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Information on EC 3.5.1.23 - ceramidase and Organism(s) Pseudomonas aeruginosa and UniProt Accession Q9I596
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3.5.1.23 ceramidaseSpecify your search results
Word Map
- 3.5.1.23
-
adipor1
- sphingolipids
- obesity
-
adipokine
- agonist
- sphingomyelinase
- sphingomyelin
- lysosomal
- farber
-
obese
- leptin
- adipocytes
-
amp-activated
- sphingosine-1-phosphate
- peroxisome
- cardiovascular
-
proliferator-activated
- triglyceride
-
globular
-
insulin-sensitizing
-
high-fat
- tnf
- smases
-
adipocytokine
-
adipocyte-derived
-
monophosphate-activated
-
anti-atherogenic
-
t-cadherin
- steatosis
-
adiposity
- glucosylceramide
- glycosphingolipids
-
obesity-related
-
antidiabetic
- 1-phosphate
-
seven-transmembrane
- gaucher
- dihydrosphingosine
-
p-ampk
- palmitoyltransferase
-
homa-ir
-
sphingoid
- hoarseness
-
ceramide-induced
- sphinganine
-
ceramide-mediated
- dihydroceramide
-
myoclonic
-
adipocyte-secreted
- analysis
- pharmacology
- drug development
- medicine
-
hypoadiponectinemia
The taxonomic range for the selected organisms is: Pseudomonas aeruginosa
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
The expected taxonomic range for this enzyme is: Eukaryota, Bacteria, Archaea
Synonyms
adipor2, adiponectin receptor, ceramidase, acid ceramidase, asah1, neutral ceramidase, acdase, ncdase, acer2, acer3, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
acylsphingosine deacylase
-
-
-
-
glycosphingolipid ceramide deacylase
-
-
-
-
N-acylsphingosine amidohydrolase
-
-
-
-
PHP32
-
-
-
-
Putative 32 KDA heart protein
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a ceramide + H2O = a carboxylate + sphingosine
neutral CDase contains a zinc ion in the active site that functions as a catalytic center, and the hydrolysis of the N-acyl linkage in ceramide proceeds through a mechanism that is similar to that described for zinc-dependent carboxypeptidase, reaction mechanism, overview
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
hydrolysis of peptide bond
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
N-acylsphingosine amidohydrolase
-
CAS REGISTRY NUMBER
COMMENTARY
37289-06-8
-
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
C12-4-nitrobenzo-2-oxa-1,3-diazole-ceramide + H2O
?
substrate activity assay
-
-
?
N-acylsphingosine + H2O
a carboxylate + sphingosine
CDase hydrolyzes the amide bond in ceramides to yield free fatty acid and sphingosine
-
-
?
N-palmitoyl sphingosine + H2O
palmitic acid + sphingosine
-
-
-
?
(4E,2S,3R)-2-N-(10-pyrenedecanoyl)-1,3,17-trihydroxy-17-(3,5-dinitrobenzoyl)-4-heptadecene + H2O
?
-
-
fluorescent product formed upon hydrolysis of the N-acyl bond
-
?
4-nitrobenzo-2-oxa-1,3-diazole-ceramide + H2O
?
-
C12 substrate chain length
-
-
?
