Artificial Multi-Enzyme Cascades with Isolated Enzymes

1 Artificial Multi-Enzyme Cascades with Isolated Enzymes 

Artificial cascades performed with isolated enzymes in vitro (cell-free extracts or purified enzymes) provide several advantages compared to artificial cascades in recombinant cells. Multi-enzyme reactions based on isolated enzymes can easily be controlled in a desired maimer, including key factors such as enzyme combination in defined ratios and adjustment of specific reaction component ratios (i.e., cofactors). This allows a more detailed control of the biocatalytic multi-enzyme system in contrast to in vivo cascades where the above-mentioned factors are much more difficult to control. 

Two very recent examples of artificial in vitro cascade reactions combining a regio- and chemoselective P450 with an ADff operating in a simultaneous one-pot reaction mode have been reported by the groups of Groger and Schwaneberg. 

Both described biocatalytic routes employing monooxygenase-containing whole cells and isolated and purified ADHs displayed excellent regioselectivities of 99% and the intermediate alcohols were completely converted to the corresponding ketones [110], 

Multi-Enzyme Systems and Cascade Reactions Involving Cytochrome P450 Monooxygenases 

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