Information on EC 2.5.1.63 - adenosyl-fluoride synthase

New: Word Map on EC 2.5.1.63
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


The expected taxonomic range for this enzyme is: Streptomyces cattleya

EC NUMBER
COMMENTARY
2.5.1.63
-
RECOMMENDED NAME
GeneOntology No.
adenosyl-fluoride synthase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
S-adenosyl-L-methionine + fluoride = 5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
-
-
-
S-adenosyl-L-methionine + fluoride = 5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
reaction mechanism, inversion of configuration consistent with a SN2 reaction mechanism
-
S-adenosyl-L-methionine + fluoride = 5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
substrate binding structure involving F156, S158, Y177, and T80, nucleophilic substitution reaction mechanism
-
S-adenosyl-L-methionine + fluoride = 5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
C-F bond formation, the enzyme catalyses an SN2 type reaction mechanism
-
S-adenosyl-L-methionine + fluoride = 5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
reaction mechanism, overview
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
adenosyl group transfer
-
-
-
-
halogenation
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
fluoroacetate and fluorothreonine biosynthesis
-
-
SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:fluoride adenosyltransferase
-
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5-fluorodeoxyadenosine synthase
-
-
-
-
fluorinase
-
-
-
-
fluorinase, S-adenosyl-L-methionine
-
-
-
-
SAM fluorinase
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
438583-16-5
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Streptomyces cattleya ATCC 35852
-
UniProt
Manually annotated by BRENDA team
Streptomyces cattleya DSM46488
-
UniProt
Manually annotated by BRENDA team
Streptomyces cattleya NBRC14057
-
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
-
first enzyme in the fluorometabolite pathway to produce the mammalian toxin fluoroacetate and the antibiotic 4-fluorothreonine
physiological function
-
part of 4-fluorothreonine biosynthesis
physiological function
-
enzyme is essential for fluorometabolite production. Addition of 5'-fluoro-5'-deoxyadenosine to culture broth restores fluorometabolite production in the knockout mutant after a 48 h incubation, consistent with the metabolite supplementation by-passing knockout
physiological function
Streptomyces cattleya DSM46488
-
enzyme is essential for fluorometabolite production. Addition of 5'-fluoro-5'-deoxyadenosine to culture broth restores fluorometabolite production in the knockout mutant after a 48 h incubation, consistent with the metabolite supplementation by-passing knockout
-
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2'-deoxyadenosyl-L-methionine + fluoride
2'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
-
in the reverse reaction the enzyme shows 10% activity with the 2'-deoxy compound compared to the activity with the 5'-derivative
-
r
S-adenosyl-L-methionine + chloride
5'-deoxy-5'-chloroadenosine + L-methionine
show the reaction diagram
-
-
-
-
r
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
-
-
-
r
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
involved in production of toxic fluoroacetate
-
-
-
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
highly specific for S-adenosyl-L-methionine, which is tightly bound to the enzyme at the interface between C-terminal domain of one monomer and the N-terminal domain of the neighbouring monomer
-
-
?
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
reaction mechanism analysis utilizing a stereospecifically labelled substrate (5'R)-[5'-2H]-ATP in a coupled assay, inversion of configuration consistent with a SN2 reaction mechanism
i.e. 5'-FDA, NMR product analysis
-
?
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
first committed enzymatic step on the biosynthetic pathway to the fluorometabolites fluoroacetate and 4-fluorothreonine
-
-
?
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
C-F bond formation, substrate binding structure, NMR and mass spectrometry, overview
-
-
r
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
C-F bond formation, the enzyme catalyses an SN2 type reaction mechanism
-
-
?
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
18F-tagged substrate with 3 L-amino acid oxidase to remove the byproduct L-methionine enhancing the production reaction, 37C, 100 mM Tris-HCl buffer, pH 7.5, up to 4 hours, to produce 18F-tagged product, other enzymes can be added to produce several types of nucleosides
-
-
?
