Any feedback?
Please rate this page
(enzyme.php)
(0/150)

BRENDA support

BRENDA Home
show all | hide all No of entries

Information on EC 3.5.2.10 - creatininase and Organism(s) Pseudomonas putida and UniProt Accession P83772

for references in articles please use BRENDA:EC3.5.2.10
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
     3 Hydrolases
         3.5 Acting on carbon-nitrogen bonds, other than peptide bonds
             3.5.2 In cyclic amides
                3.5.2.10 creatininase
Specify your search results
Select one or more organisms in this record: ?
This record set is specific for:
Pseudomonas putida
UNIPROT: P83772 not found.
Show additional data
Do not include text mining results
Include (text mining) results
Include results (AMENDA + additional results, but less precise)
Word Map
The taxonomic range for the selected organisms is: Pseudomonas putida
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Reaction Schemes
Synonyms
creatininase, creatinine amidohydrolase, creatinine hydrolase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
creatinine amidohydrolase
-
-
creatinine hydrolase
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
creatinine + H2O = creatine
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of peptide bond
-
-
-
-
PATHWAY SOURCE
PATHWAYS
-
-
SYSTEMATIC NAME
IUBMB Comments
creatinine amidohydrolase
-
CAS REGISTRY NUMBER
COMMENTARY hide
9025-13-2
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
creatinine + H2O
creatine
show the reaction diagram
-
-
-
?
creatinine + H2O
?
show the reaction diagram
-
-
-
-
?
creatinine + H2O
creatine
show the reaction diagram
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
creatinine + H2O
?
show the reaction diagram
-
-
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Mn2+
may substitute for Zn2+
Zinc
removal of the zinc ions by dialyzing against buffers containing o-phenanthroline results in a partial decrease of 10-30% in activity and zinc content. Wild-type enzyme, purified in the presence of EDTA, shows only half the activity of the wild-type enzyme, the activity is recovered by the addition of ZnCl2 or MnCl2
Co2+
-
activates
Fe2+
-
activates
Mg2+
-
activates
Mn2+
-
activates
Ni2+
-
activates
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
EDTA
wild-type enzyme, purified in the presence of EDTA, shows only half the activity of the wild-type enzyme, the activity is recovered by the addition of ZnCl2 or MnCl2
ethoxyformic anhydride
-
-
heavy metal ions
-
-
-
N-bromosuccinimide
-
-
o-phenanthroline
-
-
Photooxidation
-
-
-
additional information
-
-
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
44 - 350
creatinine
26
creatinine
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.5 - 1340
creatinine
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.008 - 17.4
creatinine
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.13
mutant E122Q, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication
13.9
mutant E122Q, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication
140
mutant Y121A, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication
1560
wild-type, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication
178
wild-type, presence of EDTA, semi-apo-enzyme, pH not specified in the publication, temperature not specified in the publication
370
wild-type, presence of Zn2+, pH not specified in the publication, temperature not specified in the publication
4.9
mutant W154F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication
740
mutant W174F, presence of Mn2+, pH not specified in the publication, temperature not specified in the publication
additional information
-
by Escherichia coli transformants pCRN741 without and with IPTG 0.678 U per ml
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
CRNA_PSEPU
260
0
28569
Swiss-Prot
-
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
170400
-
SDS-PAGE
173000
-
dynamic light scattering
175000
-
ultracentrifugal analysis
224000
-
gel filtration
23000
-
8 * 23000, SDS-PAGE
28399
-
6 * 28399, calculated from amino acid sequence
28400
-
6 * 28400, MALDI-TOF mass spectrometry
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
homohexamer
octamer
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
-
3.5% glucose
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystal structure of the enzyme complexed with a 1-methylguanidine. All subunits in the complex exist as the closed form. X-ray crystallographic study and the atomic absorption spectrometry analysis of the E122Q mutant-substrate complex reveal that the drastic decrease of the activity of the E122Q is caused by the loss of one Zn ion at the Metal1 site and a critical function of Glu122
