Enzyme immobilization has been extensively employed in research and industry to improve enzyme stability and allow enzyme recycling. Broad ranges of chemicals and support materials have been utilized for enzyme immobilization. Recent breakthroughs in nanotechnology and materials science have influenced enzyme immobilization technology. Novel approaches for enzyme immobilization have enabled us to access more benefits, for example, excellent activity and stability, cost effectiveness, and the establishment of continuous flow processes. Herewith, this review provides an update on the enzyme immobilization strategies that have been attracting expanding interest in biocatalysis. Organic–inorganic nanocrystal formation, metal–organic frameworks, enzyme immobilization on graphene-based nanomaterials, and enzyme immobilization on functionalized solid surfaces are discussed in detail.

Geotrichum candidum acetophenone reductase (GcAPRD) nanocrystal reduces broad kinds of ketones to their corresponding (S)-alcohols with excellent enantioselectivity.

Enantiopure alcohol production by a flow system of 3D-printed bioreactor with immobilized Geotrichum candidum acetophenone reductase (GcAPRD).

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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 (FBVMO), was isolated from a soil sample. FBVMO was overexpressed in E. coli and successfully immobilized by the organic-inorganic nanocrystal formation method. The immobilization improved the thermostability of FBVMO. Substrate specificity investigation revealed that both free and immobilized FBVMO 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 FBVMO 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.