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IMPROVEMENT OF FUNCTIONAL AND BIOACTIVE PROPERTIES OF CHIA SEED (SALVIA HISPANICA) PROTEIN HYDROLYSATES AND DEVELOPMENT OF BIODEGRADABLE FILMS USING CHIA SEED MUCILAGE
Chia seed (Salvia hispanica) has shown potential as an alternative source of nutrients with a high content of fiber (36 %), protein (25%), and fat (25%). Unfortunately, the presence of a viscous biopolymer (mucilage), surrounding the chia seed (CS), limits the accessibility of the protein and other nutrients. Nevertheless, this biopolymer’s chemical composition makes it suitable for the development of biodegradable films. Regarding CS protein, disulfide bonding, and nonprotein-protein interactions often frequent in plant protein, have limited its technological application in food matrices. Therefore, scientists have pointed at processing methods involving enzymatic proteolysis to improve the functionality of plant protein ingredients. The objective of this study was to establish processing techniques to exploit the functionality, extraction, and health benefits of chia seed components. First, ultrasonication followed by vacuum-filtration was used to separate mucilage from CS prior to fat extraction by oil press. Mucilage-free and defatted CS were treated using conventional (enzymatic hydrolysis with alcalase) or sequential (enzymatic hydrolysis with alcalase+flavourzyme), and under water bath or microwave-assisted hydrolysis. Chia seed protein hydrolysates (CSPH) derived from the sequential hydrolysis with microwave treatment showed superior (p<0.05) in vitro antioxidant activity. The highest (p<0.05) cellular antioxidant activity was achieved by the sequential (94.76%) and conventional (93.13%) hydrolysis with microwave. Dipeptidyl peptidase-V inhibition was higher (p<0.05) for sequential hydrolysis with water bath, while Angiotensin-Converting Enzyme (ACE) inhibition activity increased (p<0.05) with hydrolysis for all treatments compared to the control. Regarding functionality, sequential hydrolysis with microwave showed higher (p<0.05) solubility at lower pH (3 and 5), while conventional hydrolysis with microwave was better at pH 7 and 9. Emulsification properties and foaming capacity were also higher in conventional hydrolysis with microwave, but conventional hydrolysis with water bath was more stable for foaming properties only. In terms of mucilage applicability, biodegradable films were developed by casting technique where CS mucilage was plasticized with different polyol mixtures (sorbitol and glycerol). CS mucilage films with higher sorbitol content showed superior tensile strength (3.23 N/mm2), and lower water vapor permeability (1.3*109 g/ m*s*Pa) but had poor flexibility compared to other treatments. Conversely, films with high glycerol content showed high elongation at break (67.55%) and solubility (22.75%), but reduced water vapor permeability and tensile strength. The hydrophobicity, measured as water contact angle, was higher (p<0.05) for mixtures containing equal amounts of polyols. Lastly, Raman Spectroscopy analysis showed shifts from 854 to 872 cm-1 and 1061 to 1076 cm-1, which corresponded to β(CCO) modes. These shifts represent an increase in hydrogen bonding, responsible for the high tensile strength and decreased water vapor permeability. This study demonstrated that ultrasonication followed by vacuum filtration can successfully separate mucilage from chia seeds; microwave-assisted and enzymatic hydrolysis generated protein hydrolysates with improved bioactivity and functionality. Finally, chia seed mucilage was able to form films with potential to be used in drug delivery and edible food coating applications.