant protein, (E,E)- and (Z,E)-alpha-farnesene are produced in a ratio of 96:4, respectively
-
-
?
farnesyl diphosphate
(E,E)-alpha-farnesene + diphosphate
sesquiterpene synthase activity
-
-
?
farnesyl diphosphate
(Z,E)-alpha-farnesene + diphosphate
isomers of farnesene produced in apple fruit are (E,E)-alpha and (Z,E)-alpha are in a ratio of 300:1
trace amounts
-
?
geranyl diphosphate
(E)-beta-ocimene + diphosphate
in monoterpene synthase assays, only (E)-beta-ocimene is produced at much reduced levels
-
-
?
geranyl diphosphate
(Z)-beta-ocimene + diphosphate
monoterpene synthase activity
-
-
?
geranyl diphosphate
linalool + (Z)-beta-ocimene + (E)-beta-ocimene + beta-myrcene
at 18% of the optimised rate for alpha-farnesene synthesis from farnesyl diphosphate
-
-
?
(2E,6E)-farnesyl diphosphate
(3E,6E)-alpha-farnesene + diphosphate
-
-
-
?
geranyl diphosphate
(E)-beta-ocimene + beta-myrcene
poor substrate
-
-
?
(2E,6E)-farnesyl diphosphate
(3E,6E)-alpha-farnesene + diphosphate
-
-
-
?
(2E,6E)-farnesyl diphosphate
(3E,6E)-alpha-farnesene + diphosphate
commentary
-
-
?
(2E,6E)-farnesyl diphosphate
(3E,6E)-alpha-farnesene + diphosphate
alpha-farnesene synthase is a key enzyme in the pathway of alpha-farnesene synthesis
-
-
?
(2E,6E)-farnesyl diphosphate
(3E,6E)-alpha-farnesene + diphosphate
apha-farnesene synthase is a key enzyme in the pathway of alpha-farnesene synthesis
-
-
?
additional information
?
-
other sesquiterpenes identified in trace amounts are (E)-nerolidol and beta-farnesene
-
-
?
additional information
?
-
the monoterpene synthase ((E)-beta-ocimene and beta-myrcene) and sesquiterpene synthase (alpha-farnesene) products produced by the mutated and wild-type enzymes are identical, there are no significant alterations in the ratios of the alpha-farnesene isomers produced
-
-
?
additional information
?
-
the monoterpene synthase ((E)-beta-ocimene and beta-myrcene) and sesquiterpene synthase (alpha-farnesene) products produced by the mutated and wild-type enzymes are identical, there are no significant alterations in the ratios of the alpha-farnesene isomers produced
-
-
?
additional information
?
-
-
the monoterpene synthase ((E)-beta-ocimene and beta-myrcene) and sesquiterpene synthase (alpha-farnesene) products produced by the mutated and wild-type enzymes are identical, there are no significant alterations in the ratios of the alpha-farnesene isomers produced
-
-
?
additional information
?
-
although (E,E)-farnesyl diphosphate is the preferred substrate, the enzyme accepts all four isomeric forms of the farnesyl diphosphate precursor. Both isomers of beta-farnesene are also synthesised by the enzyme presumably from a specific farnesene isomer. The enzyme also produces alpha-farnesene by a reaction involving coupling of geranyl diphosphate and isoprenyl diphosphate but at less than 1% of the rate with farnesyl diphosphate
-
-
?
additional information
?
-
-
although (E,E)-farnesyl diphosphate is the preferred substrate, the enzyme accepts all four isomeric forms of the farnesyl diphosphate precursor. Both isomers of beta-farnesene are also synthesised by the enzyme presumably from a specific farnesene isomer. The enzyme also produces alpha-farnesene by a reaction involving coupling of geranyl diphosphate and isoprenyl diphosphate but at less than 1% of the rate with farnesyl diphosphate
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
farnesyl diphosphate
(E,E)-alpha-farnesene + diphosphate
sesquiterpene synthase activity
-
-
?
farnesyl diphosphate
(Z,E)-alpha-farnesene + diphosphate
isomers of farnesene produced in apple fruit are (E,E)-alpha and (Z,E)-alpha are in a ratio of 300:1
trace amounts
-
?
geranyl diphosphate
(E)-beta-ocimene + diphosphate
in monoterpene synthase assays, only (E)-beta-ocimene is produced at much reduced levels
-
-
?
geranyl diphosphate
(Z)-beta-ocimene + diphosphate
monoterpene synthase activity
-
-
?
(2E,6E)-farnesyl diphosphate
(3E,6E)-alpha-farnesene + diphosphate
-
-
-
?
(2E,6E)-farnesyl diphosphate
(3E,6E)-alpha-farnesene + diphosphate
commentary
-
-
?
(2E,6E)-farnesyl diphosphate
(3E,6E)-alpha-farnesene + diphosphate
alpha-farnesene synthase is a key enzyme in the pathway of alpha-farnesene synthesis
-
-
?
