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Mycology 2017
September 25-26, 2017
conferenceseries
.com
September 25-26, 2017 Chicago, USA
2
nd
International Conference on
Mycology & Mushrooms
Volume 3, Issue 2 (Suppl)
Med Mycol Open Access
ISSN: 2471-8521
Next generation biotherapeutic production system: The filamentous fungus
Trichoderma reesei
Christopher P Landowski
1
, Anne Huuskonen
1
, Ramon Wahl
2
, Ann Westerholm-Parvinen
1
, Benjamin Sommer
2
, Merja Penttilä
1
, Jari Natunen
3
, Christian
Ostermeier
2
, Bernhard Helk
2
, Juhani Saarinen
3
and
Markku Saloheimo
1
1
VTT Technical Research Centre of Finland, Finland
2
Novartis Pharma AG, Switzerland
3
Glykos, Finland
T
he filamentous fungus
Trichoderma reesei
is an important production organism used by industrial enzyme companies world-
wide. It is a low cost production system that secretes its native enzymes at levels exceeding 100 g/L of culture medium. Several
T. reesei
produced enzymes have obtained the generally recognized safe status by the Food and Drug Administration.
T. reesei
has
tremendous prospects to be a cost efficient and high yield system for producing therapeutic proteins. We have adapted the fungus
to become more suitable for bio-therapeutic production by reducing secreted protease activity and altering glycosylation pathways
needed for adding mammalian glycoforms. Expression strains for monoclonal antibodies, Fab antibody fragments, interferon alpha-
2b, insulin-like growth factor 1, and fibroblast growth factor 21 were constructed, cultivated in bioreactors, and expression levels were
measured from the culture medium. After deleting 13 of the most critical protease genes, the general secreted protease activity was
reduced over 30-fold. Monoclonal antibodies could be produced up to 7.6 g/L, Fab antibody fragments up to 8.2 g/L, interferon alpha-
2b at 7.9 g/L, and insulin-like growth factor fusion protein at 8 g/L. With protease inhibitor treatment, interferon alpha-2b could be
produced at over 10 g/L, insulin-like growth factor fusion protein at 19 g/L, and full length fibroblast growth factor 21 at 200 mg/L in
addition to a shorter form at 3.5 g/L. Human glycoforms such as G0 and FG0 were produced on monoclonal antibodies. Expression
levels and product quality improved dramatically after multiple protease deletions and optimization of culture conditions. While the
production levels achieved are already relatively high, the strains could be developed further to reach the 100 g/L potential of the
organism. This study demonstrates the excellent prospects of
T. reesei
as a host for therapeutic protein production.
christopher.landowski@vtt.fiMed Mycol Open Access, 3:2, 2017
DOI: 10.21767/2471-8521-C1-003