Information on EC 2.8.1.6 - biotin synthase

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The expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY
2.8.1.6
-
RECOMMENDED NAME
GeneOntology No.
biotin synthase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
dethiobiotin + sulfur-(sulfur carrier) + 2 S-adenosyl-L-methionine = biotin + (sulfur carrier) + 2 L-methionine + 2 5'-deoxyadenosine
show the reaction diagram
the reaction proceeds in two distinct steps involving mercaptodethiobiotin as an intermediate
Lavandula vera
-
dethiobiotin + sulfur-(sulfur carrier) + 2 S-adenosyl-L-methionine = biotin + (sulfur carrier) + 2 L-methionine + 2 5'-deoxyadenosine
show the reaction diagram
radical chemistry is involved in the mechanism of biotin synthase
-
dethiobiotin + sulfur-(sulfur carrier) + 2 S-adenosyl-L-methionine = biotin + (sulfur carrier) + 2 L-methionine + 2 5'-deoxyadenosine
show the reaction diagram
mechanism
-
dethiobiotin + sulfur-(sulfur carrier) + 2 S-adenosyl-L-methionine = biotin + (sulfur carrier) + 2 L-methionine + 2 5'-deoxyadenosine
show the reaction diagram
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 = biotin + (sulfur carrier) + 2 L-methionine + 2 5'-deoxyadenosine
show the reaction diagram
reaction proceeds via intermediate involving the (2Fe-2S) cluster, with a rate-determining step following the formation of this intermediate
-
dethiobiotin + sulfur-(sulfur carrier) + 2 S-adenosyl-L-methionine = biotin + (sulfur carrier) + 2 L-methionine + 2 5'-deoxyadenosine
show the reaction diagram
mechanism
-
dethiobiotin + sulfur-(sulfur carrier) + 2 S-adenosyl-L-methionine = biotin + (sulfur carrier) + 2 L-methionine + 2 5'-deoxyadenosine
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
sulfur atom transfer
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
biotin biosynthesis from 8-amino-7-oxononanoate I
-
biotin biosynthesis from 8-amino-7-oxononanoate II
-
Biotin metabolism
-
Metabolic pathways
-
SYSTEMATIC NAME
IUBMB Comments
dethiobiotin:sulfur sulfurtransferase
The enzyme binds a [4Fe-4S] and a [2Fe-2S] cluster. In every reaction cycle, the enzyme consumes two molecules of AdoMet, each producing 5'-deoxyadenosine and a putative dethiobiotinyl carbon radical. Reaction with another equivalent of AdoMet results in abstraction of the C6 methylene pro-S hydrogen atom from 9-mercaptodethiobiotin, and the resulting carbon radical is quenched via formation of an intramolecular C-S bond, thus closing the biotin thiophane 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.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
BIO2 protein (Arabidopsis thaliana clone pMP101 gene BIO2 reduced)
-
-
-
-
BioB
-
-
-
-
biotin synthase
-
-
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)
-
-
-
-
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
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
C41[DE3]pT7ec-bioB-1, which overexpresses Escherichia coli biotin synthase
-
-
Manually annotated by BRENDA team
host strain is blocked in the synthesis of pyridoxal (due to a null deletion/insertion mutation of pdxH which encodes pyridoxine 5-phosphate oxidase)
-
-
Manually annotated by BRENDA team
K12, strain 600
-
-
Manually annotated by BRENDA team
recombinant enzyme
-
-
Manually annotated by BRENDA team
Lavandula vera
-
-
-
Manually annotated by BRENDA team
recombinant strain BT(250)C
-
-
Manually annotated by BRENDA team
; strain W303-1A, W303-1B, BY4742
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
metabolism
-
biotin synthase catalyzes the final step in the biosynthesis of biotin
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
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
9-mercaptodethiobiotin + S-adenosyl-L-methionine
biotin + ?
show the reaction diagram
-
presence of S-adenosyl-L-methionine is required
-
-
?
