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Information on EC 3.5.2.2 - dihydropyrimidinase and Organism(s) Geobacillus stearothermophilus and UniProt Accession Q45515
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3.5.2.2 dihydropyrimidinaseIUBMB Comments
Also acts on dihydrothymine and hydantoin.
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- 3.5.2.2
- 5-fluorouracil
- thymidylate
-
dihydropyridine
- colorectal
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fluoropyrimidine
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l-type
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pharmacogenetic
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1,4-dihydropyridine
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tegafur
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prodrugs
-
voltage-dependent
- dihydrouracil
-
5-fu-based
-
fluoropyrimidine-based
-
nitrendipine
- leucovorin
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phosphoribosyltransferase
- diltiazem
- oxaliplatin
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hemoperfusion
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phosphoribosyl
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5-fluorouracil-based
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pyrrolizidine
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excitation-contraction
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5-fu-induced
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dihydropyridine-sensitive
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5\'-dfur
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pretherapeutic
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omega-conotoxin
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hand-foot
The taxonomic range for the selected organisms is: Geobacillus stearothermophilus
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Synonyms
dhp, dihydropyrimidine dehydrogenase, dihydropyrimidinase, hydantoinase, d-hydantoinase, dhp-1, dhpase, dihydropyrimidine amidohydrolase, dhp-2, padhpase, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
D-hydantoinase
DHP
-
-
-
-
Dihydropyrimidinase
-
-
-
-
hydantoin peptidase
-
-
-
-
hydantoinase
hydropyrimidine hydrase
-
-
-
-
pyrimidine hydrase
-
-
-
-
D-hydantoinase
-
-
-
-
hydantoinase
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
carboxylic acid amide hydrolysis
-
-
-
-
PATHWAY SOURCE
PATHWAYS
KEGG
-
-, -
SYSTEMATIC NAME
IUBMB Comments
5,6-dihydropyrimidine amidohydrolase
Also acts on dihydrothymine and hydantoin.
CAS REGISTRY NUMBER
COMMENTARY
9030-74-4
-
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
DL-p-hydroxyphenylhydantoin + H2O
N-carbamoyl-D-hydroxyphenylglycine
-
-
-
?
5,6-dihydrouracil + H2O
3-ureidopropionate
-
-
-
-
?
D-5-(1-methylpropyl)hydantoin + H2O
N-carbamoyl-D-isoleucine
-
-
-
?
D-5-benzylhydantoin + H2O
N-carbamoyl-D-phenylalanine
-
-
-
?
D-5-isobutylhydantoin + H2O
N-carbamoyl-D-leucine
-
-
-
?
D-5-isopropylhydantoin + H2O
N-carbamoyl-D-valine
-
-
-
?
D-5-methylhydantoin + H2O
N-carbamoyl-D-alanine
-
-
-
?
DL-5-(p-hydroxyphenyl)hydantoin + H2O
N-carbamoyl-p-hydroxyphenylglycine
-
-
-
-
?
DL-5-phenylhydantoin + H2O
N-carbamoyl-D-phenylglycine
-
-
-
-
?
DL-hydroxyphenylhydantoin + H2O
N-carbamoyl-D-hydroxyphenylglycine
-
-
-
-
?
DL-phenylhydantoin + H2O
N-carbamoyl-D-phenylglycine
-
-
-
-
?
hydantoin + H2O
hydantoic acid
-
-
-
-
?
hydantoin + H2O
N-carbamoyl glycine
-
-
-
-
?
L-5-(1-methylpropyl)hydantoin + H2O
N-carbamoyl-L-isoleucine
-
-
-
?
L-5-benzylhydantoin + H2O
N-carbamoyl-L-phenylalanine
-
-
-
?
L-5-isobutylhydantoin + H2O
N-carbamoyl-L-leucine
-
-
-
?
L-5-methylhydantoin + H2O
N-carbamoyl-L-alanine
-
-
-
?
D-5-(2-methylthioethyl)hydantoin + H2O
N-carbamoyl-D-methionine
-
-
-
?
