Cofactor Regeneration in Whole-Cells

The catalytic efficiency of P450cam expressed in E. coli could be increased up to 25-fold by the coupling of its native electron transfer system (putidaredoxin reductase and putidaredoxin) to enzymatic NADH regeneration catalyzed by a recombinantly expressed glycerol dehydrogenase. This whole-cell system was applied for camphor hydroxylation in aqueous solution, as well as in a biphasic system with isooctane [181]. 

As in the case of human P450s (described in Section 12.3.2.3), a variety of artificial fusion enzymes have also been generated for bacterial P450s aiming to increase enzyme activity and coupling efficiency. 

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