9-mercaptodethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
9-mercaptodethiobiotin + [S] + S-adenosyl-L-methionine
biotin + ?
show the reaction diagram
-
-
-
-
?
9-mercaptodethiobiotin + [S] + S-adenosyl-L-methionine
biotin + L-methionine + 5'-deoxyadenosine
show the reaction diagram
-
-
-
-
ir
dethiobiotin + Na2S
biotin
show the reaction diagram
-
-
-
-
?
dethiobiotin + Na2Se
selenobiotin
show the reaction diagram
-
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 + S-adenosyl-L-methionine
biotin + ?
show the reaction diagram
-
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
-
-
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
P12996
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
O49543, Q8S904, Q8W3L1, Q9M0V0
-
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
insertion of a sulfur atom between the inactive methyl and methylene carbon atoms adjacent to the imidazolinone ring
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
ultimate sulfur donor has not yet been identified
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
Lavandula vera
-
intermediate is 9-mercaptodethiobiotin
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
insertion of a sulfur atom between the saturated C6 and C9 carbons of dethiobiotin
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
hypothesis: sulfur of biotin is derived from the [Fe-S] center of the enzyme
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
catalyzes the last step of the biosynthesis of biotin
-
-
-
dethiobiotin + sulfur
biotin
show the reaction diagram
-
catalyzes the last step of the biosynthesis of biotin
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
catalyzes the last step of the biosynthesis of biotin
-
-
-
dethiobiotin + sulfur
biotin
show the reaction diagram
-
catalyzes the last step of the biosynthesis of biotin
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
expression of the BIO2 gene appears to be induced under biotin-limiting conditions
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
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
show the reaction diagram
-
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
show the reaction diagram
-
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 + sulfur + S-adenosyl-L-methionine
biotin + L-methionine + 5'-deoxyadenosine
show the reaction diagram
-
-
-
-
?
dethiobiotin + [S] + 2 S-adenosyl-L-methionine
biotin + 2 L-methionine + 2 5'-deoxyadenosine
show the reaction diagram
-
-
-
-
ir
dethiobiotin + [S] + S-adenosyl-L-methionine
9-mercaptodethiobiotin + L-methionine + 5'-deoxyadenosine
show the reaction diagram
-
9-mercaptodethiobiotin is generated as a ligand to the [2Fe-2S]2+ cluster during the reaction
-
-
ir
dethiobiotin + [S] + S-adenosyl-L-methionine
biotin + L-methionine + 5'-deoxyadenosine
show the reaction diagram
-
-
-
-
?
dethiobiotin + [S] + S-adenosyl-L-methionine
biotin + L-methionine + 5'-deoxyadenosine
show the reaction diagram
-
-
-
-
ir
dethiobiotin + [S] + S-adenosyl-L-methionine
biotin + L-methionine + 5'-deoxyadenosine
show the reaction diagram
-
overall reaction
-
-
ir
dethiobiotin + [S] + S-adenosyl-L-methionine
biotin + L-methionine + 5'-deoxyadenosine
show the reaction diagram
-
dethiobiotin is 5-methyl-2-oxo-4-imidazolidinehexanoic acid
-
-
?
L-cysteine
L-alanine + sulfide
show the reaction diagram
-
enzyme displays cysteine desulfurase activity, providing it with the ability to mobilize sulfur from free cysteine
-
?
additional information
?
-
-
methionine and cysteine are no sulfur donors
-
-
-
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
?
-
Q9HF48
rate-limiting enzyme for biotin synthesis
-
-
-
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 SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
dethiobiotin + sulfur
biotin
show the reaction diagram
-
hypothesis: sulfur of biotin is derived from the [Fe-S] center of the enzyme
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
catalyzes the last step of the biosynthesis of biotin
-
-
-
dethiobiotin + sulfur
biotin
show the reaction diagram
-
catalyzes the last step of the biosynthesis of biotin
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
catalyzes the last step of the biosynthesis of biotin
-
-
-
dethiobiotin + sulfur
biotin
show the reaction diagram
-
catalyzes the last step of the biosynthesis of biotin
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
expression of the BIO2 gene appears to be induced under biotin-limiting conditions
-
-
?
dethiobiotin + sulfur
biotin
show the reaction diagram
-
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
show the reaction diagram
-
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
show the reaction diagram
-
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 + sulfur + S-adenosyl-L-methionine
biotin + L-methionine + 5'-deoxyadenosine
show the reaction diagram
-
-
-
-
?
additional information
?
