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Information on EC 2.8.1.6 - biotin synthase and Organism(s) Escherichia coli and UniProt Accession P12996
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2.8.1.6 biotin synthaseIUBMB Comments
The enzyme binds a [4Fe-4S] and a [2Fe-2S] cluster. In every reaction cycle, the enzyme consumes two molecules of AdoMet. The first reaction produces 5'-deoxyadenosine and 4,5-secobiotin. Reaction with another equivalent of AdoMet results in abstraction of the C-6 methylene pro-S hydrogen atom from 4,5-secobiotin, and the resulting carbon radical is quenched via formation of an intramolecular C-S bond, thus closing the biotin tetrahydrothiophene ring. The sulfur donor is believed to be the [2Fe-2S] cluster, which is sacrificed in the process, so that in vitro the reaction is a single turnover. In vivo, the [2Fe-2S] cluster can be reassembled by the Isc or Suf iron-sulfur cluster assembly systems, to allow further catalysis.
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Word Map
- 2.8.1.6
- s-adenosylmethionine
-
iron-sulfur
- adomet
-
5\'-deoxyadenosyl
- 5'-deoxyadenosine
- sphaericus
-
thiophane
- flavodoxins
-
sulfonium
-
adomet-dependent
- lipoate
-
desulfurase
- formate-lyase
-
deoxyadenosyl
-
2,3-aminomutase
- nutrition
The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Reaction Schemes
+
+
2
+
2
=
+
+
2
+
2
+
2
+
sulfur-(sulfur carrier)
+
2
+
2
=
+
(sulfur carrier)
+
2
+
2
+
2
Synonyms
biotin synthase, biotin synthetase, ncgl0071, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
BIO2 protein (Arabidopsis thaliana clone pMP101 gene BIO2 reduced)
-
-
-
-
BioB
biotin synthase (Saccharomyces cerevisiae strain 20B-12 clone pUCH2.4 gene BIO2)
-
-
-
-
biotin synthase (Treponema pallidum gene TP0228)
-
-
-
-
biotin synthetase
-
-
-
-
biotin synthetase (Aquifex aeolicus gene bioB)
-
-
-
-
biotin synthetase (Arabidopsis thaliana clone pMB101 gene BIO2 reduced)
-
-
-
-
biotin synthetase (Bacillus subtilis gene bioB)
-
-
-
-
biotin synthetase (Chlamydia trachomatis gene birA)
-
-
-
-
GenBank AE000716-derived protein GI 2983482
-
-
-
-
GenBank AE001204-derived protein GI 3322497
-
-
-
-
GenBank AE001343-derived protein GI 3329182
-
-
-
-
GenBank AF008220-derived protein GI 2293187
-
-
-
-
GenBank U24147-derived protein
-
-
-
-
GenBank U31806-derived protein GI 1403662
-
-
-
-
GenBank U51869-derived protein GI 1277029
-
-
-
-
GenBank Z99119-derived protein GI 2635504
-
-
-
-
synthetase, biotin
-
-
-
-
synthetase, biotin (Arabidopsis thaliana clone lambdaBIO2 gene bioB)
-
-
-
-
synthetase, biotin (Arabidopsis thaliana clone pMP101 gene BIO2 reduced)
-
-
-
-
synthetase, biotin (Arabidopsis thaliana clone pYESCBS1 gene bioB)
-
-
-
-
synthetase, biotin (Bacillus subtilis clone pBIO100/pBIO350/pBIO201 gene bioB)
-
-
-
-
synthetase, biotin (Bacillus subtilis gene bioB)
-
-
-
-
synthetase, biotin (Saccharomyces cerevisiae strain 20B-12 clone pUCH2.4 gene BIO2 reduced)
-
-
-
-
BioB
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
dethiobiotin + sulfur-(sulfur carrier) + 2 S-adenosyl-L-methionine + 2 reduced [2Fe-2S] ferredoxin = biotin + (sulfur carrier) + 2 L-methionine + 2 5'-deoxyadenosine + 2 oxidized [2Fe-2S] ferredoxin
substrates are placed between the (4Fe-4S) cluster, essential for radical generation, and the (2Fe-2S) cluster
dethiobiotin + sulfur-(sulfur carrier) + 2 S-adenosyl-L-methionine + 2 reduced [2Fe-2S] ferredoxin = biotin + (sulfur carrier) + 2 L-methionine + 2 5'-deoxyadenosine + 2 oxidized [2Fe-2S] ferredoxin
dethiobiotin + sulfur-(sulfur carrier) + 2 S-adenosyl-L-methionine + 2 reduced [2Fe-2S] ferredoxin = biotin + (sulfur carrier) + 2 L-methionine + 2 5'-deoxyadenosine + 2 oxidized [2Fe-2S] ferredoxin
mechanism
-
dethiobiotin + sulfur-(sulfur carrier) + 2 S-adenosyl-L-methionine + 2 reduced [2Fe-2S] ferredoxin = biotin + (sulfur carrier) + 2 L-methionine + 2 5'-deoxyadenosine + 2 oxidized [2Fe-2S] ferredoxin
radical chemistry is involved in the mechanism of biotin synthase
-
dethiobiotin + sulfur-(sulfur carrier) + 2 S-adenosyl-L-methionine + 2 reduced [2Fe-2S] ferredoxin = biotin + (sulfur carrier) + 2 L-methionine + 2 5'-deoxyadenosine + 2 oxidized [2Fe-2S] ferredoxin
reaction proceeds via intermediate involving the (2Fe-2S) cluster, with a rate-determining step following the formation of this intermediate
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
sulfur atom transfer
sulfur atom transfer
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
dethiobiotin:sulfur-(sulfur carrier) sulfurtransferase
The enzyme binds a [4Fe-4S] and a [2Fe-2S] cluster. In every reaction cycle, the enzyme consumes two molecules of AdoMet. The first reaction produces 5'-deoxyadenosine and 4,5-secobiotin. Reaction with another equivalent of AdoMet results in abstraction of the C-6 methylene pro-S hydrogen atom from 4,5-secobiotin, and the resulting carbon radical is quenched via formation of an intramolecular C-S bond, thus closing the biotin tetrahydrothiophene ring. The sulfur donor is believed to be the [2Fe-2S] cluster, which is sacrificed in the process, so that in vitro the reaction is a single turnover. In vivo, the [2Fe-2S] cluster can be reassembled by the Isc or Suf iron-sulfur cluster assembly systems, to allow further catalysis.