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
20 mg water-absorbing polymer in ionic liquid or 50 mM Tris-HCl buffer, pH 8.0, 50 nM S-adenosyl-L-methionine, 4 mircoM KF, 37C, exteraction of product 5'-deoxy-5'-fluoroadenosine with diethyl ether
-
-
?
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
Streptomyces cattleya ATCC 35852
F8JPG4
-
-
-
?
2'-deoxyadenosyl-L-methionine + fluoride
2'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
C-F bond formation, substrate binding structure, NMR and mass spectrometry, overview
in the reverse reaction the enzyme shows 10% activity with the 2'-deoxy compound compared to the activity with the 5'-derivative
-
r
additional information
?
-
-
fluoroacetate and 4-fluorothreonine accumulate in the fermentation media of Streptomyces cattleya at the mM level, when a fluoride source is added to the growth medium
-
-
-
additional information
?
-
-
fluorinase coupled enzyme systems for the synthesis of various 18F-labelled compounds, overview
-
-
-
additional information
?
-
-
substrate specificity, the enzyme does not require a planar ribose conformation of the substrate to catalyse C-F bond formation, overview, the enzyme catalyzes also transhalogenation from 2'-deoxy-5'-chloroadenosine to 2'-deoxy-fluoroadenosine via 2'-deoxy-SeAM, overview
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
-
-
-
?
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
-
-
-
r
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
involved in production of toxic fluoroacetate
-
-
-
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
first committed enzymatic step on the biosynthetic pathway to the fluorometabolites fluoroacetate and 4-fluorothreonine
-
-
?
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
18F-tagged substrate with 3 L-amino acid oxidase to remove the byproduct L-methionine enhancing the production reaction, 37C, 100 mM Tris-HCl buffer, pH 7.5, up to 4 hours, to produce 18F-tagged product, other enzymes can be added to produce several types of nucleosides
-
-
?
S-adenosyl-L-methionine + fluoride
5'-deoxy-5'-fluoroadenosine + L-methionine
show the reaction diagram
-
20 mg water-absorbing polymer in ionic liquid or 50 mM Tris-HCl buffer, pH 8.0, 50 nM S-adenosyl-L-methionine, 4 mircoM KF, 37C, exteraction of product 5'-deoxy-5'-fluoroadenosine with diethyl ether
-
-
?
additional information
?
-
-
fluoroacetate and 4-fluorothreonine accumulate in the fermentation media of Streptomyces cattleya at the mM level, when a fluoride source is added to the growth medium
-
-
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
K+
-
in the coupled assay with SAM synthase and fluorinase
Mg2+
-
in the coupled assay with SAM synthase and fluorinase
additional information
-
Mg2+ at 1 mM does not affect enzyme activity
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
S-adenosyl-L-homocysteine
-
competent, competitive
sinefungin
-
weak
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
EDTA
-
1 mM increases enzyme activity by 25%
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1.36
fluoride
-
mutant enzyme S158G, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
5.4
fluoride
-
mutant enzyme S158A, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
8.56
fluoride
-
-
10.2
fluoride
-
wild type enzyme, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
18.4
fluoride
-
mutant enzyme T80S, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
36.8
fluoride
-
mutant enzyme T80A, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
0.0008
S-adenosyl-L-methionine
-
mutant enzyme S158G, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
0.003
S-adenosyl-L-methionine
-
mutant enzyme T80A, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
0.0047
S-adenosyl-L-methionine
-
mutant enzyme T80S, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
0.0065
S-adenosyl-L-methionine
-
wild type enzyme, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
0.0092
S-adenosyl-L-methionine
-
mutant enzyme S158A, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
0.42
S-adenosyl-L-methionine
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.000012
fluoride
-
mutant enzyme S158G, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
0.000013
fluoride