-
Q9HF48
rate-limiting enzyme for biotin synthesis
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
FAD
-
enhances activity
Fe-S center
-
the enzyme contains one [4Fe-4S]2+ cluster and one air-stable [2Fe-2S]2+ cluster per monomer
Fe-S center
-
the enzyme contains a [2Fe-2S]2+ cluster
Fe-S center
-
the active site of the enzyme involves a [4Fe-4S] cluster
Ferredoxin
-
required
-
Flavodoxin
-
required
-
NADH
-
enhances activity
NADH
O49543, Q8S904, Q8W3L1, Q9M0V0
-
NADPH
-
enhances activity
NADPH
-
enhances activity
NADPH
-
omission results in a 100fold decrease in activity
NADPH
-
required
NADPH
O49543, Q8S904, Q8W3L1, Q9M0V0
-
S-adenosyl-L-methionine
-
-
thiamine diphosphate
-
required
additional information
-
not: pyridoxal 5-phosphate
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
enhances activity
Fe2+
-
required
Fe2+
-
enhances activity
Fe2+
-
enhances activity
Fe2+
-
highly stimulates
FeCl3
-
enhances activity 3-4fold at 0.1 mM
FeCl3
-
required for formation of mixed [Fe-S] cluster state
FeCl3
-
sulfur of the iron-sulfur cluster is provided by cysteine desulfurase EC 2.8.1.7
FeCl3
-
binds 1 [Fe4-S4] cluster per monomer; sulfur of the iron-sulfur cluster is provided by cysteine desulfurase EC 2.8.1.7
Iron
-
presence of a [2Fe-2S] cluster; the 104000 Da dimeric enzyme form contains a single [2Fe-2S] cluster per dimer; the 82000 Da dimeric enzyme form contains one [2Fe-2S]cluster per monomer
Iron
-
the 104000 Da dimeric enzyme form contains a single [2Fe-2S] cluster per dimer; the 82000 Da dimeric enzyme form contains one [2Fe-2S]cluster per monomer
Iron
-
the enzyme contains an iron-sulfur cluster
Iron
-
presence of a [2Fe-2S] cluster
Iron
-
1 [2Fe-2S] and 1 [4Fe-4S] cluster per monomer are essential for optimal activity
Iron
-
[2Fe-2S] and [4Fe-4S] cluster
Iron
-
[4Fe-4S] cluster
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; presence of a [2Fe-2S] cluster
Iron
-
[Fe4-S4] cluster binds S-adenosylmethionine
Iron
-
presence of a [2Fe-2S] cluster
Iron
-
the enzyme contains an iron-sulfur cluster
Iron
-
[4Fe-4S] is involved in cleavage of S-adenosylmethionine
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
-
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 dimmer containing 2 [Fe2-S2] clusters and 2 [Fe4-S4] clusters
Iron
-
the enzyme contains an iron-sulfur 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
-
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
-
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
-
the [2Fe-2S]2+ cluster is implicated in the sulfur insertion step
Iron
-
Fe/S cluster assembly of biotin synthase strongly depends on Isu1 and Isu2 proteins; Fe/S cluster assembly on Bio2 strongly depends on the Isu1 and Isu2 proteins, Isa proteins are crucial for the in vivo function of biotin synthase but not for the de novo synthesis of its Fe/S clusters.
