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Literature summary for 3.2.1.91 extracted from

  • Boonvitthya, N.; Bozonnet, S.; Burapatana, V.; ODonohue, M.J.; Chulalaksananukul, W.
    Comparison of the heterologous expression of Trichoderma reesei endoglucanase II and cellobiohydrolase II in the yeasts Pichia pastoris and Yarrowia lipolytica (2013), Mol. Biotechnol., 54, 158-169.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene cbh2, recombinant expression of His-tagged enzyme in Pichia pastoris strain X-33 under the control of the AOX1 promoter, cloning and recombinant expression in Yarrowia lipolytica strain Po1d under the control of the POX2 or TEF promoters, and using either the native or preproLip2 secretion signals. The expression level of cellobiohydrolase from the TEF-preproLip2-cbhII cassette is higher in Yarrowia lipolytica than in Pichia pastoris Trichoderma reesei

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
65000
-
x * 60000-63000, recombinant enzyme from Yarrowia lipolytica , SDS-PAGE, x * 65000, recombinant His-tagged enzyme from Pichia pastoris , SDS-PAGE Trichoderma reesei

Organism

Organism UniProt Comment Textmining
Trichoderma reesei P07987 gene cbh2
-
Trichoderma reesei QM9414 P07987 gene cbh2
-

Posttranslational Modification

Posttranslational Modification Comment Organism
glycoprotein deglycosylation of the recombinant enzyme does not radically alter the specific activity Trichoderma reesei

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme 2.5fold from Yarrowia lipolytica by ultrafiltration and cation exchange chromatography, recombinant His-tagged enzyme 4.4fold from Pichia pastoris strain X-33 by metal affiity chromatography Trichoderma reesei

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.72
-
purified recombinant enzyme from Pichia pastoris, pH 5.0, 50°C Trichoderma reesei
2.4
-
purified recombinant enzyme from Yarrowia lipolytica, pH 5.0, 50°C Trichoderma reesei

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Avicel PH-101 + H2O
-
Trichoderma reesei ?
-
?
Avicel PH-101 + H2O
-
Trichoderma reesei QM9414 ?
-
?
phosphoric acid swollen cellulose + H2O
-
Trichoderma reesei ?
-
?
phosphoric acid swollen cellulose + H2O
-
Trichoderma reesei QM9414 ?
-
?

Subunits

Subunits Comment Organism
? x * 60000-63000, recombinant enzyme from Yarrowia lipolytica , SDS-PAGE, x * 65000, recombinant His-tagged enzyme from Pichia pastoris , SDS-PAGE Trichoderma reesei

Synonyms

Synonyms Comment Organism
CBHII
-
Trichoderma reesei
cellobiohydrolase II
-
Trichoderma reesei
PSCase
-
Trichoderma reesei

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
55
-
optimum activity around 55°C, followed by a sharp decline at temperatures above 60°C Trichoderma reesei

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40
-
purified recombinant enzymes, 30 min, good stability up to 40°C Trichoderma reesei
60
-
purified recombinant enzymes, loss of 50% of the initial activity after 30 min Trichoderma reesei

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
5.5 6
-
Trichoderma reesei

pH Stability

pH Stability pH Stability Maximum Comment Organism
5.5 6 purified recombinant enzymes, 100% stable in the range pH 5-6, undergoing rapid inactivation at alkaline pH and showing some instability below pH 5 Trichoderma reesei