Protein production by the filamentous fungus Trichoderma reesei

secretion of active antibody molecules

Eini Nyyssönen, Jan Demolder, Roland Contreras, Sirkka Keränen, Merja Penttilä

Research output: Contribution to journalArticleScientificpeer-review

Abstract

Trichoderma reesei is used by industry for production of plant material hydrolysing enzymes, especially cellulases. The fungus has excellent production and secretion capacity. The major cellulase cellobiohydrolase I (CBHI) represents half of the protein secreted and is encoded by a single copy gene. The strong cbh1 promoter and other promoters regulated in a different manner are available for protein production. The potential of the fungus in foreign protein production has been demonstrated by the expression of chymosin, interleukin-6, and laccase. Antibodies and their engineered forms have numerous applications. The capacity of Trichoderma to produce different forms of antibodies such as Fab molecules under the cbh1 promoter was analysed. When light chain was produced alone the secreted yields were very low but could be increased by introducing the heavy-Fd chain into the fungus. When the heavy-Fd chain was fused to the C-terminus of the CBHI core-linker region, production of secreted Fab's was increased about 50-fold. The amount of immunologically active CBHI – Fab molecules was about 150 mg/L in the medium in a fermenter cultivation. The released Fab molecules were authentic in their immunological properties demonstrating functional assembly of the light and heavy chains. The antibody part can be released from the CBHI fusion by an unidentified fungal protease or Kex2. The beneficial role of CBHI could be explained by more efficient transcription, ER entry or folding, or passage through the secretory pathway in general. Key words: heterologous expression, fusion protein, CBHI, Fab, single chain antibody.
Original languageEnglish
Pages (from-to)885-890
JournalCanadian Journal of Botany
Volume73
Issue numberS1
DOIs
Publication statusPublished - 1995
MoE publication typeA1 Journal article-refereed

Fingerprint

cellulose 1,4-beta-cellobiosidase
Trichoderma reesei
secretion
antibody
fungus
fungi
antibodies
protein
proteins
promoter regions
folding
chymosin
cellulases
laccase
fermenters
Trichoderma
enzyme
fold
endo-1,4-beta-glucanase
interleukin-6

Cite this

Nyyssönen, Eini ; Demolder, Jan ; Contreras, Roland ; Keränen, Sirkka ; Penttilä, Merja. / Protein production by the filamentous fungus Trichoderma reesei : secretion of active antibody molecules. In: Canadian Journal of Botany. 1995 ; Vol. 73, No. S1. pp. 885-890.
@article{070ff26f3a714347bd1984d3646bccf7,
title = "Protein production by the filamentous fungus Trichoderma reesei: secretion of active antibody molecules",
abstract = "Trichoderma reesei is used by industry for production of plant material hydrolysing enzymes, especially cellulases. The fungus has excellent production and secretion capacity. The major cellulase cellobiohydrolase I (CBHI) represents half of the protein secreted and is encoded by a single copy gene. The strong cbh1 promoter and other promoters regulated in a different manner are available for protein production. The potential of the fungus in foreign protein production has been demonstrated by the expression of chymosin, interleukin-6, and laccase. Antibodies and their engineered forms have numerous applications. The capacity of Trichoderma to produce different forms of antibodies such as Fab molecules under the cbh1 promoter was analysed. When light chain was produced alone the secreted yields were very low but could be increased by introducing the heavy-Fd chain into the fungus. When the heavy-Fd chain was fused to the C-terminus of the CBHI core-linker region, production of secreted Fab's was increased about 50-fold. The amount of immunologically active CBHI – Fab molecules was about 150 mg/L in the medium in a fermenter cultivation. The released Fab molecules were authentic in their immunological properties demonstrating functional assembly of the light and heavy chains. The antibody part can be released from the CBHI fusion by an unidentified fungal protease or Kex2. The beneficial role of CBHI could be explained by more efficient transcription, ER entry or folding, or passage through the secretory pathway in general. Key words: heterologous expression, fusion protein, CBHI, Fab, single chain antibody.",
author = "Eini Nyyss{\"o}nen and Jan Demolder and Roland Contreras and Sirkka Ker{\"a}nen and Merja Penttil{\"a}",
year = "1995",
doi = "10.1139/b95-335",
language = "English",
volume = "73",
pages = "885--890",
journal = "Botany",
issn = "1916-2790",
publisher = "National Research Council of Canada",
number = "S1",

