초록
<P><B>Abstract</B></P> <P>An often associated drawback with Baeyer-Villiger monooxygenases, is its poor operational stability. Furthermore, these biocatalysts frequently suffer from substrate/product inhibition.</P> <P>In this work, a thermostable cyclohexanone monooxygenase (TmCHMO) was immobilized and used in the synthesis of trimethyl-ε-caprolactone (CHL). As a cofactor regeneration enzyme, a novel and highly active glucose dehydrogenase (GDH-01) was used immobilized for the first time. MANA-agarose was the carrier chosen since it presented an immobilization yield of 76.3 ± 0.7% and a retained activity of 62.6 ± 2.3%, the highest metrics among the supports tested.</P> <P>Both immobilized enzymes were studied either separately or together in six reaction cycles (30 mL; [substrate] =132.5 mM). A biocatalyst yield of 37.3 g g<SUP>−1</SUP> of TmCHMO and 474.2 g g<SUP>−1</SUP> of GDH-01 were obtained. These values represent a 3.6-fold and 1.9-fold increase respectively, compared with a model reaction where both enzymes were used in its soluble form.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Caprolactone derivatives were synthesized using immobilized and highly active glucose dehydrogenase and thermostable BVMO as biocatalysts. </LI> <LI> When immobilized enzymes were employed, a biocatalyst yield of 37.3 g g<SUP>−1</SUP> of TmCHMO and 474.2 g g<SUP>−1</SUP> of GDH-01 were obtained. </LI> <LI> The biocatalyst yields represent a 3.6-fold and 1.9-fold increase for TmCHMO and GDH-01 respectively, compared with the use of soluble enzymes. </LI> <LI> The biocatalyst yield of GDH-01 was increased 14-fold compared with previous reported results with a GDH <I>Thermoplasma acidophilum</I>. </LI> <LI> Results represent a new input for the implementation of TmCHMO and the novel GDH-01 in the industrial production of ε-caprolactone derivatives. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>