CAS REGISTRY NUMBER
COMMENTARY
153554-27-9
synthetase, biotin (Saccharomyces cerevisiae strain 20B-12 clone pUCH2.4 gene BIO2 reduced) /biotin synthase (Saccharomyces cerevisiae strain 20B-12 clone pUCH2.4 gene BIO2)
174764-24-0
synthase, biotin (Arabidopsis thaliana clone lambdaBIO2 gene bioB) /BIO2 protein (Arabidopsis thaliana clone pMP101 gene BIO2 reduced) /genBank U24147-derived protein /genBank U31806-derived protein GI 1403662 /synthetase, biotin (Arabidopsis thaliana clone pMP101 gene BIO2 reduced) /synthetase, biotin (Arabidopsis thaliana clone pYESCBS1 gene bioB)
179608-56-1
synthetase, biotin (Bacillus subtilis clone pBIO100/pBIO350/pBIO201 gene bioB) /biotin synthetase (Bacillus subtilis gene bioB) /genBank AF008220-derived protein GI 2293187 /genBank U51869-derived protein GI 1277029 /genBank Z99119-derived protein GI 2635504 /Synthetase, biotin (Bacillus subtilis gene bioB)
204794-88-7
biotin synthetase (Aquifex aeolicus gene bioB) /genBank AE000716-derived protein GI 2983482
209603-31-6
biotin synthase (Treponema pallidum gene TP0228) /genBank AE001204-derived protein GI 3322497
215108-34-2
biotin synthetase (Chlamydia trachomatis gene birA) /genBank AE001343-derived protein GI 3329182
80146-93-6
-
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
9-mercaptodethiobiotin + [S] + S-adenosyl-L-methionine
biotin + ?
-
-
-
?
dethiobiotin + sulfur + S-adenosyl-L-methionine
biotin + L-methionine + 5'-deoxyadenosine
-
-
-
?
9-mercaptodethiobiotin + S-adenosyl-L-methionine
biotin + ?
-
presence of S-adenosyl-L-methionine is required
-
-
?
9-mercaptodethiobiotin + [S] + S-adenosyl-L-methionine
biotin + L-methionine + 5'-deoxyadenosine
-
-
-
-
ir
dethiobiotin + Na2Se
selenobiotin
-
enzyme depleted of iron and sulfur and reconstituted with FeCl3 and Na2Se yielding an [2Fe-2Se]2+ cluster. Activity with Na2Se is lower than that of the as-isolated enzyme with Na2S
-
-
?
dethiobiotin + sulfur + 2 S-adenosyl-L-methionine
biotin + 2 L-methionine + 2 5'-deoxyadenosine
dethiobiotin + sulfur + S-adenosyl-L-methionine
biotin + L-methionine + 5'-deoxyadenosine
-
-
-
-
?
dethiobiotin + [S] + 2 S-adenosyl-L-methionine
biotin + 2 L-methionine + 2 5'-deoxyadenosine
-
-
-
-
ir
dethiobiotin + [S] + S-adenosyl-L-methionine
9-mercaptodethiobiotin + L-methionine + 5'-deoxyadenosine
-
9-mercaptodethiobiotin is generated as a ligand to the [2Fe-2S]2+ cluster during the reaction
-
-
ir
L-cysteine
L-alanine + sulfide
-
enzyme displays cysteine desulfurase activity, providing it with the ability to mobilize sulfur from free cysteine
-
?
dethiobiotin + [S] + S-adenosyl-L-methionine
biotin + L-methionine + 5'-deoxyadenosine
-
-
-
ir
dethiobiotin + [S] + S-adenosyl-L-methionine
biotin + L-methionine + 5'-deoxyadenosine
dethiobiotin is 5-methyl-2-oxo-4-imidazolidinehexanoic acid
-
-
?
dethiobiotin + sulfur
biotin
-
-
-
-
?
dethiobiotin + sulfur
biotin
-
insertion of a sulfur atom between the inactive methyl and methylene carbon atoms adjacent to the imidazolinone ring
-
-
?
