Nanoformulation of cold-adapted enzymes for postharvest fungal control in export fruits

Overview

This solution offers a biological nanoformulation designed to control postharvest phytopathogenic fungi affecting export fruits. By combining cold-adapted (psychrophilic) chitinases and glucanases within a pectin polymer matrix, the formulation targets fungal cell wall components to inhibit germination of key pathogens such as Botrytis cinerea and Alternaria alternata. The approach builds on INIA's established track record in recombinant expression of antimicrobial peptides and enzymes for agricultural pest control, providing a sustainable alternative to synthetic fungicides for the fruit export industry.

Technical specifications

• Core active ingredients: Psychrophilic chitinases and glucanases, enzymes that degrade fungal cell wall polymers (chitin and glucan).
• Expression system: Recombinant production in Escherichia coli BL21 using the Gateway expression system, followed by purification from inclusion bodies and protein renaturation.
• Delivery matrix: Nanoformulation embedded in a pectin polymer matrix, enabling controlled release and compatibility with fruit surface applications.
• Target pathogens: Botrytis cinerea and Alternaria alternata, two of the most economically significant postharvest fungi in table grape, berry, and stone fruit production.
• Validation approach: In vitro inhibition assays on PDA (Potato Dextrose Agar) media followed by efficacy testing on fruit.
• Team expertise: Builds on prior work with Trichoderma-derived chitinases for botrytis tolerance and recombinant antimicrobial peptides effective against Botrytis and Alternaria spp.

Technology readiness level

The project is at an early-to-mid development stage. Heterologous expression, purification, and nanoformulation of the enzymes are planned within a 12-month project timeline. Initial proof-of-concept will be demonstrated through in vitro fungal germination inhibition assays, followed by validation on fruit. Prior team experience with related recombinant enzymes and antimicrobial peptides provides a strong foundation, though full-scale field validation and commercial formulation optimization remain to be completed.