Characterization of the activity profile in cellulases derived from recombinant strains of Trichoderma reesei

F. A. Zandona, P. D. Cancelier, Matti Siika-aho, L. P. Ramos

Research output: Chapter in Book/Report/Conference proceedingChapter or book articleProfessional

Abstract

Enzyme preparations from eight deletion strains ofTrichoderma reesei were characterized with regard to their activity profile against several model substrates, and subsequently compared to the complete cellulase system of the industrial preparation Celluclast 1.5L (Novozymes, Denmark). The recombinant enzymes (Röhm Enzyme Finland Oy, Finland) comprised of 4 single (A, CBH I-; B, CBH II-; C, EG I-; D, EG II-), 2 double- ( E, CBH I/II-, F, EG I/II-) and 2 trideletion (G, EG II-/CBH I/II-; H, CBH II-/EG I/II-) enzymes. Compared to Celluclast 1.5L, deletion of both genes encoding for EG I and EG II (enzyme F) resulted in very little endoglucanase activity in the enzyme preparation, whereas the absence of both CBH I and CBH II (enzyme E) reduced filter paper activity by more than 75%. In general, for saccharification experiments in which less than 2% of the substrate was hydrolyzed to soluble sugars, EG-rich preparations (enzyme E at 200 mg/g) caused a considerable decrease in the degree of depolymerization (DP) of substrates with high amorphous character (e.g., filter paper fibers), whereas cellulose DP was not affected when substrates having a higher supramolecular organization were used (e.g., Avicel, Sigmacell or cotton fibers). By contrast, when CBH-rich preparations (enzyme F, 200 mg/g) were used, greater cellulose saccharification yields were observed (14%), and changes in cellulose DP were primarily due to the progressive removal of cellobiose residues from the termini of cellulose molecules.
Original languageEnglish
Title of host publicationApplications of Enzymes to Lignocellulosics
EditorsShawn D. Mansfield, John N. Saddler
Place of PublicationWashington, DC
PublisherAmerican Chemical Society
Pages285-303
ISBN (Print)0-8412-3831-6
DOIs
Publication statusPublished - 2003
MoE publication typeD2 Article in professional manuals or guides or professional information systems or text book material
Event223rd ACS National Meeting - Orlando, United States
Duration: 7 Apr 200211 Apr 2002

Publication series

NameACS Symposium Series
PublisherACS
Volume855
ISSN (Print)0097-6156
ISSN (Electronic)1947-5918

Conference

Conference223rd ACS National Meeting
CountryUnited States
CityOrlando
Period7/04/0211/04/02

Fingerprint

Trichoderma reesei
cellulases
enzymes
depolymerization
saccharification
cellulose
Finland
cellulosic fibers
cellobiose
gene deletion
lint cotton
endo-1,4-beta-glucanase
Denmark
enzyme activity
sugars

Cite this

Zandona, F. A., Cancelier, P. D., Siika-aho, M., & Ramos, L. P. (2003). Characterization of the activity profile in cellulases derived from recombinant strains of Trichoderma reesei. In S. D. Mansfield, & J. N. Saddler (Eds.), Applications of Enzymes to Lignocellulosics (pp. 285-303). Washington, DC: American Chemical Society. ACS Symposium Series, Vol.. 855 https://doi.org/10.1021/bk-2003-0855.ch017
Zandona, F. A. ; Cancelier, P. D. ; Siika-aho, Matti ; Ramos, L. P. / Characterization of the activity profile in cellulases derived from recombinant strains of Trichoderma reesei. Applications of Enzymes to Lignocellulosics. editor / Shawn D. Mansfield ; John N. Saddler. Washington, DC : American Chemical Society, 2003. pp. 285-303 (ACS Symposium Series, Vol. 855).
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title = "Characterization of the activity profile in cellulases derived from recombinant strains of Trichoderma reesei",
abstract = "Enzyme preparations from eight deletion strains ofTrichoderma reesei were characterized with regard to their activity profile against several model substrates, and subsequently compared to the complete cellulase system of the industrial preparation Celluclast 1.5L (Novozymes, Denmark). The recombinant enzymes (R{\"o}hm Enzyme Finland Oy, Finland) comprised of 4 single (A, CBH I-; B, CBH II-; C, EG I-; D, EG II-), 2 double- ( E, CBH I/II-, F, EG I/II-) and 2 trideletion (G, EG II-/CBH I/II-; H, CBH II-/EG I/II-) enzymes. Compared to Celluclast 1.5L, deletion of both genes encoding for EG I and EG II (enzyme F) resulted in very little endoglucanase activity in the enzyme preparation, whereas the absence of both CBH I and CBH II (enzyme E) reduced filter paper activity by more than 75{\%}. In general, for saccharification experiments in which less than 2{\%} of the substrate was hydrolyzed to soluble sugars, EG-rich preparations (enzyme E at 200 mg/g) caused a considerable decrease in the degree of depolymerization (DP) of substrates with high amorphous character (e.g., filter paper fibers), whereas cellulose DP was not affected when substrates having a higher supramolecular organization were used (e.g., Avicel, Sigmacell or cotton fibers). By contrast, when CBH-rich preparations (enzyme F, 200 mg/g) were used, greater cellulose saccharification yields were observed (14{\%}), and changes in cellulose DP were primarily due to the progressive removal of cellobiose residues from the termini of cellulose molecules.",
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Zandona, FA, Cancelier, PD, Siika-aho, M & Ramos, LP 2003, Characterization of the activity profile in cellulases derived from recombinant strains of Trichoderma reesei. in SD Mansfield & JN Saddler (eds), Applications of Enzymes to Lignocellulosics. American Chemical Society, Washington, DC, ACS Symposium Series, vol. 855, pp. 285-303, 223rd ACS National Meeting, Orlando, United States, 7/04/02. https://doi.org/10.1021/bk-2003-0855.ch017