dethiobiotin + sulfur
biotin
-
insertion of a sulfur atom between the saturated C6 and C9 carbons of dethiobiotin
-
?
dethiobiotin + sulfur
biotin
-
hypothesis: sulfur of biotin is derived from the [Fe-S] center of the enzyme
-
-
?
dethiobiotin + sulfur
biotin
-
catalyzes the last step of the biosynthesis of biotin
-
-
?
dethiobiotin + sulfur
biotin
-
biotin synthase is active for only one turnover, during which the [2Fe-2S]2+ cluster is destroyed, one sulfide from the cluster is incorporated as the biotin thiophane sulfur, while Fe2+ ions and the remaining S2- ion are released from the protein
-
-
?
dethiobiotin + sulfur + 2 S-adenosyl-L-methionine
biotin + 2 L-methionine + 2 5'-deoxyadenosine
-
9-mercaptodethiobiotin is formed as a competent catalytic intermediate by Escherichia coli biotin synthase
-
-
?
dethiobiotin + sulfur + 2 S-adenosyl-L-methionine
biotin + 2 L-methionine + 2 5'-deoxyadenosine
-
The biotin synthase (BioB) reaction does not require pyridoxal phosphate in vivo. Therefore, the biotin sulfur atom cannot be derived via an intrinsic pyridoxal phosphate-dependent BioB cysteine desulfurase activity
-
-
?
dethiobiotin + [S] + S-adenosyl-L-methionine
biotin + L-methionine + 5'-deoxyadenosine
-
-
-
-
?
dethiobiotin + [S] + S-adenosyl-L-methionine
biotin + L-methionine + 5'-deoxyadenosine
-
overall reaction
-
-
ir
additional information
?
-
-
cysteine and S-adenosylmethionine are no sulfur donors
-
-
?
additional information
?
-
-
covalent alteration of enzyme during turnover suggesting a role of protein as a reagent, not a catalyst
-
-
?
additional information
?
-
-
enzyme has very modest catalytic power. 20-60 molecules of biotin are formed per molecule of enzyme, which is easily degraded. Substantial unfolding of enzyme is required to allow rebuilding of the (2Fe-2S) cluster after each turnover. Such unfolding allows restoration of the cluster, but at the cost of exposure of the protein to proteolytic attack
-
-
?
additional information
?
-
-
reaction has a turnover of one. Enzyme has no cysteine desulfurase activity, the required sulfide has to be added as Na2S. The active enzyme cannot be fully restored after one turn
-
-
?
additional information
?
-
-
enzyme depleted of iron and sulfur and reconstituted with FeCl3 and Na2Se to enable formation an [2Fe-2Se]2+ cluster. The [2Fe-2Se]2+ enzyme yields a mixture of biotin and selenobiotin in presence of Na2S
-
-
?
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
dethiobiotin + sulfur + S-adenosyl-L-methionine
biotin + L-methionine + 5'-deoxyadenosine
-
-
-
?
dethiobiotin + sulfur + 2 S-adenosyl-L-methionine
biotin + 2 L-methionine + 2 5'-deoxyadenosine
dethiobiotin + sulfur + S-adenosyl-L-methionine
biotin + L-methionine + 5'-deoxyadenosine
-
-
-
-
?
dethiobiotin + sulfur
biotin
-
hypothesis: sulfur of biotin is derived from the [Fe-S] center of the enzyme
-
-
?
dethiobiotin + sulfur
biotin
-
catalyzes the last step of the biosynthesis of biotin
-
-
?
dethiobiotin + sulfur
biotin
-
biotin synthase is active for only one turnover, during which the [2Fe-2S]2+ cluster is destroyed, one sulfide from the cluster is incorporated as the biotin thiophane sulfur, while Fe2+ ions and the remaining S2- ion are released from the protein
-
-
?
dethiobiotin + sulfur + 2 S-adenosyl-L-methionine
biotin + 2 L-methionine + 2 5'-deoxyadenosine
-
9-mercaptodethiobiotin is formed as a competent catalytic intermediate by Escherichia coli biotin synthase
-
-
?
