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Literature summary extracted from

  • Fan, S.; Huang, W.; Wang, L.
    Remarkable transglycosylation activity of glycosynthase mutants of Endo-D, an endo-beta-N-acetylglucosaminidase from Streptococcus pneumoniae (2012), J. Biol. Chem., 287, 11272-11281.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.2.1.96 expressed in Escherichia coli BL21(DE3) cells Streptococcus pneumoniae

Protein Variants

EC Number Protein Variants Comment Organism
3.2.1.96 H371W the mutant shows increased hydrolytic activity compared to the wild type Streptococcus pneumoniae
3.2.1.96 N322A the mutant demonstrates no product hydrolysis activity Streptococcus pneumoniae
3.2.1.96 N322A the mutant demonstrates remarkable transglycosylation activity with no product hydrolysis activity, having much higher catalytic efficiency for glycosylating the nonfucosylated GlcNAc acceptor Streptococcus pneumoniae
3.2.1.96 N322Q the mutant demonstrates remarkable transglycosylation activity with only marginal hydrolysis activity, having much higher catalytic efficiency for glycosylating the nonfucosylated GlcNAc acceptor Streptococcus pneumoniae
3.2.1.96 Y360F the mutant shows slightly reduced hydrolytic activity compared to the wild type Streptococcus pneumoniae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.2.1.96 0.5
-
 Man3GlcNAc-oxazoline mutant enzyme N322Q, with Fmoc-Asn(Fucalpha1,6GlcNAc)-OH as acceptor, at pH 7.5 and 30°C Streptococcus pneumoniae
3.2.1.96 0.67
-
 Man3GlcNAc-oxazoline mutant enzyme N322A, with Fmoc-Asn(Fucalpha1,6GlcNAc)-OH as acceptor, at pH 7.5 and 30°C Streptococcus pneumoniae
3.2.1.96 0.73
-
 Fmoc-Asn(GlcNAc)-OH mutant enzyme N322Q, at pH 7.5 and 30°C Streptococcus pneumoniae
3.2.1.96 5.22
-
 Fmoc-Asn(GlcNAc)-OH mutant enzyme N322A, at pH 7.5 and 30°C Streptococcus pneumoniae
3.2.1.96 8.51
-
 Fmoc-Asn(Fucalpha(1->6)GlcNAc)-OH mutant enzyme N322A, at pH 7.5 and 30°C Streptococcus pneumoniae
3.2.1.96 18.51
-
 Fmoc-Asn(Fucalpha(1->6)GlcNAc)-OH mutant enzyme N322Q, at pH 7.5 and 30°C Streptococcus pneumoniae

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.2.1.96 100000
-
 x * 100000, SDS-PAGE Streptococcus pneumoniae
3.2.1.96 102000
-
 x * 102000, calculated from amino acid sequence Streptococcus pneumoniae

Organism

EC Number Organism UniProt Comment Textmining
3.2.1.96 Streptococcus pneumoniae Q93HW0
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.2.1.96 Ni2+-immobilized HisTrap column chromatography Streptococcus pneumoniae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.2.1.96 Man3GlcNAc-oxazoline + Fmoc-Asn(Fucalpha(1->6)GlcNAc)-OH
-
 Streptococcus pneumoniae ?
-
 ?
3.2.1.96 Man3GlcNAc-oxazoline + Fmoc-Asn(GlcNAc)-OH
-
 Streptococcus pneumoniae ?
-
 ?
3.2.1.96 additional information the enzyme also possesses transglycosylation activity with sugar oxazoline as the donor substrate, but the transglycosylation yield is low due to enzymatic hydrolysis of the donor substrate and the product. The enzyme favors nonfucosylated GlcNAc acceptor for transglycosylation Streptococcus pneumoniae ?
-
 ?
3.2.1.96 N-glycosylated immunglobuline G + H2O the enzyme prefers core-fucosylated N-glycan for hydrolysis Streptococcus pneumoniae ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.2.1.96 ? x * 100000, SDS-PAGE Streptococcus pneumoniae
3.2.1.96 ? x * 102000, calculated from amino acid sequence Streptococcus pneumoniae

Synonyms

EC Number Synonyms Comment Organism
3.2.1.96 endo-beta-N-acetylglucosaminidase
-
 Streptococcus pneumoniae
3.2.1.96 Endo-D
-
 Streptococcus pneumoniae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.2.1.96 0.21
-
 Man3GlcNAc-oxazoline mutant enzyme N322A, with Fmoc-Asn(Fucalpha1,6GlcNAc)-OH as acceptor, at pH 7.5 and 30°C Streptococcus pneumoniae
3.2.1.96 0.24
-
 Fmoc-Asn(Fucalpha(1->6)GlcNAc)-OH mutant enzyme N322A, at pH 7.5 and 30°C Streptococcus pneumoniae
3.2.1.96 0.5
-
 Fmoc-Asn(GlcNAc)-OH mutant enzyme N322A, at pH 7.5 and 30°C Streptococcus pneumoniae
3.2.1.96 6.37
-
 Fmoc-Asn(GlcNAc)-OH mutant enzyme N322Q, at pH 7.5 and 30°C Streptococcus pneumoniae
3.2.1.96 10.17
-
 Man3GlcNAc-oxazoline mutant enzyme N322Q, with Fmoc-Asn(Fucalpha1,6GlcNAc)-OH as acceptor, at pH 7.5 and 30°C Streptococcus pneumoniae
3.2.1.96 14.32
-
 Fmoc-Asn(Fucalpha(1->6)GlcNAc)-OH mutant enzyme N322Q, at pH 7.5 and 30°C Streptococcus pneumoniae

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.2.1.96 0.0286
-
 Fmoc-Asn(Fucalpha(1->6)GlcNAc)-OH mutant enzyme N322A, at pH 7.5 and 30°C Streptococcus pneumoniae
3.2.1.96 0.096
-
 Fmoc-Asn(GlcNAc)-OH mutant enzyme N322A, at pH 7.5 and 30°C Streptococcus pneumoniae
3.2.1.96 0.31
-
 Man3GlcNAc-oxazoline mutant enzyme N322A, with Fmoc-Asn(Fucalpha1,6GlcNAc)-OH as acceptor, at pH 7.5 and 30°C Streptococcus pneumoniae
3.2.1.96 0.77
-
 Fmoc-Asn(Fucalpha(1->6)GlcNAc)-OH mutant enzyme N322Q, at pH 7.5 and 30°C Streptococcus pneumoniae
3.2.1.96 8.7
-
 Fmoc-Asn(GlcNAc)-OH mutant enzyme N322Q, at pH 7.5 and 30°C Streptococcus pneumoniae
3.2.1.96 20.4
-
 Man3GlcNAc-oxazoline mutant enzyme N322Q, with Fmoc-Asn(Fucalpha1,6GlcNAc)-OH as acceptor, at pH 7.5 and 30°C Streptococcus pneumoniae