Pilot scale production of a heterologous Trichoderma reesei cellulase by Saccharomyces cerevisiae

Beat Zurbriggen, Michael J. Bailey, Merja E. Penttilä, Kaisa Poutanen, Matti Linko

Research output: Contribution to journalArticleScientificpeer-review

55 Citations (Scopus)

Abstract

Cellobiohydrolase II of Trichoderma reesei was produced in laboratory and pilot scale using a transformant strain of Saccharomyces cerevisiae harbouring a multicopy expression plasmid. Different strategies were compared for concentration and partial purification of the enzyme produced in a 200 l pilot cultivation. After efficient separation of biomass and sub-cellular particulate matter, a combination of ultrafiltration and adsorbent treatment for removal of protein impurities was used to provide a concentrate for chromatographic purification. Effective purification of the CBH II protein was obtained by passing the concentrate through a column of DEAE Sepharose, on which almost all the yeast proteins were adsorbed. The purified enzyme reacted with antibodies prepared against T. reesei CBH II and catalyzed partial solubilization of crystalline cellulose to soluble sugars.

Original languageEnglish
Pages (from-to)267-278
Number of pages12
JournalJournal of Biotechnology
Volume13
Issue number4
DOIs
Publication statusPublished - 1 Jan 1990
MoE publication typeA1 Journal article-refereed

Fingerprint

Trichoderma
Cellulase
Yeast
Purification
Saccharomyces cerevisiae
Cellulose 1,4-beta-Cellobiosidase
Fungal Proteins
Particulate Matter
Ultrafiltration
Enzymes
Cellulose
Biomass
Sepharose
Proteins
Plasmids
Antibodies
Sugars
Adsorbents
Impurities
Crystalline materials

Keywords

  • Downstream processing
  • Pilot production
  • Saccharomyces cerevisiae
  • Trichoderma reesei cellobiohydrolase II

Cite this

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abstract = "Cellobiohydrolase II of Trichoderma reesei was produced in laboratory and pilot scale using a transformant strain of Saccharomyces cerevisiae harbouring a multicopy expression plasmid. Different strategies were compared for concentration and partial purification of the enzyme produced in a 200 l pilot cultivation. After efficient separation of biomass and sub-cellular particulate matter, a combination of ultrafiltration and adsorbent treatment for removal of protein impurities was used to provide a concentrate for chromatographic purification. Effective purification of the CBH II protein was obtained by passing the concentrate through a column of DEAE Sepharose, on which almost all the yeast proteins were adsorbed. The purified enzyme reacted with antibodies prepared against T. reesei CBH II and catalyzed partial solubilization of crystalline cellulose to soluble sugars.",
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Pilot scale production of a heterologous Trichoderma reesei cellulase by Saccharomyces cerevisiae. / Zurbriggen, Beat; Bailey, Michael J.; Penttilä, Merja E.; Poutanen, Kaisa; Linko, Matti.

In: Journal of Biotechnology, Vol. 13, No. 4, 01.01.1990, p. 267-278.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Pilot scale production of a heterologous Trichoderma reesei cellulase by Saccharomyces cerevisiae

AU - Zurbriggen, Beat

AU - Bailey, Michael J.

AU - Penttilä, Merja E.

AU - Poutanen, Kaisa

AU - Linko, Matti

PY - 1990/1/1

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AB - Cellobiohydrolase II of Trichoderma reesei was produced in laboratory and pilot scale using a transformant strain of Saccharomyces cerevisiae harbouring a multicopy expression plasmid. Different strategies were compared for concentration and partial purification of the enzyme produced in a 200 l pilot cultivation. After efficient separation of biomass and sub-cellular particulate matter, a combination of ultrafiltration and adsorbent treatment for removal of protein impurities was used to provide a concentrate for chromatographic purification. Effective purification of the CBH II protein was obtained by passing the concentrate through a column of DEAE Sepharose, on which almost all the yeast proteins were adsorbed. The purified enzyme reacted with antibodies prepared against T. reesei CBH II and catalyzed partial solubilization of crystalline cellulose to soluble sugars.

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