dethiobiotin + sulfur + 2 S-adenosyl-L-methionine
biotin + 2 L-methionine + 2 5'-deoxyadenosine
-
The biotin synthase (BioB) reaction does not require pyridoxal phosphate in vivo. Therefore, the biotin sulfur atom cannot be derived via an intrinsic pyridoxal phosphate-dependent BioB cysteine desulfurase activity
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
[2Fe-2S]-center
a reaction intermediate includes Arg260 as a deprotonated guanidino group, coordinated to the ferrous iron of the auxiliary [2Fe-2S] cluster as a monodentate ligand. The 9-mercaptodethiobiotin C9 is covalently coordinated to the auxiliary [2Fe-2S] cluster, which maintains the geometric and electronic structure characteristics of a typical reduced [2Fe-2S]+ cluster, with most of the spin density localized on the cluster and only a small positive spin density residing on the 9-mercaptodethiobiotin ligand via through-bond (C9 and my-thiolate) spin delocalization. C6, the target of the second hydrogen-atom abstraction, is now located in close proximity to the newly acquired thiolate sulfur
Fe-S center
the active site of the enzyme involves a [4Fe-4S] cluster
Fe-S center
the enzyme contains one [4Fe-4S]2+ cluster and one air-stable [2Fe-2S]2+ cluster per monomer
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Fe2+
FeCl3
Iron
Na2S
FeCl3
-
sulfur of the iron-sulfur cluster is provided by cysteine desulfurase EC 2.8.1.7
Iron
-
aerobically purified enzyme contains 1.2-1.5 [2Fe-2S] clusters per monomer. Upon reduction the [Fe2-S2] clusters are converted to [Fe4-S4] clusters. The dominant stable cluster state for the enzyme is a dimer containing 2 [Fe2-S2] clusters and 2 [Fe4-S4] clusters
Iron
-
1 [2Fe-2S] cluster per monomer, but enzyme can be reconstituted to contain an additional [4Fe-4S] cluster, both clusters must be present for tight substrate binding
Iron
-
1 [2Fe-2S] and 1 [4Fe-4S] cluster per monomer are essential for optimal activity
Iron
-
1 [2Fe-2S] per monomer, but enzyme is more active when reconstituted with an additional [4Fe-4S] cluster
Iron
-
in a partially purified fraction the presence of a S2- source and Fe2+ converts the predominant [2Fe-2S] into a 1:1 mixture of [2Fe-2S] and [4Fe-4S], reduced [4Fe-4S] is involved in mediating the cleavage of S-adenosylmethionine and reduced [2Fe-2S] is the sulfur source of biotin
Iron
-
the 104000 Da dimeric enzyme form contains a single [2Fe-2S] cluster per dimer
Iron
-
the 82000 Da dimeric enzyme form contains one [2Fe-2S]cluster per monomer
Iron
-
enzyme contains a (4FE-4S) cluster that is stable during the reaction and bound to S-adenosyl-L-methionine. Additionally, enzyme contains a (2Fe-2S) cluster. About 2/3 of the (2Fe-2S) clusters are degraded by the end of a turnover experiment, degradation is initiated by reduction of the cluster
Iron
-
enzyme contains two distinct Fe-S cluster binding sites, one site accomodates a (2Fe-2S)2+ cluster with partial noncysteinyl ligation, the other site accomodates a (4Fe-4S)2+ cluster that binds S-adenosyl-L-methionine and undergoes O2-induced degradation
Iron
-
in as-prepared sample, present as (2Fe-2s)2+ cluster with incomplete cysteinyl-S coordination, reversible conversion by dithionite yields (4Fe-4S)2+, Mossbauer studies
Iron
-
the as-isolated form of enzyme contains an air-stable [2Fe-2S]2+ center. Enzyme can additionally accomodate an air-sensitive [4Fe-4S]2+ center which is generated by incubation under anaerobic conditions with Fe2+ and S2-. With respect to iron cluster content and characteristics, mutants N151A, H152A, N153A, D155A are similar to wild-type
Iron
-
[2Fe-2S]2+ cluster coordinated by C97, C128, C188 and R260. Wild-type enzyme contains 1.6 iron atoms per monomer
Iron
-
BioB appears to be resistant to degradation and capable of multiple turnovers only under high-iron conditions that favor repair of the FeS clusters, a process most likely mediated by the Isc or Suf iron-sulfur cluster assembly systems.
Iron
-
loss of the FeS clusters results in decreased thermal stability and apparent localized unfolding of BioB, particularly in the regions around Arg168 and Arg245, but not global unfolding
Iron
-
the [2Fe-2S]2+ cluster is both the sulfur-donating substrate and the sulfur-oxidizing cofactor
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
L-methionine
-
product inhibition. The combination of both products in equimolar concentrations
results in more than simple additive inhibition, suggesting that they bind cooperatively to the enzyme.
5'-deoxyadenosine
-
strong inhibitor, reversible, 90% inhibition upon addition of 1 equivalent of 5'-deoxyadenosine with regard to the enzyme
5'-deoxyadenosine
-
product inhibition. The combination of both products in equimolar concentrations
results in more than simple additive inhibition, suggesting that they bind cooperatively to the enzyme.
