The employment of alcohol dehydrogenases in cascade reactions is often limited by the different cofactor specificity of the enzymes involved: the employment of additional cofactor regeneration systems and the excess amount of sacrificial substrates frequently increase the environmental impact and the costs of biocatalytic processes. Additionally, a NADP+-dependent process is generally less desirable, inasmuch this cofactor is more expensive, unstable and less naturally available than NAD+, leading to an increase of the process costs.
Nowadays, protein engineering offers the possibility to switch the cofactor dependency of enzymes introducing few targeted mutation1.
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