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Literature summary for 3.5.1.23 extracted from

  • 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 (2009), J. Biol. Chem., 284, 9566-9577.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.5.1.23

Cloned(Commentary)

Cloned (Comment) Organism
for expression in human HEK-293 cells Rattus norvegicus
into the vector pET23a for expression in Escherichia coli BL21DE3 pLysS cells, in B834DE3 pLysS cells for preparation of selenomethionine-substituted enzyme Pseudomonas aeruginosa

Crystallization (Commentary)

Crystallization (Comment) Organism
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 Pseudomonas aeruginosa

Protein Variants

Protein Variants Comment Organism
E411A mutant, Zn2+-binding site mutated Pseudomonas aeruginosa
H177A mutant, deprotonation site mutated Rattus norvegicus
H97A mutant, Zn2+-binding site mutated Pseudomonas aeruginosa
H97A/H99A mutant, Zn2+-binding site and deprotonation site mutated Pseudomonas aeruginosa
H99A mutant, deprotonation site mutated Pseudomonas aeruginosa
R160A mutant, deprotonation site mutated Pseudomonas aeruginosa
R238A mutant, deprotonation site mutated Rattus norvegicus
Y448A mutant, Zn2+-binding site, deprotonation site mutated Pseudomonas aeruginosa
Y460A mutant, interaction site with ceramide mutated Pseudomonas aeruginosa
Y560A mutant, deprotonation site, Zn2+-binding site mutated Rattus norvegicus

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+
-
 Pseudomonas aeruginosa
Ca2+
-
 Rattus norvegicus
Mg2+
-
 Pseudomonas aeruginosa
Mg2+
-
 Rattus norvegicus
Zn2+
-
 Pseudomonas aeruginosa
Zn2+
-
 Rattus norvegicus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
additional information Pseudomonas aeruginosa ceramidase catalyzes the hydrolysis of ceramide to generate sphingosine and fatty acid, also found to catalyze the reverse reaction ?
-
 ?
additional information Rattus norvegicus ceramidase catalyzes the hydrolysis of ceramide to generate sphingosine and fatty acid, also found to catalyze the reverse reaction ?
-
 ?

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa Q9I596
-
-
Rattus norvegicus Q91XT9
-
-

Purification (Commentary)

Purification (Comment) Organism
using a Ni-Sepharose 6 Fast Flow and a Superdex 200 HR column Pseudomonas aeruginosa

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.00032
-
 mutant H97A/H99A Pseudomonas aeruginosa
0.00086
-
 mutant Y448A Pseudomonas aeruginosa
0.00198
-
 mutant H99A Pseudomonas aeruginosa
0.00228
-
 mutant R160A Pseudomonas aeruginosa
0.00301
-
 mutant H97A Pseudomonas aeruginosa
0.00844
-
 mutant H177A Rattus norvegicus
0.00944
-
 mutant R238A Rattus norvegicus
0.0104
-
 mutant Y560A Rattus norvegicus
0.0129
-
 wild type Rattus norvegicus
0.0146
-
 mutant Y460A Pseudomonas aeruginosa
0.02024
-
 wild type Pseudomonas aeruginosa
0.0387
-
 mutant E411A Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
C12-4-nitrobenzo-2-oxa-1,3-diazole-ceramide + H2O substrate activity assay Pseudomonas aeruginosa ?
-
 ?
C12-4-nitrobenzo-2-oxa-1,3-diazole-ceramide + H2O substrate activity assay Rattus norvegicus ?
-
 ?
additional information ceramidase catalyzes the hydrolysis of ceramide to generate sphingosine and fatty acid, also found to catalyze the reverse reaction Pseudomonas aeruginosa ?
-
 ?
additional information ceramidase catalyzes the hydrolysis of ceramide to generate sphingosine and fatty acid, also found to catalyze the reverse reaction Rattus norvegicus ?
-
 ?

Synonyms

Synonyms Comment Organism
CDase
-
 Pseudomonas aeruginosa
CDase
-
 Rattus norvegicus
N-acylsphingosine amidohydrolase
-
 Pseudomonas aeruginosa
N-acylsphingosine amidohydrolase
-
 Rattus norvegicus
neutral ceramidase
-
 Pseudomonas aeruginosa
neutral ceramidase
-
 Rattus norvegicus
PaCD
-
 Pseudomonas aeruginosa

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 activity assay Pseudomonas aeruginosa
37
-
 activity assay Rattus norvegicus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 activity assay Rattus norvegicus
8.5
-
 activity assay Pseudomonas aeruginosa