additional information
-
not inhibitory: pyridoxal 5-phosphate, deoxyadenosine
-
additional information
-
5'-methylthioadenosine is not an inhibitor. Neither biotin nor iminobiotin is a significant inhibitor
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
pyridoxal 5'-phosphate
-
required for cysteine desulfurase activity of the enzyme
additional information
-
a labile low-molecular-weight product of the 7,8-diaminoperlargonic acid aminotransferase reaction stimulates by 3fold
-
dithiothreitol
-
10fold stimulation of cysteine desulfurase activity of the enzyme at 20 mM
S-adenosyl-L-methionine
-
two molecules of S-adenosyl-L-methionine are used to synthesize one molecule of biotin, one from dethiobiotin to the intermediate, and a second from the intermediate to biotin
S-adenosyl-L-methionine
-
one molecule S-adenosyl-L-methionine is required to form one molecule of biotin
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.2
9-Mercaptodethiobiotin
pH and temperature not specified in the publication
0.005
dethiobiotin
pH and temperature not specified in the publication
0.01
S-adenosyl-L-methionine
-
in 50 mM Tris-HCl, 100 mM KCl, and 5 mM dithiothreitol, pH 8.0, at 37°C
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
BioB is active for only one turnover, during which the [2Fe-2S]2+ cluster is destroyed, one sulfide from the cluster is incorporated as the biotin thiophane sulfur, while Fe2+ ions and the remaining S2- ion are released from the protein
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.00065
S-adenosyl-L-homocysteine
-
in 50 mM Tris-HCl, 100 mM KCl, and 5 mM dithiothreitol, pH 8.0, at 37°C
0.075
sinefungin
-
in 50 mM Tris-HCl, 100 mM KCl, and 5 mM dithiothreitol, pH 8.0, at 37°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
metabolism
biotin synthase catalyzes the final step in the biosynthesis of biotin
physiological function
a semistable intermediate in the formation of the first C-S bond is 9-mercaptodethiobiotin linked to a paramagnetic [2Fe-2S] cluster through one of its bridging sulfides. C6, the target of the second hydrogen-atom abstraction, is now in close proximity to the nascent thioether sulfur and is ideally positioned for the second C-S bond forming event
malfunction
-
single mutations in the global regulator IscR can substantially improve cellular tolerance to BioB overexpression, increasing Escherichia coli dethiobiotin-to-biotin biocatalysis by more than 2.2fold. In the overexpression strains, FeS-cluster depletion is the main reason for toxicity. IscR mutations significantly affect cell viability and improve cell factories for de novo biosynthesis of thiamine by 1.3fold and lipoic acid by 1.8fold
physiological function
physiological function
-
biotin synthase catalyses the final step in the biosynthesis of biotin
physiological function
-
biotin synthase catalyses the last step in the biosynthesis of biotin
physiological function
-
biotin synthase is an AdoMet radical enzyme that catalyses the final step in the biosynthesis of biotin
physiological function
-
overexpression of biotin synthase is required for efficient generation of sulfur-35 labeled biotin in Escherichia coli. Coexpression of both BioB and the biotin ligase, BirA, is required for efficient biotinylation of biotin carboxy carrier protein-79
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
173900
-
homotetramer, determined by analytical ultracentrifugation, non-physiological form
38650 - 39670
-
wild-type and mutant enzymes, electrospray mass spectrometry
82000
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
additional information
-
2Fe-BioB, 188200, 14% tetramer, equilibrium analytical ultracentrifugation, tetrameric BioB is less active than dimeric enzyme, and it is assumed that the more abundant and active dimer reflects the native quaternary structure
dimer
-
2Fe-BioB, 92800, 86% dimer, equilibrium analytical ultracentrifugation
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C128A
C188A
C53A
C53S
-
no enzyme activity, spectrum shows no peak indicative of the presence of an [Fe-S] cluster
C57A
C57S
-
no enzyme activity, spectrum shows no peak indicative of the presence of an [Fe-S] cluster
C60A
C60S
-
no enzyme activity, spectrum shows no peak indicative of the presence of an [Fe-S] cluster
C97A
D155A
D155E
-
mutation in the highly conserved sequence motif, YNHNLD, in which Asn153 und Asp155 form hydrogen bonds with the ribose hydroxyl groups of AdoMet
D155N
-
mutation in the highly conserved sequence motif, YNHNLD, in which Asn153 und Asp155 form hydrogen bonds with the ribose hydroxyl groups of AdoMet
D155S
-
mutation in the highly conserved sequence motif, YNHNLD, in which Asn153 und Asp155 form hydrogen bonds with the ribose hydroxyl groups of AdoMet
H152A
N151A
N153A
N153D
-
mutation in the highly conserved sequence motif, YNHNLD, in which Asn153 und Asp155 form hydrogen bonds with the ribose hydroxyl groups of AdoMet
N153Q
-
mutation in the highly conserved sequence motif, YNHNLD, in which Asn153 und Asp155 form hydrogen bonds with the ribose hydroxyl groups of AdoMet
N153S
R260A
-
contains 1.8 iron atoms per monomer, slightly less active than wild-type
R260C
-
contains 2.0 iron atoms per monomer, slightly more active than wild-type
R260H
-
contains 1.3 iron atoms per monomer, slightly less active than wild-type
R260M
-
contains 1.1 iron atoms per monomer, about 30% of wild-type activity
C188A
-
cysteine desulfurase activity is reduced to 70% of the wild-type activity
C53A
-