D-5-(2-methylthioethyl)hydantoin + H2O
N-carbamoyl-D-methionine
-
-
-
-
?
L-5-(2-methylthioethyl)hydantoin + H2O
N-carbamoyl-L-methionine
-
-
-
?
L-5-(2-methylthioethyl)hydantoin + H2O
N-carbamoyl-L-methionine
-
-
-
?
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
5,6-dihydrouracil + H2O
3-ureidopropionate
-
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Co2+
Mn2+
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2.9
DL-p-hydroxyphenylhydantoin
mutant M63I/F159S, pH 8.0, 37°C
4.9
DL-p-hydroxyphenylhydantoin
wild-type, pH 8.0, 37°C
5.1
DL-p-hydroxyphenylhydantoin
mutant M63H/F159N, pH 8.0, 37°C
2.2
DL-hydroxyphenylhydantoin
-
pH 8.0, 37°C, mutant enzyme F159L
3.5
DL-hydroxyphenylhydantoin
-
pH 8.0, 37°C, mutant enzyme F159I
3.6
DL-hydroxyphenylhydantoin
-
pH 8.0, 37°C, mutant enzyme F159V
5.4
DL-hydroxyphenylhydantoin
-
pH 8.0, 37°C, wild-type enzyme
7.4
DL-hydroxyphenylhydantoin
-
pH 8.0, 37°C, mutant enzyme F159A
13
DL-hydroxyphenylhydantoin
-
pH 8.0, 37°C, mutant enzyme L65F/F159A
24
DL-hydroxyphenylhydantoin
-
pH 8.0, 37°C, mutant enzyme M63F/L65V
27
DL-hydroxyphenylhydantoin
-
pH 8.0, 37°C, mutant enzyme F159S
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
59
DL-p-hydroxyphenylhydantoin
mutant M63I/F159S, pH 8.0, 37°C
62
DL-p-hydroxyphenylhydantoin
mutant M63H/F159N, pH 8.0, 37°C
11
DL-hydroxyphenylhydantoin
-
pH 8.0, 37°C, mutant enzyme F159L
46
DL-hydroxyphenylhydantoin
-
pH 8.0, 37°C, mutant enzyme F159I
50
DL-hydroxyphenylhydantoin
-
pH 8.0, 37°C, mutant enzyme F159V
110
DL-hydroxyphenylhydantoin
-
pH 8.0, 37°C, mutant enzyme F159A
180
DL-hydroxyphenylhydantoin
-
pH 8.0, 37°C, mutant enzyme M63F/L65V
200
DL-hydroxyphenylhydantoin
-
pH 8.0, 37°C, mutant enzyme L65F/F159A
280
DL-hydroxyphenylhydantoin
-
pH 8.0, 37°C, mutant enzyme F159S
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
3.63
DL-p-hydroxyphenylhydantoin
wild-type, pH 8.0, 37°C
12.17
DL-p-hydroxyphenylhydantoin
mutant M63H/F159N, pH 8.0, 37°C
20.95
DL-p-hydroxyphenylhydantoin
mutant M63I/F159S, pH 8.0, 37°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
9.5
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
65
70
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
200000
60000
-
2 * 60000 + 2 * 62000 + 1 * 110000, room temperature, 2-mercaptoethanol, SDS-PAGE
62000
-
2 * 60000 + 2 * 62000 + 1 * 110000, room temperature, 2-mercaptoethanol, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
dimer
oligomer
-
2 * 60000 + 2 * 62000 + 1 * 110000, room temperature, 2-mercaptoethanol, SDS-PAGE
tetramer
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
apocrystal structure is determined at 3.0 A resolution, crystals grow at 21°C from hanging drops by the vapor diffusion method
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
F159A
-
turnover number for hydantoin is 10% of that of the wild-type enzyme, Km-value for hydantoin is 4.6fold higher than that of the wild-type enzyme, turnover-number for DL-hydroxyphenylhydantoin is 6.1fold higher than that of the wild-type enzyme, Km-value for DL-hydroxyphenylhydantoin is 1.4fold higher than that of the wild-type enzyme. Specificity of the mutant enzyme towards aromatic substrate hydroxyphenylhydantoin is enhanced by 200fold compared with that of the wild-type enzyme
F159I
-
turnover number for hydantoin is 61% of that of the wild-type enzyme, Km-value for hydantoin is 1.9fold higher than that of the wild-type enzyme, turnover-number for DL-hydroxyphenylhydantoin is 2.6fold higher than that of the wild-type enzyme, Km-value for DL-hydroxyphenylhydantoin is 1.5fold lower than that of the wild-type enzyme
