Antifreeze protein solution for ice crystal control in ice cream

Overview

This solution offers a newly designed antifreeze protein (AFP) engineered for higher ice-binding activity than wild-type proteins. When incorporated into ice cream formulations, the AFP prevents large ice crystal formation and ice recrystallization that typically cause gritty texture during freezing, storage, and temperature fluctuations. The result is a smoother mouthfeel, reduced iciness perception, and extended shelf life, even under challenging supply chain conditions.

A key differentiator is the use of a safe microbial yeast expression system for production, which lowers manufacturing costs and supports a clean-label ingredient profile by eliminating the need for artificial chemical preservatives. This makes the technology attractive to ice cream manufacturers seeking natural, consumer-friendly ingredients that do not compromise on product quality or stability.

Technical specifications

Production and purification:

• The AFP gene is synthesized and cloned into a yeast expression vector, then transformed into a selected yeast host capable of intracellular and extracellular expression
• Standard fermentation protocols are followed, with expression confirmed by ELISA and Western blot
• Column chromatography purification ensures the AFP is contaminant-free and suitable for food applications
• Scalable production is supported by an in-house bioreactor with capacity up to 30 liters

Characterization methods:

• Ice recrystallization inhibition (IRI) assay and thermal hysteresis (TH) assay using cryomicroscopy to quantify ice-binding performance
• Differential scanning calorimetry (DSC) to assess thermal properties and freeze-thaw resilience
• Cytotoxicity evaluation through zebrafish embryo toxicity testing and MTT assays on cell lines to confirm consumer safety

Design approach:

The AFP variant (AFPIV) was engineered through chimerism with another AFP. Molecular dynamics simulations identified a mutant with the highest interaction energy with ice, longest hydrogen bond lifetime, and lowest crystal growth rate, providing a strong rationale for its superior ice-binding capability.

Technology readiness level

The AFP has been designed and characterized at the laboratory scale, with ice-binding activity validated through IRI, TH, and DSC analyses. Cytotoxicity testing has been performed to support food-grade safety. The research team (EMTech) at Universiti Putra Malaysia has demonstrated expertise across expression, purification, and formulation characterization, though access to dedicated AFP characterization facilities is currently limited and represents a potential collaboration need. Production is positioned for scale-up to a 30-liter bioreactor, indicating readiness for pilot-scale trials and formulation optimization with industry partners.