inactive, but still exhibits UV-visible spectrum of a [Fe2-S2] cluster similar to that of the wild-type enzyme
C57A
-
inactive, but still exhibits UV-visible spectrum of a [Fe2-S2] cluster similar to that of the wild-type enzyme
C60A
-
inactive, but still exhibits UV-visible spectrum of a [Fe2-S2] cluster similar to that of the wild-type enzyme
D155A
-
no enzymic activity, mutant is unable to cleave S-adenosyl-l-methionine and to produce the deoxyadenosyl radical
D155A
-
mutation in the highly conserved sequence motif, YNHNLD, in which Asn153 und Asp155 form hydrogen bonds with the ribose hydroxyl groups of AdoMet
H152A
-
about 10% of turnover rate of wild-type. Ratio of 5-deoxyadenosine to biotin is about twice as high as in wild-type
N151A
-
no enzymic activity, mutant is unable to cleave S-adenosyl-l-methionine and to produce the deoxyadenosyl radical
N153A
-
no enzymic activity, mutant is unable to cleave S-adenosyl-l-methionine and to produce the deoxyadenosyl radical
N153A
-
mutation in the highly conserved sequence motif, YNHNLD, in which Asn153 und Asp155 form hydrogen bonds with the ribose hydroxyl groups of AdoMet
N153S
-
inactive for biotin production, enzyme can bind AdoMet and DTB and can convert DTB to 9-mercaptodethiobiotin, the majority of 9- mercaptodethiobiotin is released into the buffer and not bound to the enzyme, suggesting that dissociation of 9- mercaptodethiobiotin is faster than the second half reaction for biotin formation
N153S
-
mutation in the highly conserved sequence motif, YNHNLD, in which Asn153 und Asp155 form hydrogen bonds with the ribose hydroxyl groups of AdoMet
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
The loss of each FeS cluster results in a corresponding moderate decrease in the thermal stability, loss of the [4Fe-4S]2+ cluster and the bound substrates results in a 7°C decrease in stability
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
loss of the [2Fe-2S]2+ cluster from BioB does not result in global unfolding of the polypeptide chain at 20°C and does not result in significant destabilization of the dimer interface
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
-
-
His6-BioB is purified from Escherichia coli K12 WT strain BW25113, or from knockout strains lacking HscA, HscB, or DnaK, purification on a Ni-NTA-agarose column
-
wild-type and mutants N151A, H152A, N153A, D155A. Purified enzymes have a reddish brown colour
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
hexahistidine tagged proteins are expressed in Escherichia coli BL21DE3 pLysS cells
-
The delta pdxH::CAT allele of strain TX2767 is transduced into strain ER47 with phage P1vir to give strain AH14 into which plasmids pER30 and pER35 are introduced
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Shiuan, D.; Campbell, A.
Transcriptional regulation and gene arrangement of Escherichia coli, Citrobacter freundii and Salmonella typhimurium biotin operons
Gene
67
203-211
1988
Citrobacter freundii, Escherichia coli
Flint, D.H.; Allen, R.M.
Purification and characterization of biotin synthases
Methods Enzymol.
279
349-356
1997
Lysinibacillus sphaericus, Escherichia coli
Birch, O.M.; Fuhrmann, M.; Shaw, N.M.
Biotin synthase from Escherichia coli, an investigation of the low molecular weight and protein components required for activity in vitro
J. Biol. Chem.
270
19158-19165
1995
Escherichia coli
Sanyal, I.; Cohen, G.; Flint, D.H.
Biotin synthase: purification, characterization as a [2Fe-2S]cluster protein, and in vitro activity of the Escherichia coli bioB gene product
Biochemistry
33
3625-3631
1994
Escherichia coli
Bui, B.T.S.; Florentin, D.; Fournier, F.; Ploux, O.; Mejean, A.; Marquet, A.
Biotin synthase mechanism: on the origin of sulfur
FEBS Lett.
440
226-230
1998
Lysinibacillus sphaericus, Escherichia coli
Sanyal, I.; Gibson, K.J.; Flint, D.H.
Escherichia coli biotin synthase: an investigation into the factors required for its activity and its sulfur donor
Arch. Biochem. Biophys.
326
48-56
1996
Escherichia coli
Shaw, N.M.; Birch, O.M.; Tinschert, A.; Venetz, V.; Dietrich, R.; Savoy, L.A.
Biotin synthase from Escherichia coli: isolation of an enzyme-generated intermediate and stoichiometry of S-adenosylmethionine use
Biochem. J.
330
1079-1085
1998
Escherichia coli
Ugulava, N.B.; Sacanell, C.J.; Jarrett, J.T.
Spectroscopic changes during a single turnover of biotin synthase: Destruction of a [2Fe-2S] cluster accompanies sulfur insertion
Biochemistry
40
8352-8358
2001
Escherichia coli
Berkovitch, F.; Nicolet, Y.; Wan, J.T.; Jarrett, J.T.; Drennan, C.L.
Crystal structure of biotin synthase, an S-adenosylmethionine-dependent radical enzyme
Science
303
76-79
2004
Escherichia coli (P12996), Escherichia coli
Cosper, M.M.; Cosper, N.J.; Hong, W.; Shokes, J.E.; Broderick, W.E.; Broderick, J.B.; Johnson, M.K.; Scott, R.A.
Structural studies of the interaction of S-adenosylmethionine with the [4Fe-4S] clusters in biotin synthase and pyruvate formate-lyase activating enzyme
Protein Sci.
12
1573-1577
2003
Escherichia coli
Ugulava, N.B.; Surerus, K.K.; Jarrett, J.T.
Evidence from Mossbauer spectroscopy for distinct [2Fe-2S]2+ and [4Fe-4S]2+ cluster binding sites in biotin synthase from Escherichia coli
J. Am. Chem. Soc.
124
9050-9051
2002
Escherichia coli
Birch, O.M.; Hewitson, K.S.; Fuhrmann, M.; Burgdorf, K.; Baldwin, J.E.; Roach, P.L.; Shaw, N.M.
MioC is an FMN-binding protein that is essential for Escherichia coli biotin synthase activity in vitro
J. Biol. Chem.
275
32277-32280
2000
Escherichia coli
Bui, B.T.S.; Benda, R.; Schuenemann, V.; Florentin, D.; Trautwein, A.X.; Marquet, A.