F159L
-
turnover number for hydantoin 64% of the turnover number of the wild-type enzyme, Km-value for hydantoin is 90% of the value of the wild-type enzyme, turnover-number for DL-hydroxyphenylhydantoin is 61% of that of the wild-type enzyme, Km-value for DL-hydroxyphenylhydantoin is 2.5fold lower than that of the wild-type enzyme
F159S
-
turnover number for hydantoin is 4.7% of that of the wild-type enzyme, Km-value for hydantoin is 4fold higher than that of the wild-type enzyme, turnover-number for DL-hydroxyphenylhydantoin is 15.5fold higher than that of the wild-type enzyme, Km-value for DL-hydroxyphenylhydantoin is 5fold higher than that of the wild-type enzyme
F159V
-
turnover number for hydantoin is 22% of that of the wild-type enzyme, Km-value for hydantoin is 1.6fold higher than that of the wild-type enzyme, turnover-number for DL-hydroxyphenylhydantoin is 2.8fold higher than that of the wild-type enzyme, Km-value for DL-hydroxyphenylhydantoin is 1.5fold lower than that of the wild-type enzyme
L65F/F159A F159A
-
turnover number for hydantoin is 10.9% of that of the wild-type enzyme, Km-value for hydantoin is 4.8fold higher than that of the wild-type enzyme, turnover-number for DL-hydroxyphenylhydantoin is 11fold higher than that of the wild-type enzyme, Km-value for DL-hydroxyphenylhydantoin is 2.4fold higher than that of the wild-type enzyme. Specificity of the mutant enzyme towards aromatic substrate hydroxyphenylhydantoin is enhanced by 200fold compared with that of the wild-type enzyme
M63F/L65V F159A
-
turnover number for hydantoin is 9.5% of that of the wild-type enzyme, Km-value for hydantoin is 2.6fold higher than that of the wild-type enzyme, turnover-number for DL-hydroxyphenylhydantoin is 10fold higher than that of the wild-type enzyme, Km-value for DL-hydroxyphenylhydantoin is 4.4fold higher than that of the wild-type enzyme. Specificity of the mutant enzyme towards aromatic substrate hydroxyphenylhydantoin is enhanced by 200fold compared with that of the wild-type enzyme
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
40 - 80
-
fairly thermostable, oligomer retains full activity at 40-60°C, still retains 98% initial activity at 70°C
60
80
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene for thermostable hydantoinase cloned and overexpressed in Escherichia coli JM105, vector pAHN134
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
synthesis
to produce optically pure D-amino acids, microbial D-hydantoinase is used for stereospecific hydrolysis of chemically synthesized cyclic hydantoins
synthesis
synthesis
-
artificial fusion enzyme useful as potential biocatalyst for production of nonnatural amino acids
synthesis
-
commercial production of optically pure amino acids for synthesis of antibiotics, pharmaceuticals, artificial sweeteners and biologically active peptides, food ingredients, pyrethroids, pesticides, antimicrobial and antiviral agents and other agrochemicals
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Ishikawa, T.; Mukohara, Y.; Watabe, K.; Kobayashi, S.; Nakamura, H.
Microbial conversion of DL-5-substituted hydantoins to the corresponding L-amino acids by Bacillus stearothermophilus NS1122A
Biosci. Biotechnol. Biochem.
58
265-270
1994
Arthrobacter sp., Geobacillus stearothermophilus, Bos taurus, Flavobacterium sp., Pseudomonas sp., Pseudomonas putida, Pseudomonas fluorescens, Thermus sp., Pseudomonas sp. NS671, Arthrobacter sp. DSM 3747, Flavobacterium sp. AJ-3912
-
Mukohara, Y.; Ishikawa, T.; Watabe, K.; Nakamura, H.
