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Literature summary for 3.2.1.52 extracted from

  • Choi, K.H.; Seo, J.Y.; Park, K.M.; Park, C.S.; Cha, J.
    Characterization of glycosyl hydrolase family 3 beta-N-acetylglucosaminidases from Thermotoga maritima and Thermotoga neapolitana (2009), J. Biosci. Bioeng., 108, 455-459.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene cbsA, expression of the His-tagged enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha Thermotoga neapolitana
gene nagA, DNA and amino acid sequence determination and analysis, expression of the His-tagged enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH5alpha Thermotoga maritima MSB8

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.28
-
4-nitrophenyl N-acetyl-beta-D-glucosamine pH 7.5, 70°C, recombinant enzyme Thermotoga maritima MSB8
0.29
-
4-nitrophenyl tri-N-acetyl-beta-D-chitotriose pH 7.5, 70°C, recombinant enzyme Thermotoga maritima MSB8
0.3
-
4-nitrophenyl tri-N-acetyl-beta-D-chitotriose pH 7.5, 70°C, recombinant enzyme Thermotoga neapolitana
0.36
-
4-nitrophenyl N-acetyl-beta-D-glucosamine pH 7.5, 70°C, recombinant enzyme Thermotoga neapolitana
0.37
-
4-nitrophenyl di-N-acetyl-beta-D-chitobiose pH 7.5, 70°C, recombinant enzyme Thermotoga maritima MSB8
0.44
-
4-nitrophenyl di-N-acetyl-beta-D-chitobiose pH 7.5, 70°C, recombinant enzyme Thermotoga neapolitana

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
54000
-
2 * 54000, recombinant enzyme, SDS-PAGE Thermotoga neapolitana
54000
-
2 * 54000, recombinant enzyme, SDS-PAGE Thermotoga maritima MSB8

Organism

Organism UniProt Comment Textmining
Thermotoga maritima MSB8 Q9WZR6 gene nagA
-
Thermotoga neapolitana
-
gene cbsA
-
Thermotoga neapolitana KCCM 41025
-
gene cbsA
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli by metal affinity chromatography and gel filtration Thermotoga neapolitana
recombinant His-tagged enzyme from Escherichia coli by metal affinity chromatography and gel filtration Thermotoga maritima MSB8

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl di-N-acetyl-beta-D-chitobiose + H2O NagA initially hydrolyzes 4-nitrophenyl substrates to give GlcNAc Thermotoga neapolitana 4-nitrophenol + N-acetylglucosamine
-
?
4-nitrophenyl di-N-acetyl-beta-D-chitobiose + H2O NagA initially hydrolyzes 4-nitrophenyl substrates to give GlcNAc Thermotoga maritima MSB8 4-nitrophenol + N-acetylglucosamine
-
?
4-nitrophenyl di-N-acetyl-beta-D-chitobiose + H2O NagA initially hydrolyzes 4-nitrophenyl substrates to give GlcNAc Thermotoga neapolitana KCCM 41025 4-nitrophenol + N-acetylglucosamine
-
?
4-nitrophenyl N-acetyl-beta-D-glucosamine + H2O CbsA initially hydrolyzes 4-nitrophenyl substrates to give GlcNAc Thermotoga neapolitana 4-nitrophenol + N-acetylglucosamine
-
?
4-nitrophenyl N-acetyl-beta-D-glucosamine + H2O NagA initially hydrolyzes 4-nitrophenyl substrates to give GlcNAc Thermotoga maritima MSB8 4-nitrophenol + N-acetylglucosamine
-
?
4-nitrophenyl N-acetyl-beta-D-glucosamine + H2O CbsA initially hydrolyzes 4-nitrophenyl substrates to give GlcNAc Thermotoga neapolitana KCCM 41025 4-nitrophenol + N-acetylglucosamine
-
?
4-nitrophenyl tri-N-acetyl-beta-D-chitotriose + H2O NagA initially hydrolyzes 4-nitrophenyl substrates to give GlcNAc Thermotoga neapolitana 4-nitrophenol + N-acetylglucosamine
-
?
4-nitrophenyl tri-N-acetyl-beta-D-chitotriose + H2O NagA initially hydrolyzes 4-nitrophenyl substrates to give GlcNAc Thermotoga maritima MSB8 4-nitrophenol + N-acetylglucosamine
-
?
4-nitrophenyl tri-N-acetyl-beta-D-chitotriose + H2O NagA initially hydrolyzes 4-nitrophenyl substrates to give GlcNAc Thermotoga neapolitana KCCM 41025 4-nitrophenol + N-acetylglucosamine
-
?
di-N-acetyl-beta-D-chitobiose + H2O
-
Thermotoga neapolitana N-acetylglucosamine
-
?
di-N-acetyl-beta-D-chitobiose + H2O
-
Thermotoga maritima MSB8 N-acetylglucosamine
-
?
di-N-acetyl-beta-D-chitobiose + H2O
-
Thermotoga neapolitana KCCM 41025 N-acetylglucosamine
-
?
additional information the enzyme shows exo-activity with chitin oligosaccharides from the non-reducing ends, no activity with 4-nitrophenyl-beta-D-galactose and 4-nitrophenyl N-acetyl-beta-D-galactosamine, or highly polymerized chitin Thermotoga neapolitana ?
-
?
additional information the enzyme shows exo-activity with chitin oligosaccharides from the non-reducing ends, no activity with 4-nitrophenyl-beta-D-galactose and 4-nitrophenyl N-acetyl-beta-D-galactosamine, or highly polymerized chitin Thermotoga maritima MSB8 ?
-
?
additional information the enzyme shows exo-activity with chitin oligosaccharides from the non-reducing ends, no activity with 4-nitrophenyl-beta-D-galactose and 4-nitrophenyl N-acetyl-beta-D-galactosamine, or highly polymerized chitin Thermotoga neapolitana KCCM 41025 ?
-
?
tri-N-acetyl-beta-D-chitotriose + H2O
-
Thermotoga neapolitana N-acetylglucosamine
-
?
tri-N-acetyl-beta-D-chitotriose + H2O
-
Thermotoga maritima MSB8 N-acetylglucosamine
-
?