}

Protein production by the filamentous fungus Trichoderma reesei : secretion of active antibody molecules. / Nyyssönen, Eini; Demolder, Jan; Contreras, Roland; Keränen, Sirkka; Penttilä, Merja.

In: Canadian Journal of Botany, Vol. 73, No. S1, 1995, p. 885-890.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Protein production by the filamentous fungus Trichoderma reesei

T2 - secretion of active antibody molecules

AU - Nyyssönen, Eini

AU - Demolder, Jan

AU - Contreras, Roland

AU - Keränen, Sirkka

AU - Penttilä, Merja

PY - 1995

Y1 - 1995

N2 - Trichoderma reesei is used by industry for production of plant material hydrolysing enzymes, especially cellulases. The fungus has excellent production and secretion capacity. The major cellulase cellobiohydrolase I (CBHI) represents half of the protein secreted and is encoded by a single copy gene. The strong cbh1 promoter and other promoters regulated in a different manner are available for protein production. The potential of the fungus in foreign protein production has been demonstrated by the expression of chymosin, interleukin-6, and laccase. Antibodies and their engineered forms have numerous applications. The capacity of Trichoderma to produce different forms of antibodies such as Fab molecules under the cbh1 promoter was analysed. When light chain was produced alone the secreted yields were very low but could be increased by introducing the heavy-Fd chain into the fungus. When the heavy-Fd chain was fused to the C-terminus of the CBHI core-linker region, production of secreted Fab's was increased about 50-fold. The amount of immunologically active CBHI – Fab molecules was about 150 mg/L in the medium in a fermenter cultivation. The released Fab molecules were authentic in their immunological properties demonstrating functional assembly of the light and heavy chains. The antibody part can be released from the CBHI fusion by an unidentified fungal protease or Kex2. The beneficial role of CBHI could be explained by more efficient transcription, ER entry or folding, or passage through the secretory pathway in general. Key words: heterologous expression, fusion protein, CBHI, Fab, single chain antibody.

AB - Trichoderma reesei is used by industry for production of plant material hydrolysing enzymes, especially cellulases. The fungus has excellent production and secretion capacity. The major cellulase cellobiohydrolase I (CBHI) represents half of the protein secreted and is encoded by a single copy gene. The strong cbh1 promoter and other promoters regulated in a different manner are available for protein production. The potential of the fungus in foreign protein production has been demonstrated by the expression of chymosin, interleukin-6, and laccase. Antibodies and their engineered forms have numerous applications. The capacity of Trichoderma to produce different forms of antibodies such as Fab molecules under the cbh1 promoter was analysed. When light chain was produced alone the secreted yields were very low but could be increased by introducing the heavy-Fd chain into the fungus. When the heavy-Fd chain was fused to the C-terminus of the CBHI core-linker region, production of secreted Fab's was increased about 50-fold. The amount of immunologically active CBHI – Fab molecules was about 150 mg/L in the medium in a fermenter cultivation. The released Fab molecules were authentic in their immunological properties demonstrating functional assembly of the light and heavy chains. The antibody part can be released from the CBHI fusion by an unidentified fungal protease or Kex2. The beneficial role of CBHI could be explained by more efficient transcription, ER entry or folding, or passage through the secretory pathway in general. Key words: heterologous expression, fusion protein, CBHI, Fab, single chain antibody.

U2 - 10.1139/b95-335

DO - 10.1139/b95-335

M3 - Article

VL - 73

SP - 885

EP - 890

JO - Botany

JF - Botany

SN - 1916-2790

IS - S1

ER -