Abstract
The demand for sustainable and natural alternatives in the flavor and fragrance industries has driven interest toward enzymatic synthetic routes. This study reports the enzymatic production of geranyl benzoate, a monoterpenoid ester with potential bioactivity and commercial applications, catalyzed by immobilized lipases using a solvent-free system. A central composite design was employed to optimize the key process variables: molar ratio, temperature, and enzyme loading using Lipozyme 435 as the biocatalyst. Under optimal conditions (1:7 molar ratio, 80°C, 30% enzyme), a high geraniol conversion (72%) was achieved within 6 hr, as validated by a predictive model. The study also investigated the inhibitory effects of coproduced methanol, demonstrating that open-reactor systems (enabling methanol evaporation) enhanced conversion (>99% in 48 h) compared to closed systems. Additionally, substrate-exposure experiments highlighted methyl benzoate's inhibitory effect on Lipozyme 435, reducing its activity by 54%, whereas Lipura Flex showed greater resilience. Reusability tests indicated a decline in enzyme performance, with Lipozyme 435 retaining 22% activity after five cycles. Cell tests demonstrated no cytotoxicity and proliferative effect at high concentrations. These findings highlight the potential of solvent-free enzymatic processes for sustainable production of geranyl benzoate and provide insights into operational challenges, including methanol management and biocatalyst stability.