D-erythro-sphingosine + (4Z,7Z,10Z,13Z,16Z,19Z)-docosahexa-4,7,10,13,16,19-enoic acid
N-((4Z,7Z,10Z,13Z,16Z,19Z)-docosahexa-4,7,10,13,16,19-enoyl)-D-erythro-sphingosine + H2O
-
50% of activity
-
-
r
D-erythro-sphingosine + arachidonic acid
N-arachidonyl-D-erythro-sphingosine + H2O
-
61% of activity
-
-
r
D-erythro-sphingosine + behenic acid
N-behenoyl-D-erythro-sphingosine + H2O
-
24% of activity
-
-
r
D-erythro-sphingosine + cerotic acid
N-cerotoyl-D-erythro-sphingosine + H2O
-
4% of activity
-
-
r
D-erythro-sphingosine + eicosapentaenoic acid
N-eicosapentaenyl-D-erythro-sphingosine + H2O
-
52% of activity
-
-
r
D-erythro-sphingosine + fatty acid
N-acyl-D-erythro-sphingosine + H2O
-
-
-
-
r
D-erythro-sphingosine + lauric acid
N-lauroyl-D-erythro-sphingosine + H2O
-
72% of activity
-
-
r
D-erythro-sphingosine + lignoceric acid
N-lignoceroyl-D-erythro-sphingosine + H2O
-
6% of activity
-
-
r
D-erythro-sphingosine + linoleic acid
N-linoleoyl-D-erythro-sphingosine + H2O
-
66% of activity
-
-
r
D-erythro-sphingosine + myristic acid
N-myristoyl-D-erythro-sphingosine + H2O
-
best substrate
-
-
r
D-erythro-sphingosine + oleic acid
N-oleoyl-D-erythro-sphingosine + H2O
-
39% of activity
-
-
r
D-erythro-sphingosine + palmitic acid
N-palmitoyl-D-erythro-sphingosine + H2O
-
70% of activity, palmitic acid can not be replaced with palmityl-CoA
-
-
r
D-erythro-sphingosine + stearic acid
N-stearoyl-D-erythro-sphingosine + H2O
-
49% of activity
-
-
r
N-acylsphingosine + H2O
a carboxylate + sphingosine
-
CDase cleaves the N-acyl linkage of ceramide to produce sphingosine and free fatty acid
-
-
?
N-acylsphingosine + H2O
a carboxylate + sphingosine
-
involved in the biosynthesis of ceramide, crucial enzyme for the regulation of the intracellular balance of the contents of ceramide, sphingosine and possibly sphingosine 1-phosphate
-
-
?
N-acylsphingosine + H2O
a carboxylate + sphingosine
-
involved in the biosynthesis of ceramide, crucial enzyme for the regulation of the intracellular balance of the contents of ceramide, sphingosine and possibly sphingosine 1-phosphate
-
-
r
additional information
?
-
CDase may be a virulence factor
-
-
?
additional information
?
-
ceramidase catalyzes the hydrolysis of ceramide to generate sphingosine and fatty acid, also found to catalyze the reverse reaction
-
-
?
additional information
?
-
-
ceramidase catalyzes the hydrolysis of ceramide to generate sphingosine and fatty acid, also found to catalyze the reverse reaction
-
-
?
additional information
?
-
-
the C-terminal tail is essential for enzyme activity and correct folding
-
-
?
additional information
?
-
-
ceramidase enhances phospholipase C-induced hemolysis by Pseudomonas aeruginosa
-
-
?
additional information
?
-
-
ceramidase enhances cytotoxicity induced by hemolytic phospholipase C
-
-
?
additional information
?
-
-
the Pseudomonas neutral CDase can hydrolyze ceramide in intact erythrocytes, leading to hemolysis. The reaction mechanism for ceramide hydrolysis in vivo appears to be the same as that for ceramide hydrolysis in vitro because the mutation of residues surrounding the catabolic zinc ion abolish the hemolytic activity along with ceramide hydrolysis by CDase in intact erythrocytes
-
-
?
additional information
?
-
-
the enzyme can hydrolyze human skin-specific omega-hydroxyacyl ceramides
-
-
?
additional information
?
-
-
the enzyme is able to effectively degrade ceramide in the presence of Staphylococcus aureus-derived lipids or neutral detergents
-
-
?
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
N-acylsphingosine + H2O
a carboxylate + sphingosine
-
involved in the biosynthesis of ceramide, crucial enzyme for the regulation of the intracellular balance of the contents of ceramide, sphingosine and possibly sphingosine 1-phosphate
-
-
?
N-acylsphingosine + H2O
a carboxylate + sphingosine
-
involved in the biosynthesis of ceramide, crucial enzyme for the regulation of the intracellular balance of the contents of ceramide, sphingosine and possibly sphingosine 1-phosphate
-
-
r
additional information
?
