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Literature summary for 3.2.1.4 extracted from

  • Voutilainen, S.; Murray, P.; Tuohy, M.; Koivula, A.
    Expression of Talaromyces emersonii cellobiohydrolase Cel7A in Saccharomyces cerevisiae and rational mutagenesis to improve its thermostability and activity (2010), Protein Eng. Des. Sel., 23, 69-79 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Saccharomyces cerevisiae Rasamsonia emersonii

Protein Variants

Protein Variants Comment Organism
G266C/D320C introduction of an additional disulfide bridge, decrease in activity Rasamsonia emersonii
G4C/A72C introduction of an additional disulfide bridge, improves thermostability Rasamsonia emersonii
G4C/A72C/N54C/P191C/T243C/A375C introduction of 3 additional disulfide bridges, improves thermostability Rasamsonia emersonii
N54C/P191C introduction of an additional disulfide bridge, improves thermostability Rasamsonia emersonii
Q190C/I200C introduction of an additional disulfide bridge, decrease in activity Rasamsonia emersonii
T243C/A375C introduction of an additional disulfide bridge, improves thermostability Rasamsonia emersonii

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.21
-
4-methylumbelliferyl beta-D-lactoside mutant N54C/P191C, pH 5.0, 22°C Rasamsonia emersonii
0.23
-
4-methylumbelliferyl beta-D-lactoside mutant G4C/A72C/N54C/P191C/T243C/A375C, pH 5.0, 22°C Rasamsonia emersonii
0.26
-
4-methylumbelliferyl beta-D-lactoside mutant G4C/A72C, pH 5.0, 22°C Rasamsonia emersonii
0.31
-
4-methylumbelliferyl beta-D-lactoside deglycosylated wild-type, pH 5.0, 22°C Rasamsonia emersonii
0.31
-
4-methylumbelliferyl beta-D-lactoside mutant T243C/A375C, pH 5.0, 22°C Rasamsonia emersonii
0.33
-
4-methylumbelliferyl beta-D-lactoside wild-type, pH 5.0, 22°C Rasamsonia emersonii

Organism

Organism UniProt Comment Textmining
Rasamsonia emersonii Q8TFL9
-
-

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein x * 51475, MALDI-TOF of recombinant protein, x * 47206, MALDI-TOF of recombinant deglycosylated protein Rasamsonia emersonii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-methylumbelliferyl beta-D-lactoside + H2O
-
Rasamsonia emersonii 4-methylumbelliferone + D-lactose
-
?
avicel + H2O
-
Rasamsonia emersonii ?
-
?

Subunits

Subunits Comment Organism
? x * 46842, calculated, x * 66000, SDS-PAGE, x * 51475, MALDI-TOF of recombinant protein, x * 47206, MALDI-TOF of recombinant deglycosylated protein Rasamsonia emersonii

Synonyms

Synonyms Comment Organism
Cel7A
-
Rasamsonia emersonii

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
74.5
-
melting temperature, deglycosylated wild-type Rasamsonia emersonii
75
-
melting temperature, wild-type Rasamsonia emersonii
78.5
-
melting temperature, mutant N54C/P191C Rasamsonia emersonii
79
-
melting temperature, mutant T243C/A375C Rasamsonia emersonii
80
-
melting temperature, mutant G4C/A72C Rasamsonia emersonii
84
-
melting temperature, mutant G4C/A72C/N54C/P191C/T243C/A375C Rasamsonia emersonii

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.46
-
4-methylumbelliferyl beta-D-lactoside mutant G4C/A72C/N54C/P191C/T243C/A375C, pH 5.0, 22°C Rasamsonia emersonii
0.47
-
4-methylumbelliferyl beta-D-lactoside mutant T243C/A375C, pH 5.0, 22°C Rasamsonia emersonii
0.5
-
4-methylumbelliferyl beta-D-lactoside mutant G4C/A72C, pH 5.0, 22°C Rasamsonia emersonii
0.52
-
4-methylumbelliferyl beta-D-lactoside wild-type, pH 5.0, 22°C Rasamsonia emersonii
0.55
-
4-methylumbelliferyl beta-D-lactoside deglycosylated wild-type, pH 5.0, 22°C Rasamsonia emersonii
0.6
-
4-methylumbelliferyl beta-D-lactoside mutant N54C/P191C, pH 5.0, 22°C Rasamsonia emersonii

pI Value

Organism Comment pI Value Maximum pI Value
Rasamsonia emersonii isoelectric focusing
-
4
Rasamsonia emersonii calculated
-
4.1

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.4
-
4-methylumbelliferyl beta-D-lactoside mutant T243C/A375C, pH 5.0, 22°C Rasamsonia emersonii
1.6
-
4-methylumbelliferyl beta-D-lactoside wild-type, pH 5.0, 22°C Rasamsonia emersonii
1.8
-
4-methylumbelliferyl beta-D-lactoside deglycosylated wild-type, pH 5.0, 22°C Rasamsonia emersonii
1.9
-
4-methylumbelliferyl beta-D-lactoside mutant G4C/A72C, pH 5.0, 22°C Rasamsonia emersonii
1.9
-
4-methylumbelliferyl beta-D-lactoside mutant G4C/A72C/N54C/P191C/T243C/A375C, pH 5.0, 22°C Rasamsonia emersonii
2.8
-
4-methylumbelliferyl beta-D-lactoside mutant N54C/P191C, pH 5.0, 22°C Rasamsonia emersonii