Enabling low cost biopharmaceuticals: high level interferon alpha-2b production in Trichoderma reesei

Christopher P. Landowski, Eero Mustalahti, Ramon Wahl, Laurence Croute, Dhinakaran Sivasiddarthan, Ann Westerholm-Parvinen, Benjamin Sommer, Christian Ostermeier, Bernhard Helk, Juhani Saarinen, Markku Saloheimo

Research output: Contribution to journalArticleScientificpeer-review

9 Citations (Scopus)

Abstract

Background: The filamentous fungus Trichoderma reesei has tremendous capability to secrete over 100 g/L of proteins and therefore it would make an excellent host system for production of high levels of therapeutic proteins at low cost. We have developed T. reesei strains suitable for production of therapeutic proteins by reducing the secreted protease activity. Protease activity has been the major hindrance to achieving high production levels. We have constructed a series of interferon alpha-2b (IFNa-2b) production strains with 9 protease deletions to gain knowledge for further strain development. Results: We have identified two protease deletions that dramatically improved the production levels. Deletion of the subtilisin protease slp7 and the metalloprotease amp2 has enabled production levels of IFNa-2b up to 2.1 and 2.4 g/L, respectively. With addition of soybean trypsin protease inhibitor the level of production improved to 4.5 g/L, with an additional 1.8 g/L still bound to the secretion carrier protein. Conclusions: High levels of IFNa-2b were produced using T. reesei strains with reduced protease secretion. Further strain development can be done to improve the production system by reducing protease activity and improving carrier protein cleavage.
Original languageEnglish
Article number104
JournalMicrobial Cell Factories
Volume15
Issue number1
DOIs
Publication statusPublished - 2016
MoE publication typeA1 Journal article-refereed

Fingerprint

interferon alfa-2b
Interferons
Trichoderma
Peptide Hydrolases
Costs and Cost Analysis
Costs
Proteins
Carrier Proteins
Subtilisin
Trypsin Inhibitors
Metalloproteases
Protease Inhibitors
Soybeans
Fungi

Keywords

  • Trichoderma reesei
  • Filamentous fungi
  • Interferon
  • Therapeutic proteins
  • Proteases
  • Cytokine

Cite this

Landowski, Christopher P. ; Mustalahti, Eero ; Wahl, Ramon ; Croute, Laurence ; Sivasiddarthan, Dhinakaran ; Westerholm-Parvinen, Ann ; Sommer, Benjamin ; Ostermeier, Christian ; Helk, Bernhard ; Saarinen, Juhani ; Saloheimo, Markku. / Enabling low cost biopharmaceuticals: high level interferon alpha-2b production in Trichoderma reesei. In: Microbial Cell Factories. 2016 ; Vol. 15, No. 1.
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abstract = "Background: The filamentous fungus Trichoderma reesei has tremendous capability to secrete over 100 g/L of proteins and therefore it would make an excellent host system for production of high levels of therapeutic proteins at low cost. We have developed T. reesei strains suitable for production of therapeutic proteins by reducing the secreted protease activity. Protease activity has been the major hindrance to achieving high production levels. We have constructed a series of interferon alpha-2b (IFNa-2b) production strains with 9 protease deletions to gain knowledge for further strain development. Results: We have identified two protease deletions that dramatically improved the production levels. Deletion of the subtilisin protease slp7 and the metalloprotease amp2 has enabled production levels of IFNa-2b up to 2.1 and 2.4 g/L, respectively. With addition of soybean trypsin protease inhibitor the level of production improved to 4.5 g/L, with an additional 1.8 g/L still bound to the secretion carrier protein. Conclusions: High levels of IFNa-2b were produced using T. reesei strains with reduced protease secretion. Further strain development can be done to improve the production system by reducing protease activity and improving carrier protein cleavage.",
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Enabling low cost biopharmaceuticals: high level interferon alpha-2b production in Trichoderma reesei. / Landowski, Christopher P.; Mustalahti, Eero; Wahl, Ramon; Croute, Laurence; Sivasiddarthan, Dhinakaran; Westerholm-Parvinen, Ann; Sommer, Benjamin; Ostermeier, Christian; Helk, Bernhard; Saarinen, Juhani; Saloheimo, Markku.

In: Microbial Cell Factories, Vol. 15, No. 1, 104, 2016.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Enabling low cost biopharmaceuticals: high level interferon alpha-2b production in Trichoderma reesei

AU - Landowski, Christopher P.

AU - Mustalahti, Eero

AU - Wahl, Ramon

AU - Croute, Laurence

AU - Sivasiddarthan, Dhinakaran

AU - Westerholm-Parvinen, Ann

AU - Sommer, Benjamin

AU - Ostermeier, Christian

AU - Helk, Bernhard

AU - Saarinen, Juhani

AU - Saloheimo, Markku

PY - 2016

Y1 - 2016

N2 - Background: The filamentous fungus Trichoderma reesei has tremendous capability to secrete over 100 g/L of proteins and therefore it would make an excellent host system for production of high levels of therapeutic proteins at low cost. We have developed T. reesei strains suitable for production of therapeutic proteins by reducing the secreted protease activity. Protease activity has been the major hindrance to achieving high production levels. We have constructed a series of interferon alpha-2b (IFNa-2b) production strains with 9 protease deletions to gain knowledge for further strain development. Results: We have identified two protease deletions that dramatically improved the production levels. Deletion of the subtilisin protease slp7 and the metalloprotease amp2 has enabled production levels of IFNa-2b up to 2.1 and 2.4 g/L, respectively. With addition of soybean trypsin protease inhibitor the level of production improved to 4.5 g/L, with an additional 1.8 g/L still bound to the secretion carrier protein. Conclusions: High levels of IFNa-2b were produced using T. reesei strains with reduced protease secretion. Further strain development can be done to improve the production system by reducing protease activity and improving carrier protein cleavage.

AB - Background: The filamentous fungus Trichoderma reesei has tremendous capability to secrete over 100 g/L of proteins and therefore it would make an excellent host system for production of high levels of therapeutic proteins at low cost. We have developed T. reesei strains suitable for production of therapeutic proteins by reducing the secreted protease activity. Protease activity has been the major hindrance to achieving high production levels. We have constructed a series of interferon alpha-2b (IFNa-2b) production strains with 9 protease deletions to gain knowledge for further strain development. Results: We have identified two protease deletions that dramatically improved the production levels. Deletion of the subtilisin protease slp7 and the metalloprotease amp2 has enabled production levels of IFNa-2b up to 2.1 and 2.4 g/L, respectively. With addition of soybean trypsin protease inhibitor the level of production improved to 4.5 g/L, with an additional 1.8 g/L still bound to the secretion carrier protein. Conclusions: High levels of IFNa-2b were produced using T. reesei strains with reduced protease secretion. Further strain development can be done to improve the production system by reducing protease activity and improving carrier protein cleavage.

KW - Trichoderma reesei

KW - Filamentous fungi

KW - Interferon

KW - Therapeutic proteins

KW - Proteases

KW - Cytokine

U2 - 10.1186/s12934-016-0508-5

DO - 10.1186/s12934-016-0508-5

M3 - Article

VL - 15

JO - Microbial Cell Factories

JF - Microbial Cell Factories

SN - 1475-2859

IS - 1

M1 - 104

ER -