Journal of the Science of Food and Agriculture · 2015 · 64 citations · 38 references
Life SciencesBiochemistryBiotechnologyEnzyme MechanismEnzymologyUltrasoundEnzyme BiophysicsEnzyme CharacterizationEnzyme KineticsEnzyme StabilityProteolytic EnzymesStructural EnzymologyCircular DichroismCircular Dichroism AnalysisCircular Dichroism Spectroscopy AnalysisMolecular Conformation AnalysisMolecular Conformation ControlProtein Conformation PreservationRandom Coil ConformationSpecific Surface Area
BACKGROUND: The disadvantages which stem from the use of traditional enzymolysis of protein has necessitated the need to employ sweeping frequency and pulsed ultrasound (SFPU) in the pretreatment of rapeseed protein prior to proteolysis in order to bring about improvement in enzymolysis efficiency. Further, in order to determine the mechanism of ultrasound-accelerated enzymolysis of RP, the effects of SFPU on the kinetics, thermodynamics, molecular conformation and microstructure of RP were investigated. RESULTS: Kinetic studies showed that SFPU pretreatment on RP improved enzymolysis by decreasing the apparent constant KM significantly (P < 0.05) by 32.8% and reducing the thermodynamic parameters Ea , ΔH and ΔS by 16.6%, 17.7% and 9.2% respectively. Fluorescence spectra revealed that SFPU pretreatment induced molecular unfolding, causing more hydrophobic groups and regions inside the molecules to be exposed to the outside. Circular dichroism analysis indicated that SFPU pretreatment decreased the α-helix content by 16.1% and increased the random coil content by 3.6%. In addition, scanning electron microscopy showed that SFPU pretreatment increased the specific surface area of RP. CONCLUSION: Ultrasound pretreatment is an efficient method in RP proteolysis to produce peptides through its impact on the molecular conformation and microstructure of proteins.
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