(2E,6E)-farnesyl diphosphate
(3E,6E)-alpha-farnesene + diphosphate
apha-farnesene synthase is a key enzyme in the pathway of alpha-farnesene synthesis
-
-
?
additional information
?
-
other sesquiterpenes identified in trace amounts are (E)-nerolidol and beta-farnesene
-
-
?
additional information
?
-
the monoterpene synthase ((E)-beta-ocimene and beta-myrcene) and sesquiterpene synthase (alpha-farnesene) products produced by the mutated and wild-type enzymes are identical, there are no significant alterations in the ratios of the alpha-farnesene isomers produced
-
-
?
additional information
?
-
the monoterpene synthase ((E)-beta-ocimene and beta-myrcene) and sesquiterpene synthase (alpha-farnesene) products produced by the mutated and wild-type enzymes are identical, there are no significant alterations in the ratios of the alpha-farnesene isomers produced
-
-
?
additional information
?
-
-
the monoterpene synthase ((E)-beta-ocimene and beta-myrcene) and sesquiterpene synthase (alpha-farnesene) products produced by the mutated and wild-type enzymes are identical, there are no significant alterations in the ratios of the alpha-farnesene isomers produced
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
K+
Mg2+
Mn2+
K+
activity is dependent on K+, the potassium binding region is defined
K+
MdAFS1 retains up to 12% of its activity in the absence of K+, enzyme contains K+ binding region, MdAFS1 exhibits a type II K+ response, MdAFS1 is not absolutely dependent upon M+ its unequivocal classification as type I or type II K+ activated, or that of any other terpene synthases, will not be possibl, then type I enzymes can exhibit type II kinetics and vice versa.
K+
30-50 mM, enhances activity 5fold. Km: 3 mM. Addition of K+ reduces monoterpene synthase activity
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
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
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0026
farnesyl diphosphate
mutant S487A, 50 mM KCl, 10 mM MgCl2
0.0086
farnesyl diphosphate
mutant D484A, 50 mM KCl, 10 mM MgCl2
0.022
farnesyl diphosphate
without K+, sesquiterpene synthase activity in mutant S487K is independent of K+
0.039
farnesyl diphosphate
mutant S488A, 50 mM KCl, 10 mM MgCl2
0.0533
farnesyl diphosphate
wild-type MdAFS, 50 mM KCl, 10 mM MgCl2
0.0613
farnesyl diphosphate
mutant S485A, 50 mM KCl, 10 mM MgCl2
0.0026
(2E,6E)-farnesyl diphosphate
pH 7.5, 30°C, mutant enzyme S487A
0.0086
(2E,6E)-farnesyl diphosphate
pH 7.5, 30°C, mutant enzyme D484A
0.039
(2E,6E)-farnesyl diphosphate
pH 7.5, 30°C, mutant enzyme S488A
0.0553
(2E,6E)-farnesyl diphosphate
pH 7.5, 30°C, wild-type enzyme
0.0613
(2E,6E)-farnesyl diphosphate
pH 7.5, 30°C, mutant enzyme S485A
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
S487K mutant has 35-45% of the wild-type activity with farnesyl diphosphate with no significant alterations in the alpha-farnesene isomer ratios produced and a small decrease in catalytic efficiency (wild-type and S487K respective kcat/Km values of 17.5 and 13.3 mM/s)
additional information
S487K mutant has 35-45% of the wild-type activity with farnesyl diphosphate with no significant alterations in the alpha-farnesene isomer ratios produced and a small decrease in catalytic efficiency (wild-type and S487K respective kcat/Km values of 17.5 and 13.3 mM/s)
additional information
-
S487K mutant has 35-45% of the wild-type activity with farnesyl diphosphate with no significant alterations in the alpha-farnesene isomer ratios produced and a small decrease in catalytic efficiency (wild-type and S487K respective kcat/Km values of 17.5 and 13.3 mM/s)
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.5
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
genomic structure and sequence polymorphism of E,E-alpha farnesene synthase gene in Malus domestica; Borkh.
UniProt
genomic structure and sequence polymorphism of E,E-alpha-farnesene synthase gene in Malus domestica; Borkh.
UniProt
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
UniProt
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
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
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
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.
