Paving the way for sustainable protein sources for the future by exploring the inactivation of endogenous protease from green biomass

Background: 
The increasing global demand for sustainable protein sources necessitates innovative approaches in food processing. Leafy biomass, such as alfalfa, has shown potential for local dietary protein in Denmark. However, previous studies have demonstrated that endogenous proteolysis occurs rapidly in screw-pressed alfalfa juice, compromising the protein yield.

Aim:
This project aims to purify one or more proteases from green biomass and explore non-thermal approaches for their inactivation (for example: pulsed electric field), starting with simple model systems and progressing to more complex matrices (e.g. reconstituted systems, alfalfa juice, etc). 

Methodology and Skills Development:
Throughout the project, you can develop expertise in:

• Protein purification techniques
• Enzyme kinetics
• Various enzyme inactivation methods 
• Mass spectrometry

Some parts of the project can be tailored to your personal interests, allowing you to develop particular skills that align with your career goals and aspirations.

Project Overview:
Results from the project will help us to developing effective inactivation strategies for processing of leafy biomasses. The student will also be aligned with a 10-15 people group working with alternative proteins and natural colorants, biorefinery, stability and application. Ad hoc supervision in the lab will be provided. At weekly meetings the student will present her/his work and discuss design of experiments, results and perspective of the work. The student will present her/his data at a midterm seminar.

References:

1. Koschuh, W., Povoden, G., Thang, V. H., Kromus, S., Kulbe, K. D., Novalin, S., & Krotscheck, C. (2004). Production of leaf protein concentrate from ryegrass (Lolium perenne x multiflorum) and alfalfa (Medicago sauva subsp. sativa). Comparison between heat coagulation/centrifugation and ultrafiltration. Desalination, 163(1-3), 253-259.
2. Tanambell, H., Danielsen, M., Freund, S., Nissen, S. H., Møller, A. H., Corredig, M., & Dalsgaard, T. K. (2024). Partitioning of proteins and small molecular weight compounds from alfalfa juice during ultrafiltration. Food Bioscience, 62, 105044.
3. Yu, Y., Kleuter, M., America, A. H., Trindade, L. M., & van der Goot, A. J. (2024). Tomato (Solanum lycopersicum) leaf juice induced whey protein gelling: Unveiling the potential of endogenous proteases in novel applications. Food Hydrocolloids, 151, 109812.