@inproceedings{01JK5FGSXYC1MG5PECHQPVEGJ9,
  abstract     = {{The Farm to Fork strategy outlined in the EU Green Deal aims to slash chemical pesticide usage by 50% by 2030, a pivotal step in promoting sustainable crop protection. Despite this ambitious initiative, nearly half of total pesticide usage consists of fungicides, highlighting the urgent necessity for bio-based alternatives. In response to this challenge, ProTECTzymes investigates the potential of engineered antifungal enzymes as bio-based solutions. This approach seamlessly aligns with the objectives of the EU Green Deal and holds promise for ensuring high-quality agriculture for future generations.

We selected a chitinase and a beta-1,3-glucanase that target the structural polymers of the fungal cell wall—chitin and beta-1,3-glucan, respectively. To assess their direct lytic effect, a microscopy study was conducted with non-engineered enzyme mixtures. This in vitro study revealed a direct lytic effect on germinating spores of Botrytis cinerea, termed the first mode of action. Subsequently, tomato leaf disks were infected with B. cinerea, and after treatment, infection progression was monitored using multispectral plant imaging. The results demonstrated a significant reduction in infection symptoms and a deceleration of infection progression. Additionally, the activation of the plant's innate immune system was visualized, indicating a second mode of action by the generated chitin-oligomers and glucan-oligomers, acting as PAMPs.

Following this initial proof-of-concept, the antifungal properties of the enzyme mixtures were enhanced by converting them into engineered fusion proteins, coined ProTECTzymes. The introduced colocalization effect potentially enhances their cooperative activity by placing the catalytic domains at the same place, at the same time. After selecting for expression yields, ProTECTzymes were tested using leaf disk assays, revealing heightened antifungal activity compared to the free enzyme mixtures.

The implications of our results are profound. This proof-of-concept study underscores the tremendous potential of enzyme-based fungicides. Beyond their immediate antifungal properties, the integration of synthetic biology allows for optimization, enabling efficacy at low concentrations. This optimization process further opens avenues for potentially tailoring proTECTzymes to be pathogen-specific, addressing a critical need in precision agriculture. Moreover, the biodegradability of these enzyme-based fungicides positions them as environmentally friendly alternatives, paving the way for greener production in agrobiotechnology.}},
  author       = {{Meert, Kenan and Drijvers, Fien and Perneel, Maaike and Briers, Yves and Audenaert, Kris}},
  booktitle    = {{Plant BioProTech & IOBC-IR 2024, Abstracts}},
  keywords     = {{protein,enzyme,biofungicide,synthetic biology}},
  language     = {{eng}},
  location     = {{Castellon de la Plana, Spain}},
  title        = {{ProTECTzymes : synthetic fungal cell wall-degrading enzymes as biofungicides}},
  year         = {{2024}},
}