A thermostable hydantoinase of Bacillus stearothermophilus NS1122A: Cloning, sequencing, and high expression of the enzyme gene, and some properties of the expressed enzyme
Biosci. Biotechnol. Biochem.
58
1621-1626
1994
Geobacillus stearothermophilus, Bos taurus, Pseudomonas sp., Pseudomonas putida, Pseudomonas sp. NS671
Luksa, V.; Starkuviene, V.; Starkuviene, B.; Dagys, R.
Purification and characterization of the D-hydantoinase from Bacillus circulans
Appl. Biochem. Biotechnol.
62
219-230
1997
Agrobacterium sp., Arthrobacter crystallopoietes, Geobacillus stearothermophilus, Brevibacillus brevis, Niallia circulans, Pseudomonas sp., Pseudomonas putida, Pseudomonas fluorescens, Agrobacterium sp. IP-I 671
Seung-Goo, L.; Dong-Cheol, L.; Hak-Sung, K.
Purification and characterization of thermostable D-hydantoinase from thermophilic Bacillus stearothermophilus SD-1
Appl. Biochem. Biotechnol.
62
251-266
1997
Agrobacterium sp., Geobacillus stearothermophilus, Bos taurus, Pseudomonas putida, Pseudomonas fluorescens, Rattus norvegicus, Thermus sp., Geobacillus stearothermophilus SD-1
May, O.; Siemann, M.; Pietzsch, M.; Kiess, M.; Mattes, R.; Syldatk, C.
Substrate-dependent enantioselectivity of a novel hydantoinase from Arthrobacter aurescens DSM 3745: Purification and characterization as new member of cyclic amidases
J. Biotechnol.
61
1-13
1998
Agrobacterium tumefaciens, Paenarthrobacter aurescens, Arthrobacter sp., Geobacillus stearothermophilus, Brevibacillus brevis, Bos taurus, Flavobacterium sp., Pseudomonas putida, Pseudomonas stutzeri, Rattus norvegicus, Geobacillus stearothermophilus JC2310, Arthrobacter sp. DSM 3747
Joo-Ho, P.; Geun-Joong, K.; Seun-Goo, L.; Dong-Cheol, L.; Hak-Sung, K.
Purification and characterization of thermostable D-hydantoinase from Bacillus thermocatenulatus GH-2
Appl. Biochem. Biotechnol.
81
53-64
1999
Agrobacterium sp., Geobacillus stearothermophilus, Geobacillus thermocatenulatus, Pseudomonas sp., Geobacillus stearothermophilus SD-1, Geobacillus thermocatenulatus GH-2
Sharma, R.; Vohra, R.M.
A thermostable D-hydantoinase isolated from a mesophilic Bacillus sp. AR9
Biochem. Biophys. Res. Commun.
234
485-488
1997
Agrobacterium tumefaciens, Agrobacterium sp., Geobacillus stearothermophilus, Bacillus sp. (in: Bacteria), Bos taurus, Peptococcus anaerobius, Pseudomonas sp., Pseudomonas putida, Geobacillus stearothermophilus SD-1, Bacillus sp. (in: Bacteria) AR9
Geun-Joong, K.; Hak-Sung, K.
C-terminal regions of D-hydantoinases are nonessential for catalysis, but affect the oligomeric structure
Biochem. Biophys. Res. Commun.
243
96-100
1998
Geobacillus stearothermophilus, Geobacillus thermocatenulatus, Homo sapiens, Pseudomonas putida, Rattus norvegicus, Geobacillus stearothermophilus SD-1, Geobacillus thermocatenulatus GH-2
Siemann, M.; Alvarado-Marin, A.; Pietzsch, M.; Syldatk, C.
