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Literature summary for 3.2.1.165 extracted from

  • Honda, Y.; Arai, S.; Suzuki, K.; Kitaoka, M.; Fushinobu, S.
    The crystal structure of an inverting glycoside hydrolase family 9 exo-beta-D-glucosaminidase and the design of glycosynthase (2016), Biochem. J., 473, 463-472 .
    View publication on PubMed
Search for more literature for 3.2.1.165

Cloned(Commentary)

Cloned (Comment) Organism
wild type and mutant enzymes are expressed in Escherichia coli BL21(DE3) cells Photobacterium profundum

Crystallization (Commentary)

Crystallization (Comment) Organism
sitting drop vapor diffusion method, using 10% (w/v) PEG 6000, 0.1 M HEPES/NaOH (pH 7.5) and 5% (w/v) 2-methyl-2,4-pentanediol Photobacterium profundum

Protein Variants

Protein Variants Comment Organism
D139A the mutation significantly decreases the hydrolytic activity compared to the wild type enzyme. The mutant enzyme exclusively exhibits glycosynthase activity using alpha-D-glucosaminyl fluoride and D-glucosamine as substrates Photobacterium profundum
D139E compared with the wild type, the hydrolytic activity of the mutant is significantly suppressed (less than 0.1%), and the fluoride release activity also decreases to less than 3% Photobacterium profundum
D139S the mutation significantly decreases the hydrolytic activity compared to the wild type enzyme Photobacterium profundum
D143A the mutation significantly decreases the hydrolytic activity compared to the wild type enzyme Photobacterium profundum
E555A the mutation significantly decreases the hydrolytic activity compared to the wild type enzyme Photobacterium profundum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.8
-
 chitobiose wild type enzyme, at pH 7.0 and 37°C Photobacterium profundum
1.8
-
 chitobiose mutant enzyme D139E, at pH 7.0 and 37°C Photobacterium profundum
3.5
-
 alpha-D-glucosaminyl fluoride mutant enzyme D139E, at pH 7.0 and 37°C Photobacterium profundum
7.7
-
 chitobiose mutant enzyme D139A, at pH 7.0 and 37°C Photobacterium profundum
15
-
 chitobiose mutant enzyme D139S, at pH 7.0 and 37°C Photobacterium profundum

Organism

Organism UniProt Comment Textmining
Photobacterium profundum Q6LUT2
-
-
Photobacterium profundum SS9 Q6LUT2
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA column chromatography Photobacterium profundum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
alpha-D-glucosaminyl fluoride + D-glucosamine
-
 Photobacterium profundum chitobiose + fluoride
-
 ?
alpha-D-glucosaminyl fluoride + D-glucosamine
-
 Photobacterium profundum SS9 chitobiose + fluoride
-
 ?
chitobiose + H2O
-
 Photobacterium profundum ?
-
 ?
chitobiose + H2O
-
 Photobacterium profundum SS9 ?
-
 ?

Synonyms

Synonyms Comment Organism
exo-beta-D-glucosaminidase
-
 Photobacterium profundum
GlcNase
-
 Photobacterium profundum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0047
-
 chitobiose mutant enzyme D139A, at pH 7.0 and 37°C Photobacterium profundum
0.071
-
 chitobiose mutant enzyme D139E, at pH 7.0 and 37°C Photobacterium profundum
0.073
-
 alpha-D-glucosaminyl fluoride mutant enzyme D139E, at pH 7.0 and 37°C Photobacterium profundum
0.54
-
 chitobiose mutant enzyme D139S, at pH 7.0 and 37°C Photobacterium profundum
87
-
 chitobiose wild type enzyme, at pH 7.0 and 37°C Photobacterium profundum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.00062
-
 chitobiose mutant enzyme D139A, at pH 7.0 and 37°C Photobacterium profundum
0.0019
-
 alpha-D-glucosaminyl fluoride mutant enzyme D139A, at pH 7.0 and 37°C Photobacterium profundum
0.0031
-
 alpha-D-glucosaminyl fluoride mutant enzyme D139S, at pH 7.0 and 37°C Photobacterium profundum
0.021
-
 alpha-D-glucosaminyl fluoride mutant enzyme D139E, at pH 7.0 and 37°C Photobacterium profundum
0.036
-
 chitobiose mutant enzyme D139S, at pH 7.0 and 37°C Photobacterium profundum
0.04
-
 chitobiose mutant enzyme D139E, at pH 7.0 and 37°C Photobacterium profundum
0.73
-
 alpha-D-glucosaminyl fluoride wild type enzyme, at pH 7.0 and 37°C Photobacterium profundum
49
-
 chitobiose wild type enzyme, at pH 7.0 and 37°C Photobacterium profundum