Cascades of Evolved Enzymes for the Synthesis of Complex Molecules

Abstract

Thanks to advances in enzyme discovery and protein engineering combined with the development of enzymatic multistep reaction cascades, new efficient routes for drug synthesis have been created that are superior to chemical syntheses. This supports the goal of the chemical and pharmaceutical industries to move to more sustainable and environmentally friendly processes. Recently described outstanding examples include the biocatalytic cascade syntheses of the cyclic dinucleotide MK-1454, molnupiravir, and islatravir, as well as the efficient fixation of CO 2 to make starch using an artificial enzyme cascade.

Macrocyclization and Backbone Rearrangement During RiPP Biosynthesis by a SAM-Dependent Domain-of-Unknown-Function 692

“…Many of the post-translational modification reactions are unique chemical transformations resulting in macrocycles and heterocycles, structures that are privileged in bioactive scaffolds. Furthermore, exciting recent developments in biocatalysis have increasingly shown the value of enzymatic transformations in preparation of compounds that are useful to human society, with many of the enzymes coming from biosynthetic pathways to natural products. − Therefore, discovery of new chemical reactions catalyzed by enzymes and assignment of function to poorly characterized enzyme families is an important goal.…”

The E factor at 30: a passion for pollution prevention

“…186 Most enzymatic reactions are conducted under roughly the same conditions of temperature and pressure and, hence, it is relatively easy to integrate multiple steps into eco-efficient catalytic cascade processes. 187–189 thus avoiding the separation, and purification of the products from individual steps characteristic of conventional processing. This also improves conversions by limiting product inhibition.…”

Enzyme Cascade with Horseradish Peroxidase Readout for High-Throughput Screening and Engineering of Human Arginase-1

“…This one-pot single-step assay continuously generates a colorimetric or fluorescent signal, is compatible with a broad range of HRP substrates, and allows straightforward fitting of enzyme parameters K m and k cat without the need for performing endpoint measurements at different time points . An efficient enzymatic reaction cascade design prevents rate-limiting reactions and intermediate accumulation while providing alternative chemical routes and signal amplification. − Additionally, bacterial cell-surface display of active hArg1 was achieved and used to validate the assay. We used this assay to screen a small curated library of 13 hArg1 variants and detected small differences in K M and k cat .…”