Characterization of the activity profile in cellulases derived from recombinant strains of Trichoderma reesei. / Zandona, F. A.; Cancelier, P. D.; Siika-aho, Matti; Ramos, L. P.

Applications of Enzymes to Lignocellulosics. ed. / Shawn D. Mansfield; John N. Saddler. Washington, DC : American Chemical Society, 2003. p. 285-303 (ACS Symposium Series, Vol. 855).

Research output: Chapter in Book/Report/Conference proceedingChapter or book articleProfessional

TY - CHAP

T1 - Characterization of the activity profile in cellulases derived from recombinant strains of Trichoderma reesei

AU - Zandona, F. A.

AU - Cancelier, P. D.

AU - Siika-aho, Matti

AU - Ramos, L. P.

PY - 2003

Y1 - 2003

N2 - Enzyme preparations from eight deletion strains ofTrichoderma reesei were characterized with regard to their activity profile against several model substrates, and subsequently compared to the complete cellulase system of the industrial preparation Celluclast 1.5L (Novozymes, Denmark). The recombinant enzymes (Röhm Enzyme Finland Oy, Finland) comprised of 4 single (A, CBH I-; B, CBH II-; C, EG I-; D, EG II-), 2 double- ( E, CBH I/II-, F, EG I/II-) and 2 trideletion (G, EG II-/CBH I/II-; H, CBH II-/EG I/II-) enzymes. Compared to Celluclast 1.5L, deletion of both genes encoding for EG I and EG II (enzyme F) resulted in very little endoglucanase activity in the enzyme preparation, whereas the absence of both CBH I and CBH II (enzyme E) reduced filter paper activity by more than 75%. In general, for saccharification experiments in which less than 2% of the substrate was hydrolyzed to soluble sugars, EG-rich preparations (enzyme E at 200 mg/g) caused a considerable decrease in the degree of depolymerization (DP) of substrates with high amorphous character (e.g., filter paper fibers), whereas cellulose DP was not affected when substrates having a higher supramolecular organization were used (e.g., Avicel, Sigmacell or cotton fibers). By contrast, when CBH-rich preparations (enzyme F, 200 mg/g) were used, greater cellulose saccharification yields were observed (14%), and changes in cellulose DP were primarily due to the progressive removal of cellobiose residues from the termini of cellulose molecules.

AB - Enzyme preparations from eight deletion strains ofTrichoderma reesei were characterized with regard to their activity profile against several model substrates, and subsequently compared to the complete cellulase system of the industrial preparation Celluclast 1.5L (Novozymes, Denmark). The recombinant enzymes (Röhm Enzyme Finland Oy, Finland) comprised of 4 single (A, CBH I-; B, CBH II-; C, EG I-; D, EG II-), 2 double- ( E, CBH I/II-, F, EG I/II-) and 2 trideletion (G, EG II-/CBH I/II-; H, CBH II-/EG I/II-) enzymes. Compared to Celluclast 1.5L, deletion of both genes encoding for EG I and EG II (enzyme F) resulted in very little endoglucanase activity in the enzyme preparation, whereas the absence of both CBH I and CBH II (enzyme E) reduced filter paper activity by more than 75%. In general, for saccharification experiments in which less than 2% of the substrate was hydrolyzed to soluble sugars, EG-rich preparations (enzyme E at 200 mg/g) caused a considerable decrease in the degree of depolymerization (DP) of substrates with high amorphous character (e.g., filter paper fibers), whereas cellulose DP was not affected when substrates having a higher supramolecular organization were used (e.g., Avicel, Sigmacell or cotton fibers). By contrast, when CBH-rich preparations (enzyme F, 200 mg/g) were used, greater cellulose saccharification yields were observed (14%), and changes in cellulose DP were primarily due to the progressive removal of cellobiose residues from the termini of cellulose molecules.

U2 - 10.1021/bk-2003-0855.ch017

DO - 10.1021/bk-2003-0855.ch017

M3 - Chapter or book article

SN - 0-8412-3831-6

T3 - ACS Symposium Series

SP - 285

EP - 303

BT - Applications of Enzymes to Lignocellulosics

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PB - American Chemical Society

CY - Washington, DC

ER -

Zandona FA, Cancelier PD, Siika-aho M, Ramos LP. Characterization of the activity profile in cellulases derived from recombinant strains of Trichoderma reesei. In Mansfield SD, Saddler JN, editors, Applications of Enzymes to Lignocellulosics. Washington, DC: American Chemical Society. 2003. p. 285-303. (ACS Symposium Series, Vol. 855). https://doi.org/10.1021/bk-2003-0855.ch017