Subunits

Subunits Comment Organism
dimer 2 * 54000, recombinant enzyme, SDS-PAGE Thermotoga neapolitana
dimer 2 * 54000, recombinant enzyme, SDS-PAGE Thermotoga maritima MSB8

Synonyms

Synonyms Comment Organism
beta-N-acetylglucosaminidase
-
Thermotoga neapolitana
beta-N-acetylglucosaminidase
-
Thermotoga maritima MSB8
CbsA
-
Thermotoga neapolitana
More the enzyme belongs to the glycosyl hydrolase family 3, GH3 Thermotoga neapolitana
More the enzyme belongs to the glycosyl hydrolase family 3, GH3 Thermotoga maritima MSB8
NagA
-
Thermotoga maritima MSB8

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
65
-
-
Thermotoga maritima MSB8
75
-
-
Thermotoga neapolitana

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
50 95 activity profile, over 65% of maximal activity at 95°C, overview Thermotoga neapolitana
50 95 activity profile, over 65% of maximal activity at 95°C, overview Thermotoga maritima MSB8

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
146.7
-
4-nitrophenyl tri-N-acetyl-beta-D-chitotriose pH 7.5, 70°C, recombinant enzyme Thermotoga maritima MSB8
147.2
-
4-nitrophenyl tri-N-acetyl-beta-D-chitotriose pH 7.5, 70°C, recombinant enzyme Thermotoga neapolitana
199
-
4-nitrophenyl di-N-acetyl-beta-D-chitobiose pH 7.5, 70°C, recombinant enzyme Thermotoga maritima MSB8
220.8
-
4-nitrophenyl di-N-acetyl-beta-D-chitobiose pH 7.5, 70°C, recombinant enzyme Thermotoga neapolitana
246.2
-
4-nitrophenyl N-acetyl-beta-D-glucosamine pH 7.5, 70°C, recombinant enzyme Thermotoga maritima MSB8
273.4
-
4-nitrophenyl N-acetyl-beta-D-glucosamine pH 7.5, 70°C, recombinant enzyme Thermotoga neapolitana

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7 8
-
Thermotoga neapolitana
7 8
-
Thermotoga maritima MSB8

pH Range

pH Minimum pH Maximum Comment Organism
4.5 10 over 80% of maximal activity within the range of pH 6.5-8.5, rapid decline over pH 9.0, activity profile, overview Thermotoga neapolitana
4.5 10 over 80% of maximal activity within the range of pH 6.5-8.5, rapid decline over pH 9.0, activity profile, overview Thermotoga maritima MSB8

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
490.7
-
4-nitrophenyl tri-N-acetyl-beta-D-chitotriose pH 7.5, 70°C, recombinant enzyme Thermotoga neapolitana
501.2
-
4-nitrophenyl di-N-acetyl-beta-D-chitobiose pH 7.5, 70°C, recombinant enzyme Thermotoga neapolitana
505.7
-
4-nitrophenyl tri-N-acetyl-beta-D-chitotriose pH 7.5, 70°C, recombinant enzyme Thermotoga maritima MSB8
538
-
4-nitrophenyl di-N-acetyl-beta-D-chitobiose pH 7.5, 70°C, recombinant enzyme Thermotoga maritima MSB8
759.4
-
4-nitrophenyl N-acetyl-beta-D-glucosamine pH 7.5, 70°C, recombinant enzyme Thermotoga neapolitana
879.3
-
4-nitrophenyl N-acetyl-beta-D-glucosamine pH 7.5, 70°C, recombinant enzyme Thermotoga maritima MSB8