-
mutant enzyme S158G, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
0.000067
fluoride
-
mutant enzyme T80A, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
0.0001
fluoride
-
mutant enzyme S158A, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
0.001
fluoride
-
mutant enzyme T80S, in 20 mM sodium phosphate buffer (pH 7.8), at 25C; wild type enzyme, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
0.0012
fluoride
-
-
2
fluoride
-
-
0.000083
S-adenosyl-L-methionine
-
mutant enzyme T80A, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
0.00015
S-adenosyl-L-methionine
-
mutant enzyme S158A, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
0.001
S-adenosyl-L-methionine
-
mutant enzyme T80S, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
0.0012
S-adenosyl-L-methionine
-
wild type enzyme, in 20 mM sodium phosphate buffer (pH 7.8), at 25C
0.074
S-adenosyl-L-methionine
-
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.029
S-adenosyl-homocysteine
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
development of a coupled assay with SAM synthase and fluorinase
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.9
-
assay at
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
180000
-
gel filtration
637250
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hexamer
-
6 * 32000, SDS-PAGE and electrospray mass spectrometry
hexamer
-
6 * 34402, (alpha,beta)3, recombinant enzyme, ESI-MS mass spectrometry
hexamer
-
a dimer of trimers
additional information
-
the N-terminal domain has a central seven-stranded beta-sheet, which combines parallel and antiparallel strands sandwiched between alpha-helices, the C-terminal domain is composed of a five- and a four-stranded antiparallel beta-sheet
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant enzyme, free from tags or bound adenosine, complexed to 2'-deoxy-5'-fluoroadenosine, 4 mg/ml protein is incubated with 20 mM ligand at 25C for 4 h, followed by crystallization using vapour diffusion against a reservoir containing 30% PEG 1000, 0.1 M phosphate-citrate pH 4.5, 0.2 M Li2SO4, X-ray diffraction structure determination and analysis at 2.4 A resolution, molecular replacement
-
purified recombinant wild-type and selenomethionine labelled enzyme with bound S-adenosyl-L-methionine, X-ray diffraction structure determination and analysis at 1.9 A resolution, structure modelling of monomers a and b
-
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
water stabilizes the enzyme, thus water-absorbing polymer and ionic liquid is used to immobilize the enzyme and keep it surrounded by water
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
nickel-affinity column
-
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography and gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli BL21; expression in Escherichia coli BL21(DE3)
-
expression of His-tagged enzyme in Escherichia coli BL21(DE3) pLysS with plasmid pET28b+
-
gene flA, DNA sequence determination and analysis, overexpression of His-tagged enzyme in Escherichia coli strain BL21(DE3) and in strain B834(DE3), the latter results in the selenomethionine labeled enzyme
-
mutant enzymes expressed in Escherichia coli C43 (DE3) cells
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D16A
-
inactive
D16N
-
3% activity compared to the wild type enzyme
D16S
-
inactive
F156A
-
3% activity compared to the wild type enzyme
F156V
-
25% activity compared to the wild type enzyme
S158A
-
38% activity compared to the wild type enzyme
S158G
-
8% activity compared to the wild type enzyme
T80A
-
15% activity compared to the wild type enzyme
T80S
-
95% activity compared to the wild type enzyme
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
analysis
-
the enzyme is applied as a catalyst for the efficient incorporation of [18F]-fluoride into [18F]-59-fluoro-59-deoxyadenosine, [18F]-59-fluoro-59-deoxyinosine and [18F]-5-fluoro-5-deoxyribose for positron emission tomography, PET, applications, useful for imaging tumors, monitoring the distribution of drugs and identifying cell and receptor degeneration in the brain
analysis
-
