ORIGINAL RESEARCH article
Front. Bioeng. Biotechnol.
Sec. Industrial Biotechnology
Label-free Analytical Characterization of Brazzein Produced with the Filamentous Fungus Trichoderma reesei
Provisionally accepted
Dominik Mojzita1
Nina Aro1
Juha Kontturi1
Martin Kögler2Emilia Nordlund1
Waltteri Hosia1
Marco G. Casteleijn1*- 1VTT Technical Research Centre of Finland Ltd, Espoo, Finland
- 2VTT Technical research Centre of Finland Ltd., Oulu, Finland
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Brazzein, a small, pH-and heat-stable sweet protein, is a promising alternative to sucrose due to its extreme sweetness and stability. However, large-scale production through traditional extraction methods from plant sources is not economically feasible. This study explores the production of recombinant brazzein in the filamentous fungus Trichoderma reesei using a synthetic expression system (SES) and the native fungal cbh1 promoter. Both methods successfully expressed brazzein in 24-well liquid cultures, with the SES system achieving a production level of ~ 1.3 g/L in bioreactor cultures. The protein was purified and characterized using SDS-PAGE, LC-HCMS, TimeGatedTM Raman spectroscopy, dynamic light scattering (DLS), and nano differential scanning fluorimetry (nanoDSF). The results confirmed the correct folding and stability of brazzein, demonstrating the potential of T. reesei as an efficient host for producing this sweet protein.
Keywords: Brazzein, Raman spectroscopy, mass spectroscopy, Fungal expression system, Sweet protein
Received: 19 Aug 2025; Accepted: 07 Nov 2025.
Copyright: © 2025 Mojzita, Aro, Kontturi, Kögler, Nordlund, Hosia and Casteleijn. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Marco G. Casteleijn, marco.casteleijn@vtt.fi
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