Fate of the (2Fe-2S)2+ cluster of Escherichia coli biotin synthase during reaction: A Moessbauer characterization
Biochemistry
42
8791-8798
2003
Escherichia coli
Cosper, M.M.; Jameson, G.N.; Eidsness, M.K.; Huynh, B.H.; Johnson, M.K.
Recombinant Escherichia coli biotin synthase is a [2Fe-2S](2+) protein in whole cells
FEBS Lett.
529
332-336
2002
Escherichia coli
Cosper, M.M.; Jameson, G.N.L.; Davydov, R.; Eidsness, M.K.; Hoffman, B.M.; Huynh, B.H.; Johnson, M.K.
The [4Fe-4S]2+ cluster in reconstituted biotin synthase binds S-adenosyl-L-methionine
J. Am. Chem. Soc.
124
14006-14007
2002
Escherichia coli
Farh, L.; Hwang, S.Y.; Steinrauf, L.; Chiang, H.J.; Shiuan, D.
Structure-function studies of Escherichia coli biotin synthase via a chemical modification and site-directed mutagenesis approach
J. Biochem.
130
627-635
2001
Escherichia coli
Hewitson, K.S.; Baldwin, J.E.; Shaw, N.M.; Roach, P.L.
Mutagenesis of the proposed iron-sulfur cluster binding ligands in Escherichia coli biotin synthase
FEBS Lett.
466
372-376
2000
Escherichia coli
Kiyasu, T.; Asakura, A.; Nagahashi, Y.; Hoshino, T.
Biotin synthase of Bacillus subtilis shows less reactivity than that of Escherichia coli in in vitro reaction systems
Arch. Microbiol.
179
26-32
2002
Bacillus subtilis, Escherichia coli
McIver, L.; Baxter, R.L.; Campopiano, D.J.
Identification of the [Fe-S] cluster-binding residues of Escherichia coli biotin synthase
J. Biol. Chem.
275
13888-13894
2000
Escherichia coli
Ollagnier-de Choudens, S.; Sanakis, Y.; Hewitson, K.S.; Roach, P.; Munck, E.; Fontecave, M.
Reductive cleavage of S-adenosylmethionine by biotin synthase from Escherichia coli
J. Biol. Chem.
277
13449-13454
2002
Escherichia coli
Ollagnier-de-Choudens, S.; Mulliez, E.; Fontecave, M.
The PLP-dependent biotin synthase from Escherichia coli: mechanistic studies
FEBS Lett.
532
465-468
2002
Escherichia coli
Ollagnier-de-Choudens, S.; Mulliez, E.; Hewitson, K.S.; Fontecave, M.
Biotin synthase is a pyridoxal phosphate-dependent cysteine desulfurase
Biochemistry
41
9145-9152
2002
Escherichia coli
Ugulava, N.B.; Frederick, K.K.; Jarrett, J.T.
Control of adenosylmethionine-dependent radical generation in biotin synthase: A kinetic and thermodynamic analysis of substrate binding to active and inactive forms of BioB
Biochemistry
42
2708-2719
2003
Escherichia coli
Ugulava, N.B.; Gibney, B.R.; Jarrett, J.T.
Biotin synthase contains two distinct iron-sulfur cluster binding sites: Chemical and spectroelectrochemical analysis of iron-sulfur cluster interconversions
Biochemistry
40
8343-8351
2001
Escherichia coli
Kiyasu, T.; Asakura, A.; Nagahashi, Y.; Hoshino, T.
Contribution of cysteine desulfurase (NifS protein) to the biotin synthase reaction of Escherichia coli
J. Bacteriol.
182
2879-2885
2000
Escherichia coli
Mihara, H.; Esaki, N.
Bacterial cysteine desulfurases: their function and mechanisms
Appl. Microbiol. Biotechnol.
60
12-23
2002
Escherichia coli
Gibson, K.J.; Pelletier, D.A.; Turner, I.M., Sr.
Transfer of sulfur to biotin from biotin synthase (BioB protein)
Biochem. Biophys. Res. Commun.
254
632-635
1999
Escherichia coli
Tse Sum Bui, B.; Lotierzo, M.; Escalettes, F.; Florentin, D.; Marquet, A.
Further investigation on the turnover of Escherichia coli biotin synthase with dethiobiotin and 9-mercaptodethiobiotin as substrates
Biochemistry
43
16432-16441
2004
Escherichia coli
Mader Cosper, M.; Jameson, G.N.L.; Hernandez, H.L.; Krebs, C.; Huynh, B.H.; Johnson, M.K.
Characterization of the cofactor composition of Escherichia coli biotin synthase
Biochemistry
43
2007-2021
2004
Escherichia coli
Jameson, G.N.; Cosper, M.M.; Hernandez, H.L.; Johnson, M.K.; Huynh, B.H.
Role of the [2Fe-2S] cluster in recombinant Escherichia coli biotin synthase
Biochemistry
43
2022-2031
2004
Escherichia coli
Choi-Rhee, E.; Cronan, J.E.
Biotin synthase is catalytic in vivo, but catalysis engenders destruction of the protein
Chem. Biol.
