Literature summary for 3.5.1.23 extracted from

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 (2003), Protein Expr. Purif., 30, 94-104.

Search for more literature for 3.5.1.23

Application

Application	Comment	Organism
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	Pseudomonas aeruginosa

Cloned(Commentary)

Cloned (Comment)	Organism
expression in Escherichia coli DH5alpha and BL21, expression at low levels in Pseudomonas putida KT2440 and Pichia pastoris GS115, sequencing	Pseudomonas aeruginosa

Inhibitors

Inhibitors	Comment	Organism	Structure
D-erythro-sphingosine	recombinant CDase, more inhibitory than L-erythro-sphingosine	Pseudomonas aeruginosa
DL-sphinganine	recombinant CDase	Pseudomonas aeruginosa
EDTA	recombinant CDase, complete inhibition, Ca2+ restores activity	Pseudomonas aeruginosa
EGTA	recombinant CDase, complete inhibition, Ca2+ restores activity	Pseudomonas aeruginosa
L-erythro-sphingosine	recombinant CDase, 0.02 mM, 20% inhibition, less inhibitory than L-erythro-sphingosine	Pseudomonas aeruginosa
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	Pseudomonas aeruginosa
phytosphingosine	recombinant CDase, 0.02 mM, 60% inhibition	Pseudomonas aeruginosa
sphingosine	strong inhibitor	Pseudomonas aeruginosa

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
extracellular	wild-type CDase from 3-L culture, recombinant CDase expressed in Pseudomonas putida KT2440 and Pichia pastoris GS115	Pseudomonas aeruginosa	-	-
inclusion body	inactive recombinant CDase expressed in Escherichia coli BL21 without its signal peptide	Pseudomonas aeruginosa	16234	-
intracellular	recombinant CDase expressed in Escherichia coli DH5alpha and BL21	Pseudomonas aeruginosa	5622	-
periplasm	recombinant CDase expressed in Escherichia coli DH5alpha is most likely present in periplasm	Pseudomonas aeruginosa	-	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Ba2+	better activator than Mg2+	Pseudomonas aeruginosa
Ca2+	Ca2+-dependent, better activator than Mg2+	Pseudomonas aeruginosa
Mg2+	the maximum activity with Mg2+ is only 36% of that in presence of optimal Ca2+ or Ba2+ concentrations	Pseudomonas aeruginosa

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
70000	-	x * 70000, wild-type and recombinant CDase, SDS-PAGE	Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
additional information	Pseudomonas aeruginosa	CDase may be a virulence factor	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas aeruginosa	Q9I596	strain PA01	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
glycoprotein	recombinant CDase expressed in Pichia pastoris GS115	Pseudomonas aeruginosa
additional information	CDase contains a 26 amino acids signal peptide	Pseudomonas aeruginosa

Purification (Commentary)

Purification (Comment)	Organism
wild-type CDase from 3-L culture: 217fold, recombinant CDase expressed in Escherichia coli	Pseudomonas aeruginosa

Source Tissue

Source Tissue	Comment	Organism	Textmining
cell culture	3-L culture	Pseudomonas aeruginosa	-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg]	Specific Activity Maximum [µmol/min/mg]	Comment	Organism
0.218	-	intracellular recombinant CDase expressed in Escherichia coli DH5alpha, pH 8.5, hydrolysis of N-palmitoyl sphingosine	Pseudomonas aeruginosa
0.239	-	extracellular wild-type CDase, pH 8.5, hydrolysis of N-palmitoyl sphingosine	Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
additional information	CDase may be a virulence factor	Pseudomonas aeruginosa	?	-	?
N-acylsphingosine + H2O	CDase hydrolyzes the amide bond in ceramides to yield free fatty acid and sphingosine	Pseudomonas aeruginosa	a carboxylate + sphingosine	-	?
N-palmitoyl sphingosine + H2O	-	Pseudomonas aeruginosa	palmitic acid + sphingosine	-	?

Subunits

Subunits	Comment	Organism
?	x * 70000, wild-type and recombinant CDase, SDS-PAGE	Pseudomonas aeruginosa

Synonyms

Synonyms	Comment	Organism
alkaline ceramidase	-	Pseudomonas aeruginosa
CDase	-	Pseudomonas aeruginosa

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8.5	-	wild-type and recombinant CDase	Pseudomonas aeruginosa

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.0033	-	DL-sphinganine	pH 8.5, recombinant CDase	Pseudomonas aeruginosa
0.004	-	D-erythro-sphingosine	pH 8.5, recombinant CDase	Pseudomonas aeruginosa

pI Value

Organism	Comment	pI Value Maximum	pI Value
Pseudomonas aeruginosa	recombinant CDase from inclusion bodies, expressed in Escherichia coli BL21 without its signal peptide, isoelectric focusing	-	4.9
Pseudomonas aeruginosa	sequence calculation	-	6

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Release 2023.1 (January 2023)