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.; 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; the [2Fe-2S]2+ cluster is both the sulfur-donating substrate and the sulfur-oxidizing cofactor
Mn2+
-
enhances activity
Na2S
-
enhances activity 3-4fold at 1 mM
Na2S
-
required for formation of mixed [Fe-S] cluster state
S2-
-
enhances activity
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
5'-Deoxyadenosine
-
strong inhibitor, reversible, 90% inhibition upon addition of 1 equivalent of 5'-deoxyadenosine with regard to the enzyme
acidomycin
-
structural analog of biotin, complete inhibition at 0.4 mM, 50% inhibition at 0.035 mM
Cd2+
-
1 mM, almost complete inhibition
Co2+
-
1 mM, almost complete inhibition
Cu2+
-
1 mM, almost complete inhibition
Fe2+
-
above 1 mM
-
Hg2+
-
1 mM, almost complete inhibition
NaBH4
-
decreases cysteine desulfurase and biotin synthase activity
S-adenosyl-L-homocysteine
-
potent inhibitor
Zn2+
-
1 mM, almost complete inhibition
methionine and 5'-deoxyadenosine
-
modest inhibitor
-
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 ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Amino acid
-
one of the amino acids: Asn, Asp, Gln or Ser
asparagine
-
required
D-fructose 1,6-bisphosphate
-
enhances biotin formation
dithiothreitol
-
potential sulfur donor
dithiothreitol
-
-
dithiothreitol
-
10fold stimulation of cysteine desulfurase activity of the enzyme at 20 mM
dithiothreitol
-
required
dithiothreitol
-
required for formation of mixed [Fe-S] cluster state
Flavodoxin
-
-
-
flavodoxin reductase
-
omission results in a 100fold decrease in activity
-
flavodoxin reductase
-
-
-
fructose 1,6-diphosphate
-
-
L-cysteine
-
required as sulfur donor
L-cysteine
-
required
L-cysteine
-
increases biotin production
L-cysteine
-
highly stimulates
NifS
-
a member of the Nif protein family stimulates biotin production
-
NifU
-
a member of the Nif protein family stimulates biotin production
-
Pyridine nucleotide
-
required, NADPH being most effective
-
pyridoxal 5'-phosphate
-
required for cysteine desulfurase activity of the enzyme
S-adenosyl-L-methionine
-
absolute requirement as electron source
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
-
required
S-adenosyl-L-methionine
-
required
S-adenosyl-L-methionine
-
one molecule S-adenosyl-L-methionine is required to form one molecule of biotin
S-adenosyl-L-methionine
-
required
S-adenosyl-L-methionine
-
-
Thiamine diphosphate-dependent protein
-
required
-
MioC
-
essential, may function as an electron transport protein
-
additional information
-
a labile low-molecular-weight product of the 7,8-diaminoperlargonic acid aminotransferase reaction stimulates by 3fold
-
additional information
-
one or more unidentified factors from mitochoncrial matrix of pea and potato and from mitochondrial membranes of pea, in addition to the purified enzyme, are obligatory for the conversion of dithiobiotin to biotin in plants
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.005
-
9-Mercaptodethiobiotin
-
-
0.2
-
9-Mercaptodethiobiotin
-
pH and temperature not specified in the publication
0.0016
-
dethiobiotin
-
-
0.002
-
dethiobiotin
-
pH 7.5
0.002
-
dethiobiotin
-
-
0.005
-
dethiobiotin
-
pH and temperature not specified in the publication
0.03
-
dethiobiotin
-
pH 8, 37C
0.03
-
L-cysteine
-
pH 8, 37C
1
-
NADH
O49543, Q8S904, Q8W3L1, Q9M0V0
pH 8
0.0024
-
NADPH
O49543, Q8S904, Q8W3L1, Q9M0V0
pH 8
0.01
-
S-adenosyl-L-methionine
-
in 50 mM Tris-HCl, 100 mM KCl, and 5 mM dithiothreitol, pH 8.0, at 37C
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0000005
-
dethiobiotin
-
pH 7.5, 30C, cell free extract
0.0000142
-
dethiobiotin
-
pH 7.5, 30C, cell free extract
0.00055
-
dethiobiotin
-
pH 8, 37C
13.9
-
dethiobiotin
-
pH 7.5, 37C
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.02
-
methionine and 5'-deoxyadenosine
-