12
461-468
2005
Escherichia coli
Tse Sum Bui, B.; Florentin, D.; Marquet, A.; Benda, R.; Trautwein, A.X.
Mossbauer studies of Escherichia coli biotin synthase: evidence for reversible interconversion between [2Fe-2S]2+ and [4Fe-4S]2+ clusters
FEBS Lett.
459
411-414
1999
Escherichia coli
Lotierzo, M.; Raux, E.; Tse Sum Bui, B.; Goasdoue, N.; Libot, F.; Florentin, D.; Warren, M.J.; Marquet, A.
Biotin synthase mechanism: mutagenesis of the YNHNLD conserved motif
Biochemistry
45
12274-12281
2006
Escherichia coli
Broach, R.B.; Jarrett, J.T.
Role of the [2Fe-2S]2+ cluster in biotin synthase: mutagenesis of the atypical metal ligand arginine 260
Biochemistry
45
14166-14174
2006
Escherichia coli
Tse Sum Bui, B.; Mattioli, T.A.; Florentin, D.; Bolbach, G.; Marquet, A.
Escherichia coli biotin synthase produces selenobiotin. Further evidence of the involvement of the [2Fe-2S]2+ cluster in the sulfur insertion step
Biochemistry
45
3824-3834
2006
Escherichia coli
Reyda, M.R.; Dippold, R.; Dotson, M.E.; Jarrett, J.T.
Loss of iron-sulfur clusters from biotin synthase as a result of catalysis promotes unfolding and degradation
Arch. Biochem. Biophys.
471
32-41
2008
Escherichia coli
Taylor, A.M.; Farrar, C.E.; Jarrett, J.T.
9-Mercaptodethiobiotin is formed as a competent catalytic intermediate by Escherichia coli biotin synthase
Biochemistry
47
9309-9317
2008
Escherichia coli
Abdel-Hamid, A.M.; Cronan, J.E.
In vivo resolution of conflicting in vitro results: Synthesis of biotin from dethiobiotin does not require pyridoxal phosphate
Chem. Biol.
14
1215-1220
2007
Escherichia coli
Lotierzo, M.; Bui, B.T.; Leech, H.K.; Warren, M.J.; Marquet, A.; Rigby, S.E.
Iron-sulfur cluster dynamics in biotin synthase: a new [2Fe-2S](1+) cluster
Biochem. Biophys. Res. Commun.
381
487-490
2009
Escherichia coli
Reyda, M.R.; Fugate, C.J.; Jarrett, J.T.
A complex between biotin synthase and the iron-sulfur cluster assembly chaperone HscA that enhances in vivo cluster assembly
Biochemistry
48
10782-10792
2009
Escherichia coli
Farrar, C.; Jarrett, J.
Protein residues that control the reaction trajectory in S-adenosylmethionine radical enzymes: Mutagenesis of asparagine 153 and aspartate 155 in Escherichia coli biotin synthase
Biochemistry
48
2448-2458
2009
Escherichia coli
Fuchs, M.G.; Meyer, F.; Ryde, U.
A combined computational and experimental investigation of the [2Fe-2S] cluster in biotin synthase
J. Biol. Inorg. Chem.
15
203-212
2010
Escherichia coli (P12996)
Delli-Bovi, T.; Spalding, M.; Prigge, S.
Overexpression of biotin synthase and biotin ligase is required for efficient generation of sulfur-35 labeled biotin in E. coli
BMC Biotechnol.
10
73
2010
Escherichia coli
Farrar, C.E.; Siu, K.K.; Howell, P.L.; Jarrett, J.T.
Biotin synthase exhibits burst kinetics and multiple turnovers in the absence of inhibition by products and product-related biomolecules
Biochemistry
49
9985-9996
2010
Escherichia coli
Fugate, C.J.; Jarrett, J.T.
Biotin synthase: insights into radical-mediated carbon-sulfur bond formation
Biochim. Biophys. Acta
1824
1213-1222
2012
Escherichia coli (P12996)
Fugate, C.J.; Stich, T.A.; Kim, E.G.; Myers, W.K.; Britt, R.D.; Jarrett, J.T.
9-Mercaptodethiobiotin is generated as a ligand to the [2Fe-2S]+ cluster during the reaction catalyzed by biotin synthase from Escherichia coli
J. Am. Chem. Soc.
134
9042-9045
2012
Escherichia coli
Kamachi, T.; Kouno, T.; Doitomi, K.; Yoshizawa, K.
Generation of adenosyl radical from S-adenosylmethionine (SAM) in biotin synthase
J. Inorg. Biochem.
105
850-857
2011
Escherichia coli (P12996)
Tao, L.; Stich, T.A.; Fugate, C.J.; Jarrett, J.T.; Britt, R.D.
EPR-derived structure of a paramagnetic intermediate generated by biotin synthase BioB
J. Am. Chem. Soc.
140
12947-12963
2018
Escherichia coli (P12996)
Bali, A.P.; Lennox-Hvenekilde, D.; Myling-Petersen, N.; Buerger, J.; Salomonsen, B.; Gronenberg, L.S.; Sommer, M.O.A.; Genee, H.J.
Improved biotin, thiamine, and lipoic acid biosynthesis by engineering the global regulator IscR
Metab. Eng.
60
97-109
2020
Escherichia coli, Escherichia coli BW25113
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