*Article* **High Hydrostatic Pressure-Assisted Enzymatic Hydrolysis A**ff**ect Mealworm Allergenic Proteins**

**Abir Boukil <sup>1</sup> , Véronique Perreault <sup>1</sup> , Julien Chamberland <sup>1</sup> , Samir Mezdour <sup>2</sup> , Yves Pouliot <sup>1</sup> and Alain Doyen 1,\***


Academic Editors: Przemyslaw Lukasz Kowalczewski and Antonio-José Trujillo Received: 12 May 2020; Accepted: 4 June 2020; Published: 9 June 2020

**Abstract:** Edible insects have garnered increased interest as alternative protein sources due to the world's growing population. However, the allergenicity of specific insect proteins is a major concern for both industry and consumers. This preliminary study investigated the capacity of high hydrostatic pressure (HHP) coupled to enzymatic hydrolysis by Alcalase® or pepsin in order to improve the in vitro digestion of mealworm proteins, specifically allergenic proteins. Pressurization was applied as pretreatment before in vitro digestion or, simultaneously, during hydrolysis. The degree of hydrolysis was compared between the different treatments and a mass spectrometry-based proteomic method was used to determine the efficiency of allergenic protein hydrolysis. Only the Alcalase® hydrolysis under pressure improved the degree of hydrolysis of mealworm proteins. Moreover, the in vitro digestion of the main allergenic proteins was increased by pressurization conditions that were specifically coupled to pepsin hydrolysis. Consequently, HHP-assisted enzymatic hydrolysis represents an alternative strategy to conventional hydrolysis for generating a large amount of peptide originating from allergenic mealworm proteins, and for lowering their immunoreactivity, for food, nutraceutical, and pharmaceutical applications.

**Keywords:** high hydrostatic pressure; mealworm proteins; enzymatic hydrolysis; allergenic proteins; proteomics analysis
