Information on EC 3.5.3.22 - proclavaminate amidinohydrolase

for references in articles please use BRENDA:EC3.5.3.22
Word Map on EC 3.5.3.22
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:


The expected taxonomic range for this enzyme is: Streptomyces clavuligerus

EC NUMBER
COMMENTARY hide
3.5.3.22
-
RECOMMENDED NAME
GeneOntology No.
proclavaminate amidinohydrolase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
amidinoproclavaminate + H2O = proclavaminate + urea
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of C-N bond
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
clavulanate biosynthesis
-
-
Clavulanic acid biosynthesis
-
-
Biosynthesis of antibiotics
-
-
SYSTEMATIC NAME
IUBMB Comments
amidinoproclavaminate amidinohydrolase
Forms part of the pathway for the biosythesis of the beta-lactamase inhibitor clavulanate in Streptomyces clavuligerus. It carries out an intermediary reaction between the first reaction of EC 1.14.11.21, clavaminate synthase, and the second and third reactions of that enzyme. Requires Mn2+.
CAS REGISTRY NUMBER
COMMENTARY hide
9000-96-8
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
metabolism
the enzyme is involved in clavulanic acid biosynthesis
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
(3R)-hydroxy-N-acetyl-L-arginine + H2O
?
show the reaction diagram
-
-
-
-
?
deoxyguanidinoproclavaminic acid + H2O
deoxyproclavaminic acid + urea
show the reaction diagram
guanidinoproclavaminic acid + H2O
proclavaminic acid + urea
show the reaction diagram
N-acetyl-L-arginine + H2O
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
guanidinoproclavaminic acid + H2O
proclavaminic acid + urea
show the reaction diagram
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
-
preferred metal activator
Ni2+
-
50% of the activity with Co2+ as activator
additional information
-
no activation by Ca2+, Fe2+ and Zn2+
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
12
(3R)-hydroxy-N-acetyl-L-arginine
-
pH 9.0, 37°C
30
N-acetyl-L-arginine
-
pH 9.0, 37°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
33000
-
x * 33000, SDS-PAGE
33300
-
x * 33300, deduced from gene sequence
33401
-
6 * 33401
34000
-
x * 34000, SDS-PAGE
199500
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
-
6 * 33401
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging-drop vapour-diffusion method, crystal structures of the enzyme at 1.75 A and 2.45 A resolution. Crystals belong to space group P2(1), with unit cell dimensions a = 95.4 A, b = 81.9 A, c = 118.6 A beta = 95.5°, but following transfer into the sucrose solution they belong to C2, with unit cell dimensions a = 140.0 A, b = 79.0 A, c = 93.5 A and beta = 124.0°
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant enzyme
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
amplified from Streptomyces clavuligerus genomic DNA. Cloned in the pSET152 integration and pIBR25 expression vectors containing the strong ermE promoter. Plasmids are introduced into Streptomyces clavuligerus by protoplast transformation
-
expressed in Streptomyces clavuligerus strain SMF5704
-
Streptomyces venezuelae YJ028 is utilized as the heterologous host for the expression of four structural clavulanic acid biosynthesis genes, which encode carboxyethylarginine synthase (ceas2), beta-lactam synthetase (bls2), clavaminate synthase (cas2), and proclavaminate amidinohydrolase (pah2). The genes are cloned into pIBR25 expression vector containing ermE promoter to generate pBS4. The cas2 gene is also cloned into pSET152 to generate pCas2. It is then integrated into the genome of Streptomyces venezuelae YJ028
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
transcription of pah2 is increased in transformants over-expressing pah2
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
overexpression as fusion to maltose-binding protein
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
synthesis
clavulanic acid intermediates: deoxygaunidinoproclavaminic acid, guanidinoproclavaminic acid, and dihydroclavaminic acid are heterologously produced in Streptomyces venezuelae recombinant using four sets of early genes from the clavulanic acid biosynthetic pathway