Purification studies of Limonene-8-hydrase from Sphingobium yanoikuyae

This thesis chapter reports an investigation into the production and purification of a key microbial enzyme involved in the biotransformation of monoterpenes into oxygenated derivatives of industrial interest. Various recovery and purification techniques were tested to isolate the enzyme from cellular material. Despite methodical efforts, the enzyme was not recovered in an active form, leading to the hypothesis that its catalytic activity depends on membrane integrity. These findings point to the likelihood that the enzyme is membrane-associated and may require detergent-assisted strategies for successful solubilization and functional characterization. The study contributes to the broader understanding of membrane-bound biocatalysts and highlights the challenges involved in their isolation for industrial applications.