the fluorinase enzyme from Streptomyces cattleya is applied as a catalyst for the efficient incorporation of [18F]-fluoride into [18F]-59-fluoro-59-deoxyadenosine, [18F]-59-fluoro-59-deoxyinosine and [18F]-5-fluoro-5-deoxyribose for positron emission tomography, PET, applications, useful for imaging tumors, monitoring the distribution of drugs and identifying cell and receptor degeneration in the brain, coupled enzyme system, overview
biotechnology
-
fluorochemical production (18F-labelled organic compounds) for medicinal chemistry research (positron emission tomography), when enzyme acts together with other enzymes in the fluorometabolite production (purine nucleoside phosphorylase, isomerase, aldolase, enzyme generating fluoroacetaldehyde in a retro-aldol reaction, fluoroacetaldehyde dehydrogenase or pyridoxal phosphate-dependent enzyme) to generate fluoroacetaldehyde and 4-fluorothreonine
biotechnology
-
production of radiolabelled nucleosides as tracers for cancer cell uptake studies via positron emission tomography
biotechnology
-
the rate of the enzymatic fluorination reaction can be enhanced by using ionic liquids and immobilizing the enzyme with a water-absorbing polymer which stabilizes the enzyme, 1-octyl-3-methylimidazolium hexafluorophosphate and 1-hexyl-3-methylimidazolium hexafluorophosphate raise the conversion yield 2.4times or 1.6times compared to Tris-HCl buffer, pH 8.0, 1-hexyl-3-methylimidazolium tetrafluoroborate, 1-octyl-3-methylimidazolium tetrafluoroborate, 1-octyl-3-methylimidazolium hexafluorophosphate, and 1-hexyl-3-methylimidazolium hexafluorophosphate increase total conversation yields 2.2times to 4times more in immobilized compared to non-immobilized enzyme
synthesis
-
the enzyme is used in biotransformation reaction for production of fluorinated compounds, biotransformation protocols for coupled reaction systems, overview
synthesis
-
induced production of fluorosalinosporamide by replacing the chlorinase gene salL from Salinispora tropica with the fluorinase gene flA. Maximum yields of 4 mg/l fluorosalinosporamide production are detected at pH 6.0. The fluorosalinosporamide production yields are comparable to those attained through mutasynthesis. The major fluorinated component in the Salinispora tropica salL- flA+ extract is fluorosalinosporamide
synthesis
-
one-pot three-step continuous enzymatic synthesis of 5-fluoro-5-deoxy-D-ribose from ATP and L-methionine using S-adenosyl-L-methionine synthase, fluorinase and methylthioadenosine nucleosidase in the presence of fluoride ions. The conversion yield is 22.6% of 5-fluoro-5-deoxy-D-ribose from ATP, while the fluoride ions are generated from BF4 ionic liquids and/or the biodegradation of benzotrifluoride in the synthetic process
synthesis
-
preparation of sodium [18F]-fluoroacetate by generation of 5'-[18F]-fluoro-5'-deoxyadenosine by a fluorinase catalysed reaction of S-adenosyl-L-methionine with no carrier added [18F]-fluoride and then oxidation to [18F]-fluoroacetate by a Kuhn-Roth oxidative degradation. Na [18F]-fluoroacetate can be synthesized in 96% radiochemical purity
synthesis
Streptomyces cattleya ATCC 35852
-
preparation of sodium [18F]-fluoroacetate by generation of 5'-[18F]-fluoro-5'-deoxyadenosine by a fluorinase catalysed reaction of S-adenosyl-L-methionine with no carrier added [18F]-fluoride and then oxidation to [18F]-fluoroacetate by a Kuhn-Roth oxidative degradation. Na [18F]-fluoroacetate can be synthesized in 96% radiochemical purity, induced production of fluorosalinosporamide by replacing the chlorinase gene salL from Salinispora tropica with the fluorinase gene flA. Maximum yields of 4 mg/l fluorosalinosporamide production are detected at pH 6.0. The fluorosalinosporamide production yields are comparable to those attained through mutasynthesis. The major fluorinated component in the Salinispora tropica salL- flA+ extract is fluorosalinosporamide
-
synthesis
Streptomyces cattleya NBRC14057
-
one-pot three-step continuous enzymatic synthesis of 5-fluoro-5-deoxy-D-ribose from ATP and L-methionine using S-adenosyl-L-methionine synthase, fluorinase and methylthioadenosine nucleosidase in the presence of fluoride ions. The conversion yield is 22.6% of 5-fluoro-5-deoxy-D-ribose from ATP, while the fluoride ions are generated from BF4 ionic liquids and/or the biodegradation of benzotrifluoride in the synthetic process
-