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Literature summary for 3.2.1.31 extracted from

  • Salleh, H.; Müllegger, J.; Reid, S.; Chan, W.; Hwang, J.; Warren, R.; Withers, S.
    Cloning and characterization of Thermotoga maritima beta-glucuronidase (2006), Carbohydr. Res., 341, 49-59 .
    View publication on PubMed
Search for more literature for 3.2.1.31

Application

Application Comment Organism
synthesis the enzyme appears suitable for use in enzymatic oligosaccharide synthesis in either the transglycosylation mode or by use of glycosynthase and thioglycoligase approaches Thermotoga maritima

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Thermotoga maritima

Protein Variants

Protein Variants Comment Organism
E383A mutant E383A and wild-type enzyme show a similar melting temperatures of about 85°C Thermotoga maritima
E383Q mutant enzyme has a reduced thermostability, becoming unfolded at 72°C Thermotoga maritima
Q476A mutant is hydrolytically inactive Thermotoga maritima

Inhibitors

Inhibitors Comment Organism Structure
2-deoxy-2-fluoro-beta-D-glucosyluronic acid fluoride mechanism-based inactivator, D-saccharic acid 1,4-lactone protects the enzyme against inactivation Thermotoga maritima

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0046
-
 4-nitrophenyl beta-D-glucuronide 65°C, pH 6.5, mutant enzyme E383Q Thermotoga maritima
0.0065
-
 4-nitrophenyl beta-D-glucuronide 65°C, pH 6.5, mutant enzyme E383A Thermotoga maritima
0.083
-
 4-nitrophenyl beta-D-glucuronide 65°C, pH 6.5, wild-type enzyme Thermotoga maritima
0.15
-
 4-nitrophenyl beta-D-glucuronide 75°C, pH 6.5, wild-type enzyme Thermotoga maritima
3.06
-
 4-nitrophenyl beta-D-galactosiduronide 75°C, pH 6.5, wild-type enzyme Thermotoga maritima
145
-
 4-nitrophenyl beta-D-glucoside 75°C, pH 6.5, wild-type enzyme Thermotoga maritima

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
66000
-
 SDS-PAGE Thermotoga maritima

Organism

Organism UniProt Comment Textmining
Thermotoga maritima Q9X0F2
-
-
Thermotoga maritima DSM 3109 Q9X0F2
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Thermotoga maritima

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl beta-D-galactosiduronide + H2O the enzyme has a moderately broad specificity, hydrolysing a range of 4-nitrophenyl glycoside substrates, but has greatest activity on 4-nitrophenyl beta-D-glucuronide. The catalytic nucleophile is Glu476 within the sequence VTEFGAD Thermotoga maritima 4-nitrophenol + beta-D-galactosiduronic acid
-
 ?
4-nitrophenyl beta-D-galactosiduronide + H2O the enzyme has a moderately broad specificity, hydrolysing a range of 4-nitrophenyl glycoside substrates, but has greatest activity on 4-nitrophenyl beta-D-glucuronide. The catalytic nucleophile is Glu476 within the sequence VTEFGAD Thermotoga maritima DSM 3109 4-nitrophenol + beta-D-galactosiduronic acid
-
 ?
4-nitrophenyl beta-D-glucoside + H2O the enzyme has a moderately broad specificity, hydrolysing a range of 4-nitrophenyl glycoside substrates, but has greatest activity on 4-nitrophenyl beta-D-glucuronide. The catalytic nucleophile is Glu476 within the sequence VTEFGAD Thermotoga maritima 4-nitrophenol + D-glucose
-
 ?
4-nitrophenyl beta-D-glucoside + H2O the enzyme has a moderately broad specificity, hydrolysing a range of 4-nitrophenyl glycoside substrates, but has greatest activity on 4-nitrophenyl beta-D-glucuronide. The catalytic nucleophile is Glu476 within the sequence VTEFGAD Thermotoga maritima DSM 3109 4-nitrophenol + D-glucose
-
 ?
4-nitrophenyl beta-D-glucuronide + H2O the enzyme has a moderately broad specificity, hydrolysing a range of 4-nitrophenyl glycoside substrates, but has greatest activity on 4-nitrophenyl beta-D-glucuronide. The catalytic nucleophile is Glu476 within the sequence VTEFGAD Thermotoga maritima 4-nitrophenol + D-glucuronic acid
-
 ?
4-nitrophenyl beta-D-glucuronide + H2O the enzyme has a moderately broad specificity, hydrolysing a range of 4-nitrophenyl glycoside substrates, but has greatest activity on 4-nitrophenyl beta-D-glucuronide. The catalytic nucleophile is Glu476 within the sequence VTEFGAD Thermotoga maritima DSM 3109 4-nitrophenol + D-glucuronic acid
-
 ?

Subunits

Subunits Comment Organism
? x * 66000, SDS-PAGE Thermotoga maritima

Synonyms

Synonyms Comment Organism
TM1062
-
 Thermotoga maritima

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
62
-
 unfolding of the E383Q mutant can be observed within the first 8 min while the E383A mutant does not show any denaturation even after 1 h Thermotoga maritima
72
-
 mutant enzyme E383Q has a reduced thermostability, becoming unfolded at 72°C Thermotoga maritima
85
-
 half-life: 3 h. Mutant E383A and wild-type enzyme show a similar melting temperatures of about 85°C Thermotoga maritima

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.33
-
 4-nitrophenyl beta-D-glucuronide 65°C, pH 6.5, mutant enzyme E383A Thermotoga maritima
0.98
-
 4-nitrophenyl beta-D-glucuronide 65°C, pH 6.5, mutant enzyme E383Q Thermotoga maritima
9
-
 4-nitrophenyl beta-D-glucoside 75°C, pH 6.5, wild-type enzyme Thermotoga maritima
29
-
 4-nitrophenyl beta-D-glucuronide 65°C, pH 6.5, wild-type enzyme Thermotoga maritima
68
-
 4-nitrophenyl beta-D-glucuronide 75°C, pH 6.5, wild-type enzyme Thermotoga maritima
443
-
 4-nitrophenyl beta-D-galactosiduronide 75°C, pH 6.5, wild-type enzyme Thermotoga maritima

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
-
 Thermotoga maritima

pH Range

pH Minimum pH Maximum Comment Organism
4.5 7.5
-
 Thermotoga maritima

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.06
-
 4-nitrophenyl beta-D-glucoside 75°C, pH 6.5, wild-type enzyme Thermotoga maritima
51
-
 4-nitrophenyl beta-D-glucuronide 65°C, pH 6.5, mutant enzyme E383A Thermotoga maritima
145
-
 4-nitrophenyl beta-D-galactosiduronide 75°C, pH 6.5, wild-type enzyme Thermotoga maritima
213
-
 4-nitrophenyl beta-D-glucuronide 65°C, pH 6.5, mutant enzyme E383Q Thermotoga maritima
352
-
 4-nitrophenyl beta-D-glucuronide 65°C, pH 6.5, wild-type enzyme Thermotoga maritima
453
-
 4-nitrophenyl beta-D-glucuronide 75°C, pH 6.5, wild-type enzyme Thermotoga maritima