Information on EC 2.5.1.63 - adenosyl-fluoride synthase

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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 ACCESSION NO.
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 ACCESSION NO.
COMMENTARY
LITERATURE
adenosyl group transfer
-
-
-
-
halogenation
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
fluoroacetate and fluorothreonine biosynthesis
-
SYSTEMATIC NAME
IUBMB Comments
S-adenosyl-L-methionine:fluoride adenosyltransferase
-
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5'-deoxy-5'-fluoroadenosine synthase
-
-
5'-FDA synthase
-
-
5'-fluoro-5'-deoxy adenosine synthetase
-
-
5'-fluoro-5'-deoxyadenosine synthase
-
-
5-fluorodeoxyadenosine synthase
-
-
-
-
adenosyl-fluoride synthase
-
-
flA protein
-
-
fluorinase
-
-
-
-
fluorinase
F8JPG4
-
fluorinase
Streptomyces cattleya DSM46488
F8JPG4
-
-
fluorinase enzyme
-
-
fluorinase, S-adenosyl-L-methionine
-
-
-
-
fluorination enzyme
-
-
SAM fluorinase
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
438583-16-5
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
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 ACCESSION NO.
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 ACCESSION NO.
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 ACCESSION NO.
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 ACCESSION NO.
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 ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
S-adenosyl-L-homocysteine
-
competent, competitive
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
EDTA
-
1 mM increases enzyme activity by 25%
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
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]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
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
-
-
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]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.029
-
S-adenosyl-homocysteine
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
development of a coupled assay with SAM synthase and fluorinase
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.9
-
-
assay at
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
180000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
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 ACCESSION NO.
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 ACCESSION NO.
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 ACCESSION NO.
LITERATURE
nickel-affinity column
-
recombinant His-tagged enzyme from Escherichia coli by nickel affinity chromatography and gel filtration
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
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 ACCESSION NO.
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 ACCESSION NO.
COMMENTARY
LITERATURE
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
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
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
-
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
-
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
-
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 ATCC 35852
-
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; 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 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
-