Abstract
Brazzein, a small, pH-stable, and heat-stable sweet protein, is a promising alternative to sucrose because of its extreme sweetness and stability. However, large-scale production through traditional extraction methods from plant sources is not economically feasible. To this end, the current 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, liquid chromatography high-content mass spectroscopy (LC-HCMS), Timegated™ Raman spectroscopy, dynamic light scattering (DLS), and nanoscale differential scanning fluorimetry (nanoDSF). The results confirmed that the correct folding and stability of brazzein attributed to its robust internal structure, demonstrating the potential of T. reesei as an efficient host for producing this sweet protein.
| Original language | English |
|---|---|
| Article number | 1688495 |
| Number of pages | 14 |
| Journal | Frontiers in Bioengineering and Biotechnology |
| Volume | 13 |
| DOIs | |
| Publication status | Published - 2025 |
| MoE publication type | A1 Journal article-refereed |
Funding
The authors declare that financial support was received for the research and/or publication of this article. The authors thank VTT for providing funding for the study.
Keywords
- brazzein
- fungal expression system
- Raman spectroscopy
- sweet protein
- Trichoderma