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Literature summary for 3.2.1.176 extracted from

  • Voutilainen, S.P.; Boer, H.; Alapuranen, M.; Jaenis, J.; Vehmaanperae, J.; Koivula, A.
    Improving the thermostability and activity of Melanocarpus albomyces cellobiohydrolase Cel7B (2009), Appl. Microbiol. Biotechnol., 83, 261-272.
    View publication on PubMed
Search for more literature for 3.2.1.176

Cloned(Commentary)

Cloned (Comment) Organism
wild-type and mutants expressed in Saccharomyces cerevisiae strain INVSc1. Cel7B wild-type and a two-module version containing as a C-terminal fusion the linker and cellulose-binding module (CBM) of Trichoderma reesei Cel7A expressed in Trichoderma reesei strain A36 Melanocarpus albomyces

Protein Variants

Protein Variants Comment Organism
G4C/M70C disulfide bridge mutation located near the N-terminus, close to the entrance of the active site tunnel of Cel7B, which leads to improved thermostability. The unfolding temperature is increased by 2.5°C compared to that of the wild-type. The mutant has increased activity towards microcrystalline cellulose (avicel) at 75°C Melanocarpus albomyces
G4C/M70C/S290T thermostability-increasing mutation together with disulfide bridge mutation, the unfolding temperature is increased by 4°C compared to that of the wild-type, additive effect on thermostability. The mutant has increased activity towards microcrystalline cellulose (avicel) at 75°C Melanocarpus albomyces
S290T can hydrolyse crystalline cellulose at 70°C 2fold more effectively than the wild-type, the unfolding temperature is increased by 1.5°C compared to that of the wild-type Melanocarpus albomyces

Inhibitors

Inhibitors Comment Organism Structure
cellobiose
-
 Melanocarpus albomyces

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.23
-
 4-methylumbelliferyl-beta-D-lactoside Cel7B wild-type expressed in Trichoderma reesei, at 22°C, pH 6.0 Melanocarpus albomyces
0.28
-
 4-methylumbelliferyl-beta-D-lactoside Cel7B mutant S290T expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
0.28
-
 4-methylumbelliferyl-beta-D-lactoside Cel7B wild-type expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
0.3
-
 4-methylumbelliferyl-beta-D-lactoside Cel7B mutant G4C/M70C expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
0.3
-
 4-methylumbelliferyl-beta-D-lactoside Cel7B mutant S290T/G4C/M70C expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
0.62
-
 2-chloro-4-nitrophenyl-beta-D-lactoside Cel7B wild-type expressed in Trichoderma reesei, at 22°C, pH 6.0 Melanocarpus albomyces
0.75
-
 2-chloro-4-nitrophenyl-beta-D-lactoside Cel7B mutant S290T/G4C/M70C expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
0.8
-
 2-chloro-4-nitrophenyl-beta-D-lactoside Cel7B wild-type expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
1
-
 2-chloro-4-nitrophenyl-beta-D-lactoside Cel7B mutant G4C/M70C expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
1.16
-
 2-chloro-4-nitrophenyl-beta-D-lactoside Cel7B mutant S290T expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
50000
-
 x * 50000, SDS-PAGE Melanocarpus albomyces

Organism

Organism UniProt Comment Textmining
Melanocarpus albomyces
-
-
-

Purification (Commentary)

Purification (Comment) Organism
wild-type and mutants purified with one-step purification using gel filtration Melanocarpus albomyces

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-chloro-4-nitrophenyl-beta-D-lactoside + H2O
-
 Melanocarpus albomyces 2-chloro-4-nitrophenol + D-lactose
-
 ?
4-methylumbelliferyl-beta-D-lactoside + H2O
-
 Melanocarpus albomyces 4-methylumbelliferol + D-lactose
-
 ?
avicel + H2O
-
 Melanocarpus albomyces ?
-
 ?

Subunits

Subunits Comment Organism
? x * 50000, SDS-PAGE Melanocarpus albomyces

Synonyms

Synonyms Comment Organism
Cel7B
-
 Melanocarpus albomyces
cellobiohydrolase
-
 Melanocarpus albomyces

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
additional information
-
 the Trichoderma reesei produced fusion protein Cel7B-CBM has 2.5°C improved unfolding temperature when compared to the Trichoderma reesei produced catalytic module (Cel7B wild-type) Melanocarpus albomyces

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0153
-
 2-chloro-4-nitrophenyl-beta-D-lactoside Cel7B wild-type expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
0.0165
-
 2-chloro-4-nitrophenyl-beta-D-lactoside Cel7B mutant S290T/G4C/M70C expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
0.0167
-
 2-chloro-4-nitrophenyl-beta-D-lactoside Cel7B wild-type expressed in Trichoderma reesei, at 22°C, pH 6.0 Melanocarpus albomyces
0.01817
-
 2-chloro-4-nitrophenyl-beta-D-lactoside Cel7B mutant G4C/M70C expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
0.0205
-
 2-chloro-4-nitrophenyl-beta-D-lactoside Cel7B mutant S290T expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
0.083
-
 4-methylumbelliferyl-beta-D-lactoside Cel7B mutant G4C/M70C expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
0.1067
-
 4-methylumbelliferyl-beta-D-lactoside Cel7B wild-type expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
0.1083
-
 4-methylumbelliferyl-beta-D-lactoside Cel7B mutant S290T expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
0.12
-
 4-methylumbelliferyl-beta-D-lactoside Cel7B mutant S290T/G4C/M70C expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
0.143
-
 4-methylumbelliferyl-beta-D-lactoside Cel7B wild-type expressed in Trichoderma reesei, at 22°C, pH 6.0 Melanocarpus albomyces

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.0055
-
 cellobiose Cel7B mutant G4C/M70C expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
0.006
-
 cellobiose Cel7B wild-type expressed in Trichoderma reesei, at 22°C, pH 6.0 Melanocarpus albomyces
0.0063
-
 cellobiose Cel7B wild-type expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
0.0066
-
 cellobiose Cel7B mutant S290T expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces
0.019
-
 cellobiose Cel7B mutant S290T/G4C/M70C expressed in Saccharomyces cerevisiae, at 22°C, pH 6.0 Melanocarpus albomyces