A D-specific hydantoin amidohydrolase: properties of the metalloenzyme purified from Arthrobacter crystallopoietes
J. Mol. Catal. , B Enzym.
6
387-397
1999
Agrobacterium tumefaciens, Agrobacterium sp., Paenarthrobacter aurescens, Arthrobacter crystallopoietes, Geobacillus stearothermophilus, Bacillus sp. (in: Bacteria), Parageobacillus thermoglucosidasius, Homo sapiens, Pseudomonas sp., Pseudomonas putida, Pseudomonas fluorescens, Rattus norvegicus, Agrobacterium sp. IP-I 671, Pseudomonas sp. AJ-11220, Bacillus sp. (in: Bacteria) LU 1220
-
Geun-Joong, K.; Dong-Eun, L.; Hak-Sung, K.
Construction and evaluation of a novel bifunctional N-carbamylase-D-hydantoinase fusion enzyme
Appl. Environ. Microbiol.
66
2133-2138
2000
Agrobacterium tumefaciens, Agrobacterium sp., Geobacillus stearothermophilus, Geobacillus thermocatenulatus, Escherichia coli, Homo sapiens, Pseudomonas putida, Agrobacterium sp. IP-I 671, Agrobacterium tumefaciens DSM 30147, Geobacillus stearothermophilus SD-1, Geobacillus thermocatenulatus GH-2
Cheon, Y.H.; Park, H.S.; Kim, J.H.; Kim, Y.; Kim, H.S.
Manipulation of the active site loops of D-hydantoinase, a (beta/alpha)8-barrel protein, for modulation of the substrate specificity
Biochemistry
43
7413-7420
2004
Geobacillus stearothermophilus, Geobacillus stearothermophilus SD1
Lin, L.L.; Hsu, W.H.; Hsu, W.Y.; Kan, S.C.; Hu, H.Y.
Phylogenetic analysis and biochemical characterization of a thermostable dihydropyrimidinase from alkaliphilic Bacillus sp. TS-23
Antonie van Leeuwenhoek
88
189-197
2005
Thermus sp., Streptomyces coelicolor (O69809), Agrobacterium tumefaciens (Q44184), Geobacillus stearothermophilus (Q45515), Geobacillus stearothermophilus, Bacillus sp. (in: Bacteria) (Q56S49), Bacillus sp. (in: Bacteria), Pseudomonas putida (Q59699), Ralstonia pickettii (Q8VTT5), Bacillus sp. (in: Bacteria) TS-23 (Q56S49)
Werner, M.; Vasquez, F.J.; Fritz, C.; Vielhauer, O.; Siemann-Herzberg, M.; Altenbucher, J.; Syldatk, C.
Cloning of D-specific hydantoin utilization genes from Arthrobacter crystallopoietes
Eng. Life Sci.
4
563-572
2004
Geobacillus stearothermophilus (Q45515), Arthrobacter crystallopoietes (Q84FR6), Arthrobacter crystallopoietes DSM 20117 (Q84FR6)
-
Radha Kishan, K.V.; Vohra, R.M.; Ganesan, K.; Agrawal, V.; Sharma, V.M.; Sharma, R.
Molecular structure of D-hydantoinase from Bacillus sp. AR9: evidence for mercury inhibition
J. Mol. Biol.
347
95-105
2005
Paenarthrobacter aurescens, Geobacillus stearothermophilus, Bacillus sp. (in: Bacteria), Ralstonia pickettii, Thermus sp., Bacillus sp. (in: Bacteria) AR9
Lee, S.; Chang, Y.; Shin, D.; Han, J.; Seo, M.; Fazelinia, H.; Maranas, C.; Kim, H.
Designing the substrate specificity of D-hydantoinase using a rational approach
Enzyme Microb. Technol.
44
170-175
2009
Geobacillus stearothermophilus (Q45515)
-
cheon, Y.-H.; Kim, H.-S.; Han, K.-H.; Abendroth, J.; Niefind, K.; Schomburg, D.; Wang, J.; Kim, Y.
Crystal structure of D-hydantoinase from Bacillus stearothermophilus: insight into the stereochemistry of enantioselectivity
Biochemistry
41
9410-9417
2002
Geobacillus stearothermophilus (Q45515), Geobacillus stearothermophilus, Geobacillus stearothermophilus SD1 (Q45515)
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