in 50 mM Tris-HCl, 100 mM KCl, and 5 mM dithiothreitol, pH 8.0, at 37C
-
0.00065
-
S-adenosyl-L-homocysteine
-
in 50 mM Tris-HCl, 100 mM KCl, and 5 mM dithiothreitol, pH 8.0, at 37C
0.075
-
Sinefungin
-
in 50 mM Tris-HCl, 100 mM KCl, and 5 mM dithiothreitol, pH 8.0, at 37C
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
-
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
8
-
-
activity assay
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7
9
-
pH 7: about 50% of maximal activity, pH 9: about 60% of maximal activity
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
activity assay
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
35
40
-
35C: about 70% of maximal activity, 40C: about 60% of maximal activity
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
Lavandula vera
-
-
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
O49543, Q8S904, Q8W3L1, Q9M0V0
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Escherichia coli (strain K12)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
22000
-
O49543, Q8S904, Q8W3L1, Q9M0V0
Adx1 isozyme, SDS-PAGE
38650
39670
-
wild-type and mutant enzymes, electrospray mass spectrometry
41650
-
-
calculated from nucleotide sequence
45000
-
-
His6-BioB, determined by SDS-PAGE and Western blot analysis
50000
-
O49543, Q8S904, Q8W3L1, Q9M0V0
AdxR isozyme, SDS-PAGE
75000
-
-
gel filtration
76000
-
-
dimeric form
80000
-
-
gel filtration
82000
-
-
at least 3 forms of the enzyme: 82000 Da, 104000 Da, 160000 Da
82000
-
-
gel electrophoresis
82630
-
-
homodimer, theoretical
90680
-
-
homodimer, determined by analytical ultracentrifugation
104000
-
-
at least 3 forms of the enzyme: 82000 Da, 104000 Da, 160000 Da
160000
-
-
at least 3 forms of the enzyme: 82000 Da, 104000 Da, 160000 Da
165300
-
-
homotetramer, theoretical
173900
-
-
homotetramer, determined by analytical ultracentrifugation, non-physiological form
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 38000, SDS-PAGE
dimer
-
2 * 36954, calculation from nucleotide sequence; 2 * 37000, SDS-PAGE
dimer
-
2 * 39000, 82000 MW form and 104000 MW form, SDS-PAGE
dimer
-
2 * 38700, calculation from nucleotide sequence; 2 * 39000, 82000 MW form and 104000 MW form, SDS-PAGE
dimer
-
2 * 38400
dimer
-
enzyme exists in 2 forms: a dimer and a monomer
dimer
-
2 * 37000, SDS-PAGE
dimer
-
homodimer
dimer
-
2Fe-BioB, 92800, 86% dimer, equilibrium analytical ultracentrifugation
homodimer
-
-
homodimer
-
2 * 38600, calculated from amino acid sequence
monomer
-
enzyme exists in 2 forms: a dimer and a monomer
additional information
-
Fe/S cluster assembly of biotin synthase strongly depends on Isu1 and Isu2 proteins. Proteins Isa1 and Isa2 are crucial foin vivo function of biotin synthase but not for de novo synthesis of its Fe/S clusters
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
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
complexed with S-adenosyl-L-methionine and dethiobiotin; in complex with S-adensyl-L-methionine and dethiobiotin
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
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 7C decrease in stability
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
loss of the [2Fe-2S]2+ cluster from BioB does not result in global unfolding of the polypeptide chain at 20C and does not result in significant destabilization of the dimer interface
-
loses all its clusters if the purification is carried out with buffers that have not been saturated with argon
-
OXIDATION STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
oxygen sensitive
-
645582
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-
O49543, Q8S904, Q8W3L1, Q9M0V0
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 mutant enzymes
-
wild-type and mutants N151A, H152A, N153A, D155A. Purified enzymes have a reddish brown colour