-
CDase may be a virulence factor
-
-
?
additional information
?
-
ceramidase catalyzes the hydrolysis of ceramide to generate sphingosine and fatty acid, also found to catalyze the reverse reaction
-
-
?
additional information
?
-
-
ceramidase catalyzes the hydrolysis of ceramide to generate sphingosine and fatty acid, also found to catalyze the reverse reaction
-
-
?
additional information
?
-
-
ceramidase enhances phospholipase C-induced hemolysis by Pseudomonas aeruginosa
-
-
?
additional information
?
-
-
ceramidase enhances cytotoxicity induced by hemolytic phospholipase C
-
-
?
additional information
?
-
-
the Pseudomonas neutral CDase can hydrolyze ceramide in intact erythrocytes, leading to hemolysis. The reaction mechanism for ceramide hydrolysis in vivo appears to be the same as that for ceramide hydrolysis in vitro because the mutation of residues surrounding the catabolic zinc ion abolish the hemolytic activity along with ceramide hydrolysis by CDase in intact erythrocytes
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Ca2+
Mg2+
Ca2+
Mg2+
Zn2+
-
neutral CDase contains a zinc ion in the active site that functions as a catalytic center, the active center of CDase is the zinc ion itself, with the reaction following a similar mechanism as observed for zinc-dependent carboxypeptidase
additional information
-
divalent cation required, without strong specificity
Mg2+
the maximum activity with Mg2+ is only 36% of that in presence of optimal Ca2+ or Ba2+ concentrations
Ca2+
-
a calcium or magnesium ion is presumed to be important for stabilizing the two domains
Mg2+
-
a calcium or magnesium ion is presumed to be important for stabilizing the two domains
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
D-erythro-sphingosine
recombinant CDase, more inhibitory than L-erythro-sphingosine
EDTA
recombinant CDase, complete inhibition, Ca2+ restores activity
EGTA
recombinant CDase, complete inhibition, Ca2+ restores activity
L-erythro-sphingosine
recombinant CDase, 0.02 mM, 20% inhibition, less inhibitory than L-erythro-sphingosine
1-ethyl-3-(3-dimethylaminopropyl)carbodiimide
-
modification of carboxyl groups completely inhibits forward and reverse reaction
2,3-Butanedione
-
modification of Arg causes about 50% inhibition of forward and reverse reaction
N-bromosuccinimide
-
modification of Trp completely inhibits forward and reverse reaction
additional information
not inhibited by 1 mM iodoacetic acid, 0.1 mM 2-mercaptoethanol, 0.05 mM hexadecylsulfonylfluoride, 1 mM p-phenylmethylsulfonylfluoride, 0.2 mM D-erythro-2-(N-myristoylamino)-1-phenyl-1-propanol, 0.5 mM N-oleoylethanolamine, L(-)- and D(-)-norephenedrin
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
cardiolipin
-
phosphatidylglycerol and cardiolipin, which are major lipid components of Staphylococcus aureus, are effective in stimulating the hydrolysis of human skin-type ceramides in the absence of detergents
phosphatidylglycerol
-
phosphatidylglycerol and cardiolipin, which are major lipid components of Staphylococcus aureus, are effective in stimulating the hydrolysis of human skin-type ceramides in the absence of detergents
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.0005
(4E,2S,3R)-2-N-(10-pyrenedecanoyl)-1,3,17-trihydroxy-17-(3,5-dinitrobenzoyl)-4-heptadecene
-
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.1
(4E,2S,3R)-2-N-(10-pyrenedecanoyl)-1,3,17-trihydroxy-17-(3,5-dinitrobenzoyl)-4-heptadecene
-
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.218
intracellular recombinant CDase expressed in Escherichia coli DH5alpha, pH 8.5, hydrolysis of N-palmitoyl sphingosine
0.239
extracellular wild-type CDase, pH 8.5, hydrolysis of N-palmitoyl sphingosine
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
8.5
8.5
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
4.9
recombinant CDase from inclusion bodies, expressed in Escherichia coli BL21 without its signal peptide, isoelectric focusing
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SOURCE
-
when the strain is cultured with sphingomyelin or phosphatidylcholine, production of the enzyme drastically increases, causing the increase of hemolytic activity in the cellfree culture supernatant. Ceramide and sphingosine are effective in promoting the production of ceramidase
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
wild-type CDase from 3-L culture, recombinant CDase expressed in Pseudomonas putida KT2440 and Pichia pastoris GS115
-
inactive recombinant CDase expressed in Escherichia coli BL21 without its signal peptide
