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Literature summary extracted from

  • Willis, L.M.; Zhang, R.; Reid, A.; Withers, S.G.; Wakarchuk, W.W.
    Mechanistic investigation of the endo-alpha-N-acetylgalactosaminidase from Streptococcus pneumoniae R6 (2009), Biochemistry, 48, 10334-10341.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.97 expression of wild-type and mutant enzymes in Escherichia coli strain AD202 Streptococcus pneumoniae R6

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.97 D658A site-directed mutagenesis, the substrate specificity and activity is altered compared to the wild-type enzyme Streptococcus pneumoniae R6
3.2.1.97 D764A site-directed mutagenesis, the substrate specificity and activity is altered compared to the wild-type enzyme Streptococcus pneumoniae R6
3.2.1.97 D764F site-directed mutagenesis, the substrate specificity and activity is altered compared to the wild-type enzyme Streptococcus pneumoniae R6
3.2.1.97 E796A site-directed mutagenesis, the substrate specificity and activity is altered compared to the wild-type enzyme Streptococcus pneumoniae R6
3.2.1.97 E796Q site-directed mutagenesis, the substrate specificity and activity is altered compared to the wild-type enzyme Streptococcus pneumoniae R6
3.2.1.97 additional information construction of a DELTA1-39 deletion and DELTA1-39,DELTA1568-1767 deletion mutants Streptococcus pneumoniae R6

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.97 additional information
-
 additional information kinetic analysis, overview Streptococcus pneumoniae R6
3.2.1.97 0.0012
-
 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, mutant E796Q Streptococcus pneumoniae R6
3.2.1.97 0.0013
-
 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, mutant E796A Streptococcus pneumoniae R6
3.2.1.97 0.021
-
 4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, mutant E796A Streptococcus pneumoniae R6
3.2.1.97 0.033
-
 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, mutant D764A Streptococcus pneumoniae R6
3.2.1.97 0.034
-
 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, DELTA1-39 deletion mutant Streptococcus pneumoniae R6
3.2.1.97 0.048
-
 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, DELTA1-39,DELTA1568-1767 deletion mutant Streptococcus pneumoniae R6
3.2.1.97 0.062
-
 4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, DELTA1-39 deletion mutant Streptococcus pneumoniae R6
3.2.1.97 0.212
-
 4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, mutant E796Q Streptococcus pneumoniae R6
3.2.1.97 2.1
-
 2,4-dinitrophenyl-GalNAc-alpha-R pH 6.5, 37°C, mutant E796A Streptococcus pneumoniae R6
3.2.1.97 2.4
-
 2,4-dinitrophenyl-GalNAc-alpha-R pH 6.5, 37°C, DELTA1-39 deletion mutant Streptococcus pneumoniae R6

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.2.1.97 NaCl effects on kinetics, overview Streptococcus pneumoniae R6
3.2.1.97 NaN3 effects on kinetics, overview Streptococcus pneumoniae R6
3.2.1.97 Sodium formate effects on kinetics, overview Streptococcus pneumoniae R6

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.2.1.97 additional information Streptococcus pneumoniae R6 SpGH101 specifically removes an O-linked disaccharide Gal-beta-1,3-GalNAc-alpha from glycoproteins ?
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.97 Streptococcus pneumoniae R6 Q8DR60
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.97 recombinant wild-type and mutant enzymes from Escherichia coli strain AD202 by anion exchange chromatography, ultrafiltration, gel filtration, and hydrophobic interaction chromatography Streptococcus pneumoniae R6

