Immobilization of Baeyer-Villiger monooxygenase from acetone grown Fusarium sp.

Abstract

ObjectiveA novel biocatalyst for Baeyer–Villiger oxidations is necessary for pharmaceutical and chemical industries, so this study aims to find a Baeyer–Villiger monooxygenase (BVMO) and to improve its stability by immobilization. ResultsAcetone, the simplest ketone, was selected as the only carbon source for the screening of microorganisms with a BVMO. A eukaryote, Fusarium sp. NBRC 109816, with a BVMO ( F BVMO), was isolated from a soil sample. F BVMO was overexpressed in E. coli and successfully immobilized by the organic-inorganic nanocrystal formation method. The immobilization improved the thermostability of F BVMO. Substrate specificity investigation revealed that both free and immobilized F BVMO were found to show catalytic activities not only for Baeyer–Villiger oxidation of ketones to esters but also for oxidation of sulfides to sulfoxides. Furthermore, a preparative scale reaction using immobilized F BVMO was successfully conducted. ConclusionsFBVMO was discovered from an environmental sample, overexpressed in E. coli , and immobilized by the organic-inorganic nanocrystal formation method. The immobilization successfully improved its thermostability.

Recent Advances in Enzyme Immobilization Utilizing Nanotechnology for Biocatalysis

“…This approach is cost-effective and straightforward, so numerous functional proteins have been immobilized by nanocrystal formation and extensively applied in broad study areas 15,27,29 such as biosensors, 30 bioremediation, 31 biomedicine, 32 bioassays, 33 wastewater treatment, 34 and industrial biocatalysis. 27,35 Various enzymes have been immobilized by nanocrystal formation, including lipase, 27,36−40 peroxidase, 41−43 laccase, 27,41 glucoamylase, 44 protease, 45 papain, 46 alcohol dehydrogenase, 47,48 aldehyde dehydrogenase, 29 Baeyer−Villiger monooxygenase, 49 urease, 50 and esterase. 51 This section presents the current outstanding features of nanocrystal formation: the mechanism of the nanocrystal formation, variability in the nanocrystal formation process, enzyme activity enhancement, multipurpose nanocrystal formation that simultaneously achieves both immobilization and purification of enzymes, and potential applications.…”