Enzyme evolution for industrial biocatalytic cascades
Multi-step, biocatalytic cascades are poised to lead to further adoption of enzymes by the chemical industry. Over the past twenty years, the promise of in vitro enzyme evolution for the sustainable biocatalytic synthesis of complex chemicals at large scale has materialized. Recently, the ﬁeld of biocatalysis is seeing further expansion, with biocatalytic processes becoming more complex and involving multiple consecutive enzymatic conversions. These biocatalytic cascades are assembled in single reaction vessels to accomplish difﬁcult chemistry under mild reaction conditions, with minimal waste generation and attractive economics. Advances in enzyme engineering have enabled the increasingly efﬁcient optimization of enzymes in the context of such cascades, where each enzyme operates in the presence of others, under continuously changing conditions as substrate, reaction intermediates, and product concentrations ﬂuctuate over the course of the reaction. Enzyme evolution has provided biocatalysts with greatly improved traits, including activity, selectivity, and stability. This review focuses on recently developed, industrially relevant enzyme cascades.