Sterol side-chain cleavage with immobilized Mycobacterium cells in water-immiscible organic solvents

The possibility of using whole Mycobacterium sp. cells for the selective degradation of the side-chain of sitosterol in an organic bioconversion medium was investigated. Sterol solubility limits were estimated and free-cell biocompatibility tests were carried out, using a range of water-immiscible solvents. Among these, phthalates exhibited good biocompatibility and sterol-solubilizing capacities. Several organic and inorganic matrices were tested for the immobilization of Mycobacterium sp. cells by surface adhesion. Celite led to the best results, being thus selected for beta-sitosterol side-chain degradation tests in phthalates and in aqueous medium. Cells entrapped in kappa-carrageenan and in polyurethane foams were also used in these tests. The highest degradation activities were obtained with cells immobilized in Celite, with bis(2-ethylhexyl)phthalate as the conversion medium, resulting in molar conversion yields up to 70%, with respect to beta-sitosterol (5 g l-1). Further activity and stability tests revealed that this bioconversion system is markedly temperature dependent.