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). AFS1_MALDO
576
0
66183
Swiss-Prot
other Location (Reliability: 3)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D326A
alpha-farnesene synthase, monoterpene synthase and prenyltransferase activities are lost in the mutant
D326A/D330A
alpha-farnesene synthase, monoterpene synthase and prenyltransferase activities are lost in the mutant
D484A
85% loss of sesquiterpene synthase activity compared with wild-type enzyme when K+ is present
S485A
exhibits marginally increased sesquiterpene synthase activities, and an approximate 2fold increase in monoterpene synthase activity compared with the wild-type enzyme
S487A
95% decrease in sesquiterpene synthase activity compared with wild-type enzyme when K+ is present
S487K
The S487K mutant has 35-45% of the wild-type activity with farnesyl diphosphate, with no significant alterations in the alpha-farnesene isomer ratios produced and a small decrease in catalytic efficiency
S488A
mutant shows decreases in both sesqui- and monoterpene synthase activities compared with the WT enzyme, with mono-TPS activity being reduced more than sesquiterpene synthase activity. Sesquiterpene synthase (alpha-farnesene) products produced by the mutated and wild-type enzymes are identical, there are no significant alterations in the ratios of the alpha-farnesene isomers produced.
D484A
85% loss of sesquiterpene synthase activity compared with wild-type enzyme. Little change in K+-independent activity
S485A
mutant exhibits marginally increased sesquiterpene synthase activities, and an approximate 2fold increase in monoterpene synthase activity compared with the wild-type enzyme
S487A
95% decrease in sesquiterpene synthase activity compared with wild-type enzyme. Little change in K+-independent activity
S487K
mutant has 3545% of the wild-type activity with farnesyl diphosphate, with no significant alterations in the alpha-farnesene isomer ratios produced and a small decrease in catalytic efficiency
S488A
mutant shows decreases in both sesqui- and mono-terpene synthases activities, compared with the wild-type enzyme, with mono-terpene synthases activity being reduced more than sesqui-terpene synthases activity
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
sesquiterpene synthase activity declined below pH 7.0 and at pH 5.0-5.5, it is reduced more than 50%
694765
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
AFS1 transcript increases about 4fold in peel tissue of apple fruit during the first 4 weeks of storage at 0.5°C
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
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.
mutated enzymes are generated using the QuickChange II site-directed mutagenesis kit. The PCR-based mutagenesis protocol is performed using pET-30a harbouring the MdAFS1 cDNAs as template. The single-site mutant enzymes overexpressed in Escherichia coli.
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
expression of pMdAFS1 is repressed by 1-methylcyclopropene treatment, pMdAFS1 expression is controlled by ethylene
MdAFS1 transcript abundance increases rapidly in untreated Cortland and Law Rome fruit, reaching close to maximal values after 2 weeks of storage, whereas in Idared controls it increases through 4 weeks, declines slightly, and then increases to a maximum by 10 weeks. 1-Methylcyclopropene treatment initially suppresses MdAFS1 expression in all three cultivars. However, Cortland and Law Rome escape from this suppression after 10-15 weeks, with MdAFS1 transcript abundance increasing to maximal control levels by 15-20 weeks. In Cortland, but not in Law Rome, this delayed increase in expression occurrs sooner in green than in red tissue. MdAFS1 transcript levels increase gradually in 1-methylcyclopropene-treated Idared fruit, but throughout storage remains much lower than the maximum levels in untreated fruit
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
myc-tagged and untagged AFS1 expressed protein in bacterial inclusion-body fractions is urea-denatured, purified and renatured prior to assay of enzymatic activity.
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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.
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Green, S.; Squire, C.J.; Nieuwenhuizen, N.J.; Baker, E.N.; Laing, W.
Defining the potassium binding region in an apple terpene synthase
J. Biol. Chem.
284
8661-8669
2009
Malus domestica (B2ZZ11), Malus domestica (Q84LB2), Malus domestica
Pechous S.W.; Whitaker, B.D.
Cloning and functional expression of an (E,E)-alpha-farnesene synthase cDNA from peel tissue of apple fruit
Planta
219
84-94
2004
Malus domestica (Q84LB2)
Ban, Y.; Oyama-Okubo, N.; Honda, C.; Nakayama, M.; Moriguchi, T.
Emitted and endogenous volatiles in 'Tsugaru' apple: The mechanism of ester and (E,E)-?-farnesene accumulation
Food Chem.
118
272-277
2010
Malus domestica (B2ZZ11)
Yuan, K.; Liu, Q.; Li, B.; Zhang, L.
Genomic structure and sequence polymorphism of E,E-alpha-farnesene synthase gene in apples (Malus domestica Borkh.)
Front. Agric. China
2
190-193
2008
Malus domestica (Q84LB2)
-
Beuning, L.; Green, S.; Yauk, Y.-K.
The genomic sequence of AFS-1 - an alpha-farnesene synthase from the apple cultivar Royal Gala
Front. Agric. China
4
74-78
2010
Malus domestica (Q84LB2)
-
Tsantili, E.; Gapper, N.E.; Arquiza, J.M.; Whitaker, B.D.; Watkins, C.B.
Ethylene and alpha-farnesene metabolism in green and red skin of three apple cultivars in response to 1-methylcyclopropene (1-MCP) treatment
J. Agric. Food Chem.
55
5267-5276
2007
Malus domestica (B2ZZ11), Malus domestica
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