Purification of protease enzyme from Freesia (Freesia refracta) flowers and investigation of its use in cosmetics

Abstract

The aim of this research is to identify a novel plant-based protease and explore its potential applications in the cosmetic industry. The investigation focuses on the isolation and characterisation of a protease enzyme derived from Freesia refracta flowers. The purification process involves the use of ammonium sulphate precipitation and ion exchange chromatography. In addition, the substrate specificity of the protease enzyme will be evaluated, in particular its reactions with azoalbumin and serum albumin. The enzymatic kinetics, including the determination of Vmax and Km values by Lineweaver-Burk plots, revealed values of 0.784 mg/L.min and 0.317 mM, respectively. In addition, the degree of purification and the molecular weight of the enzyme were determined by SDS-PAGE and gel filtration chromatography, with a molecular weight of 24 kDa. Based on these results, the specific type of protease enzyme obtained from Freesia flowers is identified, indicating its potential suitability for cosmetic applications. In addition, studies on the effect of various compounds such as SDS, DIPF, EDTA, phenanthraline, iodoacetamide, Ca²⁺, Zn²⁺, Hg²⁺ and Co²⁺ at a concentration of 10 mM significantly enhance the enzymatic activity of the purified protease. The research results demonstrated the successful utilization of the protease from Freesia flowers (Freesia refracta) in cosmetic formulations, demonstrating its practical applicability for cosmetic production.