recombinant CDase expressed in Escherichia coli DH5alpha and BL21
recombinant CDase expressed in Escherichia coli DH5alpha is most likely present in periplasm
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
-
ceramidases are classified into three distinct groups, acid (Asah1), neutral (Asah2), and alkaline (Asah3) CDases, based on their primary structure and optimum pH. Acid CDase catabolizes ceramide in lysosomes and is found only in vertebrates. In contrast, the distribution of neutral and alkaline CDases is broad, with both being found in species ranging from lower eukaryotes to mammals; however, only neutral CDase is found in prokaryotes, including some pathogenic bacteria. Neutral CDase is thought to have gained a specific domain (mucin box) in the N-terminal region after the vertebrate split, allowing the enzyme to be stably expressed at the plasmamembrane as a type II membrane protein. Molecular evolution of neutral ceramidase acquiring a mucin box, overview
physiological function
-
the bacterial enzyme alone does not affect TNF-alpha gene expression in three-dimensionally cultured human primary keratinocytes. In the presence of the detergent Triton X-100, which damages stratum corneum structure, the active enzyme, but not heat-inactivated enzyme or inactive mutant enzyme, induces the production of TNF-alpha, endothelin-1, and interleukin-8, indicating that this production is dependent on ceramidase activity, it is also inhibited by a sphingosine kinase inhibitor and by an sphingosine 1-phosphate receptor antagonist VPC 23019. Among various ceramide metabolites, sphingosine and sphingosine 1-phosphate enhance the gene expression of TNF-alpha, endothelin-1, and interleukin-8, overview
additional information
-
enzyme structure-function relationship, overview. The enzyme contains a signal/anchor sequence but no mucin box
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
additional information
-
the C-terminal tail is essential for enzyme activity and correct folding
additional information
-
neutral CDase is composed of two domains: a novel NH2-terminal domain harboring an active site and an immunoglobulin-like COOH-terminal domain. A zinc-binding site is located in the center of the NH2-terminal domain, whereas a calcium/magnesium-binding site is found at the interface between the NH2-terminal and COOH-terminal domains
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
glycoprotein
recombinant CDase expressed in Pichia pastoris GS115
additional information
CDase contains a 26 amino acids signal peptide
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
the crystal structures of neutral CDase from Pseudomonas aeruginosa in the C2-ceramide-bound and -unbound forms are determined at 2.2 and 1.4 A resolutions, respectively
crystal structure of neutral CDase for the ceramide-free and C2-ceramide-bound forms
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
H97A/H99A
mutant, Zn2+-binding site and deprotonation site mutated
additional information
additional information
-
truncation of six C-terminal amino acids, but not of five, results in complete loss of CDase activity
additional information
-
construction of a CDase-null mutant via homologous recombination and gene disruption, CDase-deficient mutants lack any CDase activity, the hemolytic activity by the CDase mutants against human erythrocytes is significantly reduced compared with that by the wild-type
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-85°C, 2 months, with 1 mg/ml bovine serum albumin, no loss of activity
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
wild-type CDase from 3-L culture: 217fold, recombinant CDase expressed in Escherichia coli
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression in Escherichia coli DH5alpha and BL21, expression at low levels in Pseudomonas putida KT2440 and Pichia pastoris GS115, sequencing
into the vector pET23a for expression in Escherichia coli BL21DE3 pLysS cells, in B834DE3 pLysS cells for preparation of selenomethionine-substituted enzyme
gene cdase, the gene encoding the enzyme is adjacent to that encoding hemolytic phospholipase C, plcH, in the genome of Pseudomonas aeruginosa
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
CDase from Pseudomonas aeruginosa may be a virulence factor in infections, may be involved in the ceramide deficiency in the horny layer of the epidermis of patients with atopic dermatitis suffering from bacterial infections
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Okino, N.; Tani, M.; Imayama, S.; Ito, M.