crystal structures of the native and the Mn2+-activated enzyme determined by a difference Fourier method, crystal structure of Mn2+-activated enzyme determined by the single isomorphous replacement method
hanging drop vapour diffusion method with 1.5 M lithium sulfate in 0.1 M Na-HEPES pH 7.5
-
sitting drop vapour diffusion method with 22% ethanol, 20% PEG 8000, and 0.1 M MOPS pH 7.5
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E122Q
almost complete loss of activity
E183Q
complete loss of activity
H178A
complete loss of activity
W154A
complete loss of activity
W154F
almost complete loss of activity
W174A
almost complete loss of activity
W174F
about 50% of wild-type activity
Y121A
about 10% of wild-type catalytic efficiency
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 10
-
-
172080
6 - 12
-
for 3 h at 30°C
172079
6 - 9
-
the enzyme tends to precipitate at pH values of under 6.0 and over 9.0 corresponding to the stability range of the enzyme
677306
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
55
-
30 min at pH 7.1, apoenzyme
70
-
30 min at pH 7.1, native enzyme
75
-
30 min, activity still 40%
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
unstable to photooxidation
-
172079
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 6 months
-
4°C, 5 days
-
lyophilized
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
native and Mn2+-activated enzymes purified, Toyopearl HW65C column and DEAE-Toyopearl column
ammonium sulfate fractionation, DEAE Sepharose CL-6B and Sepharose CL-6B column chromatographies, 50% total activity
-
purified by column chromarographies on sarcosine-HM-Sepharose and DEAE-cellulose, and gel filtration on Sephadex G-200
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli
cloned, sequenced and expressed in Escherichia coli DH5?, vector pCRN741, creatininase gene consists of 771 bp, encodes 257 amino acids, similarity of RS65 to PS-7 72.8% and Arthrobacter sp. TE1826 43.2%
-
expressed in Escherichia coli
-
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
wild-type enzyme, purified in the presence of EDTA, shows only half the activity of the wild-type enzyme, the activity is recovered by the addition of ZnCl2 or MnCl2
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
analysis of serum creatinine concentration
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Tang, T.Y.; Wen, C.J.
Expression of the creatininase gene from Pseudomonas putida RS65 in Escherichia coli
J. Ind. Microbiol.
24
2-6
2000
Alcaligenes sp., Pseudomonas putida
-
Manually annotated by BRENDA team
Rikitake, K.; Oka, I.; Ando, M.; Yoshimoto, T.; Tsuru, D.
Creatinine amidohydrolase (creatininase) from Pseudomonas putida. Purification and some properties
J. Biochem.
86
1109-1117
1979
Pseudomonas putida
Manually annotated by BRENDA team
Tsuru, D.; Oka, I.; Yoshimoto, T.
Creatinine decomposing enzymes in Pseudomonas putida
Agric. Biol. Chem.
40
1011-1018
1976
Paenarthrobacter ureafaciens, Pseudomonas aeruginosa, Pseudomonas putida
-
Manually annotated by BRENDA team
Beuth, B.; Niefind, K.; Schomburg, D.
Crystal structure of creatininase from Pseudomonas putida: a novel fold and a case of convergent evolution
J. Mol. Biol.
332
287-301
2003
Pseudomonas putida (P83772), Pseudomonas putida
Manually annotated by BRENDA team
Yoshimoto, T.; Tanaka, N.; Kanada, N.; Inoue, T.; Nakajima, Y.; Haratake, M.; Nakamura, K.T.; Xu, Y.; Ito, K.
Crystal structures of creatininase reveal the substrate binding site and provide an insight into the catalytic mechanism
J. Mol. Biol.
337
399-416
2004
Pseudomonas putida (P83772), Pseudomonas putida
Manually annotated by BRENDA team
Beuth, B.; Niefind, K.; Schomburg, D.
Crystallization and preliminary crystallographic analysis of creatininase from Pseudomonas putida
Acta Crystallogr. Sect. D
58
1356-1358
2002
Pseudomonas putida
Manually annotated by BRENDA team
Ito, K.; Kanada, N.; Inoue, T.; Furukawa, K.; Yamashita, K.; Tanaka, N.; Nakamura, K.T.; Nishiya, Y.; Sogabe, A.; Yoshimoto, T.
Preliminary crystallographic studies of the creatinine amidohydrolase from Pseudomonas putida
Acta Crystallogr. Sect. D
58
2180-2181
2002
Pseudomonas putida
Manually annotated by BRENDA team
Yamashita, K.; Nakajima, Y.; Matsushita, H.; Nishiya, Y.; Yamazawa, R.; Wu, Y.F.; Matsubara, F.; Oyama, H.; Ito, K.; Yoshimoto, T.
Substitution of Glu122 by glutamine revealed the function of the second water molecule as a proton donor in the binuclear metal enzyme creatininase
J. Mol. Biol.
396
1081-1096
2010
Pseudomonas putida (P83772)
Manually annotated by BRENDA team