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.97 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc + H2O
-
 Streptococcus pneumoniae R6 2,4-dinitrophenol + beta-D-Gal-(1->3)-alpha-D-GalNAc
-
 ?
3.2.1.97 2,4-dinitrophenyl-GalNAc-alpha-R + H2O
-
 Streptococcus pneumoniae R6 2,4-dinitrophenol + GalNAc-alpha-R
-
 ?
3.2.1.97 4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc + H2O
-
 Streptococcus pneumoniae R6 4-nitrophenol + beta-D-Gal-(1->3)-alpha-D-GalNAc
-
 ?
3.2.1.97 additional information SpGH101 specifically removes an O-linked disaccharide Gal-beta-1,3-GalNAc-alpha from glycoproteins Streptococcus pneumoniae R6 ?
-
 ?
3.2.1.97 additional information the enzyme contains several putative carbohydrate binding modules, structure-function relationship, D764 and E796 are the nucleophile and general acid-base residues, respectively, overview Streptococcus pneumoniae R6 ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.97 More analysis of the three-dimensional structure of the recombinant protein, the enzyme contains several putative carbohydrate binding modules, structure-function relationship, overview Streptococcus pneumoniae R6

Synonyms

EC Number Synonyms Comment Organism
3.2.1.97 endo-alpha-N-acetylgalactosaminidase
-
 Streptococcus pneumoniae R6
3.2.1.97 More the enzyme belongs to the glycoside hydrolase family 101, GH101 Streptococcus pneumoniae R6
3.2.1.97 SpGH101
-
 Streptococcus pneumoniae R6

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.2.1.97 37
-
 assay at Streptococcus pneumoniae R6

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.97 0.76
-
 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, mutant D764A Streptococcus pneumoniae R6
3.2.1.97 1
-
 4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, mutant E796A Streptococcus pneumoniae R6
3.2.1.97 1.6
-
 2,4-dinitrophenyl-GalNAc-alpha-R pH 6.5, 37°C, DELTA1-39 deletion mutant Streptococcus pneumoniae R6
3.2.1.97 2.1
-
 4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, mutant E796Q Streptococcus pneumoniae R6
3.2.1.97 5
-
 2,4-dinitrophenyl-GalNAc-alpha-R pH 6.5, 37°C, mutant E796A Streptococcus pneumoniae R6
3.2.1.97 14
-
 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, mutant E796Q Streptococcus pneumoniae R6
3.2.1.97 18
-
 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, mutant E796A Streptococcus pneumoniae R6
3.2.1.97 541
-
 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, DELTA1-39 deletion mutant Streptococcus pneumoniae R6
3.2.1.97 579
-
 4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, DELTA1-39 deletion mutant Streptococcus pneumoniae R6
3.2.1.97 748
-
 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, DELTA1-39,DELTA1568-1767 deletion mutant Streptococcus pneumoniae R6

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.2.1.97 additional information
-
 bell-shaped dependence of kcat/Km upon pH Streptococcus pneumoniae R6
3.2.1.97 6.5
-
 assay at Streptococcus pneumoniae R6

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.97 0.0006
-
 2,4-dinitrophenyl-GalNAc-alpha-R pH 6.5, 37°C, DELTA1-39 deletion mutant Streptococcus pneumoniae R6
3.2.1.97 0.002
-
 2,4-dinitrophenyl-GalNAc-alpha-R pH 6.5, 37°C, mutant E796A Streptococcus pneumoniae R6
3.2.1.97 0.0099
-
 4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, mutant E796Q Streptococcus pneumoniae R6
3.2.1.97 0.023
-
 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, mutant D764A Streptococcus pneumoniae R6
3.2.1.97 0.048
-
 4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, mutant E796A Streptococcus pneumoniae R6
3.2.1.97 0.987
-
 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, mutant D658A Streptococcus pneumoniae R6
3.2.1.97 9.3
-
 4-nitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, DELTA1-39 deletion mutant Streptococcus pneumoniae R6
3.2.1.97 11.7
-
 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, mutant E796Q Streptococcus pneumoniae R6
3.2.1.97 13.8
-
 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, mutant E796A Streptococcus pneumoniae R6
3.2.1.97 15.6
-
 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, DELTA1-39,DELTA1568-1767 deletion mutant Streptococcus pneumoniae R6
3.2.1.97 15.9
-
 2,4-dinitrophenyl beta-D-Gal-(1->3)-alpha-D-GalNAc pH 6.5, 37°C, DELTA1-39 deletion mutant Streptococcus pneumoniae R6