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Literature summary extracted from

  • Sumida, T.; Fujimoto, K.; Ito, M.
    Molecular cloning and catalytic mechanism of a novel glycosphingolipid-degrading beta-N-acetylgalactosaminidase from Paenibacillus sp. TS12 (2011), J. Biol. Chem., 286, 14065-14072.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.53 expression in Escherichia coli Paenibacillus sp.

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.53 D607E Km is increased by 5-fold, the kcat is decreased to 1.4% of wild-type Paenibacillus sp.
3.2.1.53 D607N activity is completely abolished Paenibacillus sp.
3.2.1.53 E608D Km value is decreased to about 57% and the kcat value is decreased to about 5.2% of wild-type Paenibacillus sp.
3.2.1.53 E608Q Km value is decreased to about 26% and the kcat value is decreased to about 1.1% of wild-type Paenibacillus sp.

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.2.1.53 Cu2+ 5 mM, strong inhibition Paenibacillus sp.
3.2.1.53 Hg2+ 5 mM, strong inhibition Paenibacillus sp.
3.2.1.53 N-acetylgalactosaminide thiazoline competitive Paenibacillus sp.
3.2.1.53 N-acetylglucosaminide thiazoline
-
 Paenibacillus sp.
3.2.1.53 Ni2+ 5 mM, strong inhibition Paenibacillus sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.53 0.09
-
 4-nitrophenyl beta-N-acetylgalactosaminide mutant E608Q, pH 6.0, 37°C Paenibacillus sp.
3.2.1.53 0.2
-
 4-nitrophenyl beta-N-acetylgalactosaminide mutant E608D, pH 6.0, 37°C Paenibacillus sp.
3.2.1.53 0.35
-
 4-nitrophenyl beta-N-acetylgalactosaminide pH 6.0, 37°C Paenibacillus sp.
3.2.1.53 1.74
-
 4-nitrophenyl beta-N-acetylgalactosaminide mutant D607E, pH 6.0, 37°C Paenibacillus sp.

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.53 115000
-
 x * 115000, SDS-PAGE Paenibacillus sp.

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.53 Paenibacillus sp. E9RH20
-
-

Reaction

EC Number Reaction Comment Organism Reaction ID
3.2.1.53 (GalNAcbeta(1-4))n + H2O = (GalNAcbeta(1-4))n-1 + 2-(acetylamino)-2-deoxy-beta-D-galactopyranose NgaP adopts substrate-assisted catalysis in which residues Glu608 and Asp607 could function as a proton donor and a stabilizer of the 2-acetamide group of the beta-GalNAc at the active site, respectively Paenibacillus sp.
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.53 4-nitrophenyl beta-N-acetylglucosaminide + H2O 1-3% of the activity with 4-nitrophenyl beta-N-acetylgalactosaminide Paenibacillus sp. beta-N-acetylglucosamine + 4-nitrophenol
-
 ?
3.2.1.53 4-nitrophenyl N-acetyl-beta-D-galactosaminopyranoside + H2O
-
 Paenibacillus sp. N-acetyl-D-galactosamine + 4-nitrophenol
-
 ?
3.2.1.53 GalNAc-beta-1,3-Gal-alpha-1,4-Gal-beta-1,4-Glc-beta-1,1-Cer + H2O i.e. globotetraosylceramide, 39% of the activity with GalNAc-1,4-Gal-1,4-Glc-1,1-Cer Paenibacillus sp. GalNAc + Gal-alpha-1,4-Gal-beta-1,4-Glc-beta-1,1-Cer
-
 ?
3.2.1.53 GalNAc-beta-1,4-Gal-beta-1,4-Glc-beta-1,1-Cer i.e. asialo-GM2 Paenibacillus sp. GalNAc + Gal-beta-1,4-Glc-beta-1,1-Cer
-
 ?
3.2.1.53 additional information enzyme cleaves the nonreducing terminal beta-GalNAc residues of gangliotriaosylceramide and globotetraosylceramide. Enzyme prefers the GalNAc-1,4-linkage over the GalNAc-1,3-linkage Paenibacillus sp. ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.53 ? x * 115000, SDS-PAGE Paenibacillus sp.

Synonyms

EC Number Synonyms Comment Organism
3.2.1.53 NgaP
-
 Paenibacillus sp.

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.53 0.08
-
 4-nitrophenyl beta-N-acetylgalactosaminide mutant E608Q, pH 6.0, 37°C Paenibacillus sp.
3.2.1.53 0.1
-
 4-nitrophenyl beta-N-acetylgalactosaminide mutant D607E, pH 6.0, 37°C Paenibacillus sp.
3.2.1.53 0.38
-
 4-nitrophenyl beta-N-acetylgalactosaminide mutant E608D, pH 6.0, 37°C Paenibacillus sp.
3.2.1.53 7.3
-
 4-nitrophenyl beta-N-acetylgalactosaminide wild-type, pH 6.0, 37°C Paenibacillus sp.

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
3.2.1.53 0.0000013
-
 N-acetylgalactosaminide thiazoline pH 6.0, 37°C Paenibacillus sp.
3.2.1.53 0.0468
-
 N-acetylglucosaminide thiazoline pH 6.0, 37°C Paenibacillus sp.

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.53 0.06
-
 4-nitrophenyl beta-N-acetylgalactosaminide mutant D607E, pH 6.0, 37°C Paenibacillus sp.
3.2.1.53 0.88
-
 4-nitrophenyl beta-N-acetylgalactosaminide mutant E608Q, pH 6.0, 37°C Paenibacillus sp.
3.2.1.53 1.9
-
 4-nitrophenyl beta-N-acetylgalactosaminide mutant E608D, pH 6.0, 37°C Paenibacillus sp.
3.2.1.53 20.9
-
 4-nitrophenyl beta-N-acetylgalactosaminide pH 6.0, 37°C Paenibacillus sp.