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Microalgae are a valuable source of enzymes and active ingredients due to their biochemical variability, health-beneficial properties, lower production cost and viability on an industrial scale. In addition, the growing demand for sustainable products constitutes microalgae as a viable resource for the development of a range of innovative products. In the present work, the proteolytic activity of <i>Arthrospira platensis</i> lysate was assessed and characterized using kinetics analysis and zymography employing different substrates, pH values and divalent ions. The results suggest the presence of two main proteolytic enzymes in the lysate. In addition, a rapid and easy purification protocol of the proteolytic activity, under mild conditions, was developed using a polyethyleneglycol (PEG)/phosphate aqueous two-phase system (ATPS). The proteolytic activity of <i>A. platensis</i> lysate was used to develop a hydrogel formulation as an enzyme-based cosmeceutical, with potential application as a topical exfoliating agent. The incorporation of the <i>A. platensis</i> extract in the developed hydrogel formulation significantly improved its operational stability over time, which is a significant advantage in enzyme-based product development.