Purification and characterization of a novel ceramidase from Pseudomonas aeruginoa
J. Biol. Chem.
273
14368-14373
1998
Pseudomonas aeruginosa
Tani, M.; Okino, N.; Sueyoshi, N.; Ito, M.
Conserved amino acid residues in the COOH-terminal tail are indispensable for the correct folding and localization and enzyme activity of neutral ceramidase
J. Biol. Chem.
279
29351-29358
2004
Pseudomonas aeruginosa, Rattus norvegicus
Nieuwenhuizen, W.F.; van Leeuwen, S.; Jack, R.W.; Egmond, M.R.; Gotz, F.
Molecular cloning and characterization of the alkaline ceramidase from Pseudomonas aeruginosa PA01
Protein Expr. Purif.
30
94-104
2003
Pseudomonas aeruginosa (Q9I596)
Kita, K.; Okino, N.; Ito, M.
Reverse hydrolysis reaction of a recombinant alkaline ceramidase of Pseudomonas aeruginosa
Biochim. Biophys. Acta
1485
111-120
2000
Pseudomonas aeruginosa
Nieuwenhuizen, W.F.; van Leeuwen, S.; Gotz, F.; Egmond, M.R.
Synthesis of a novel fluorescent ceramide analogue and its use in the characterization of recombinant ceramidase from Pseudomonas aeruginosa PA01
Chem. Phys. Lipids
114
181-191
2002
Pseudomonas aeruginosa
Okino, N.; Ito, M.
Ceramidase enhances phospholipase C-induced hemolysis by Pseudomonas aeruginosa
J. Biol. Chem.
282
6021-6030
2007
Pseudomonas aeruginosa, Pseudomonas aeruginosa AN17
Inoue, T.; Okino, N.; Kakuta, Y.; Hijikata, A.; Okano, H.; Goda, H.M.; Tani, M.; Sueyoshi, N.; Kambayashi, K.; Matsumura, H.; Kai, Y.; Ito, M.
Mechanistic insights into the hydrolysis and synthesis of ceramide by neutral ceramidase
J. Biol. Chem.
284
9566-9577
2009
Rattus norvegicus (Q91XT9), Pseudomonas aeruginosa (Q9I596), Pseudomonas aeruginosa
Ito, M.; Okino, N.; Tani, M.
New insight into the structure, reaction mechanism, and biological functions of neutral ceramidase
Biochim. Biophys. Acta
1841
682-691
2014
Dictyostelium discoideum, Mycobacterium tuberculosis, Oryza sativa, Pseudomonas aeruginosa, Tribolium castaneum, Dermatophilus congolensis, Triticum aestivum (A9YFM2), Aspergillus oryzae (Q5B5D5), Danio rerio (Q5W7F1), Rattus norvegicus (Q91XT9), Mus musculus (Q9JHE3), Homo sapiens (Q9NR71), Drosophila melanogaster (Q9VA70), Laodelphax striatellus (R4N4U2)
Oizumi, A.; Nakayama, H.; Okino, N.; Iwahara, C.; Kina, K.; Matsumoto, R.; Ogawa, H.; Takamori, K.; Ito, M.; Suga, Y.; Iwabuchi, K.
Pseudomonas-derived ceramidase induces production of inflammatory mediators from human keratinocytes via sphingosine-1-phosphate
PLoS ONE
9
e89402
2014
Pseudomonas aeruginosa
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