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Information on EC 5.1.99.5 - hydantoin racemase and Organism(s) Pseudomonas sp. and UniProt Accession Q00924

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EC Tree
     5 Isomerases
         5.1 Racemases and epimerases
             5.1.99 Acting on other compounds
                5.1.99.5 hydantoin racemase
IUBMB Comments
This enzyme, along with N-carbamoylase (EC 3.5.1.77 and EC 3.5.1.87) and hydantoinase, forms part of the reaction cascade known as the "hydantoinase process", which allows the total conversion of D,L-5-monosubstituted hydantoins into optically pure D- or L-amino acids . The enzyme from Pseudomonas sp. (HyuE) has a preference for hydantoins with aliphatic substituents, e.g. D- and L-5-[2-(methylsulfanyl)ethyl]hydantoin, whereas that from Arthrobacter aurescens shows highest activity with arylalkyl substituents, especially 5-benzylhydantoin, at the 5-position . In the enzyme from Sinorhizobium meliloti, Cys76 is responsible for recognition and proton retrieval of D-isomers, while Cys181 is responsible for L-isomer recognition and racemization .
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This record set is specific for:
Pseudomonas sp.
UNIPROT: Q00924
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Word Map
The taxonomic range for the selected organisms is: Pseudomonas sp.
The expected taxonomic range for this enzyme is: Bacteria, Archaea
Synonyms
hydantoin racemase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5'-monosubstituted-hydantoin racemase
-
-
-
-
hydantoin racemase
-
-
-
-
HyuA
-
-
-
-
HyuE
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
D-5-monosubstituted-hydantoin racemase
This enzyme, along with N-carbamoylase (EC 3.5.1.77 and EC 3.5.1.87) and hydantoinase, forms part of the reaction cascade known as the "hydantoinase process", which allows the total conversion of D,L-5-monosubstituted hydantoins into optically pure D- or L-amino acids [7]. The enzyme from Pseudomonas sp. (HyuE) has a preference for hydantoins with aliphatic substituents, e.g. D- and L-5-[2-(methylsulfanyl)ethyl]hydantoin, whereas that from Arthrobacter aurescens shows highest activity with arylalkyl substituents, especially 5-benzylhydantoin, at the 5-position [2]. In the enzyme from Sinorhizobium meliloti, Cys76 is responsible for recognition and proton retrieval of D-isomers, while Cys181 is responsible for L-isomer recognition and racemization [6].
CAS REGISTRY NUMBER
COMMENTARY hide
111310-51-1
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
D-5-isobutylhydantoin
L-5-isobutylhydantoin
show the reaction diagram
complete racemization
-
-
r
D-5-methylhydantoin
L-5-methylhydantoin
show the reaction diagram
complete racemization
-
-
r
L-5-(2-methylthioethyl)hydantoin
D-5-(2-methylthioethyl)hydantoin
show the reaction diagram
complete racemization
-
-
r
L-5-methylhydantoin
D-5-methylhydantoin
show the reaction diagram
about 50% racemization
-
-
r
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
slight stimulation
Mn2+
slight stimulation
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Cu2+
strong inhibition
DL-5-isopropylhydantoin
complete inhibition, pre-incubation with DL-5-(2-methylthioethyl)hydantoin protects
Zn2+
strong inhibition
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
29.8
D-5-(2-methylthioethyl)hydantoin
pH 7.5, 30°C
79.7
L-5-(2-methylthioethyl)hydantoin
pH 7.5, 30°C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
20.8
pH 7.5, 30°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
11
95% of maximum activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
UniProt
Manually annotated by BRENDA team
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
190000
gel filtration
32000
x * 32000, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
x * 32000, SDS-PAGE
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45
in presence of substrate, stable
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, ammonium sulfate at 90% saturation, stable for 6 months
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
-
stereospecific production of L-amino acids from the corresponding DL-5-substituted hydantoins. DL-5-substituted hydantoins are converted exclusively to the L-forms of the corresponding N-carbamyl-amino acids by hydantoinase in combination with hydantoin racemase. The N-carbamyl-l-amino acids are then converted to L-amino acids by N-carbamyl-L-amino acid amidohydrolase
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Ishikawa, T.; Watabe, K.; Mukohara, Y.; Nakamura, H.
Mechanism of stereospecific conversion of DL-5-substituted hydantoins to the corresponding L-amino acids by Pseudomonas sp. strain NS671
Biosci. Biotechnol. Biochem.
61
185-187
1997
Pseudomonas sp., Pseudomonas sp. NS671
Manually annotated by BRENDA team
Watabe, K.; Ishikawa, T.; Mukohara, Y.; Nakamura, H.
Purification and characterization of the hydantoin racemase of Pseudomonas sp. strain NS671 expressed in Escherichia coli
J. Bacteriol.
174
7989-7995
1992
Pseudomonas sp. (Q00924), Pseudomonas sp., Pseudomonas sp. NS671 (Q00924)
Manually annotated by BRENDA team