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expression in Escherichia coli. The amino-terminal portion of the BIO2 gene may play a role in localizing the BIO2 protein to a subcellular compartment
-
the primary sequence of the Arabidopsis biotin synthase is similar to biotin synthase from Escherichia coli, Serratia marcescens and Saccharomyces cerevisiae, about 50% sequence identity, and more distantly related to the Bacillus sphaericus enzyme, 33% sequence identity
-
expressed in Escherichia coli B834(DE3) cells
-
expressed in Escherichia coli BL21(DE3)pLysS cells
-
hexahistidine tagged protein is expressed in Escherichia coli cells
-
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
-
wild-type and mutants
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C128A
-
reduced enzyme activity
C128A
-
no enzyme activity
C128A
-
reduced cysteine desulfurase activity
C188A
-
reduced enzyme activity
C188A
-
no enzyme activity
C188A
-
cysteine desulfurase activity is reduced to 70% of the wild-type activity
C188S
-
enzyme is insoluble
C276A
-
normal enzyme activity
C288A
-
normal enzyme activity
C288T
-
normal enzyme activity
C53A
-
inactive, but still exhibits UV-visible spectrum of a [Fe2-S2] cluster similar to that of the wild-type enzyme
C53A
-
no enzyme activity
C53S
-
no enzyme activity, spectrum shows no peak indicative of the presence of an [Fe-S] cluster
C57A
-
inactive, but still exhibits UV-visible spectrum of a [Fe2-S2] cluster similar to that of the wild-type enzyme
C57A
-
no enzyme activity
C57S
-
no enzyme activity, spectrum shows no peak indicative of the presence of an [Fe-S] cluster
C60A
-
inactive, but still exhibits UV-visible spectrum of a [Fe2-S2] cluster similar to that of the wild-type enzyme
C60A
-
no enzyme activity
C60S
-
no enzyme activity, spectrum shows no peak indicative of the presence of an [Fe-S] cluster
C97A
-
no enzyme activity
C97A
-
reduced cysteine desulfurase activity
D155A
-
no enzymic activity, mutant is unable to cleave S-adenosyl-l-methionine and to produce the deoxyadenosyl radical
D155A
-
mutation in the conserved YNHNLD sequence
D155A
-
mutation in the highly conserved sequence motif, YNHNLD, in which Asn153 und Asp155 form hydrogen bonds with the ribose hydroxyl groups of AdoMet
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
-
about 10% of turnover rate of wild-type. Ratio of 5-deoxyadenosine to biotin is about twice as high as in wild-type
H152A
-
mutation in the conserved YNHNLD sequence
N151A
-
no enzymic activity, mutant is unable to cleave S-adenosyl-l-methionine and to produce the deoxyadenosyl radical
N151A
-
mutation in the conserved YNHNLD sequence
N153A
-
no enzymic activity, mutant is unable to cleave S-adenosyl-l-methionine and to produce the deoxyadenosyl radical
N153A
-
mutation in the conserved YNHNLD sequence
N153A
-
mutation in the highly conserved sequence motif, YNHNLD, in which Asn153 und Asp155 form hydrogen bonds with the ribose hydroxyl groups of AdoMet
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
-
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
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
additional information
Q9HF48
integration of BIO2 enzyme gene into chromosome under strong promoter. Biotin yield of system reaches 100fold above wild-type. Biotin production is not stable if the selection pressure is removed
R260M
-
contains 1.1 iron atoms per monomer, about 30% of wild-type activity
additional information
-
Fe/S cluster assembly of biotin synthase strongly depends on Isu1 and Isu2 proteins. Proteins Isa1 and Isa2 are crucial for in vivo function of biotin synthase but not for de novo synthesis of its Fe/S clusters
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
nutrition
Q9HF48
production of biotin-rich feed and food additives using integration of BIO2 enzyme gene into chromosome under strong promoter for increased production of biotin