Highlights•Cell-free enzyme systems offer advantages over whole-cells in chemical
biomanufacturing.•Multi-enzyme systems run step-wise reactions in one-pot with unique …

Significant advances in enzyme discovery, protein and reaction engineering have
transformed biocatalysis into a viable technology for the industrial scale manufacturing of …

Single enzyme systems or engineered microbial hosts have been used for decades but the
notion of assembling multiple enzymes into cell-free synthetic pathways is a relatively new …

Industrial biocatalysis is playing a key role in the development of the global bio‐economy
that must change our current productive model to pair the socio‐economical development …

Artificial multienzyme scaffolds are being developed for in vitro cascaded biocatalytic activity
and, in particular, accessing substrate channeling. This review covers progress in this field …

Engineering biological systems for the production of biofuels and bioproducts holds great
potential to transform the bioeconomy, but often requires laborious, time-consuming design …

Although cell-free biosystems have been used as a tool for investigating fundamental
aspects of biological systems for more than 100 years, they are becoming an emerging …

Engineering microenvironments for sequential enzymatic reactions has attracted specific
interest within different fields of research as an effective strategy to improve the catalytic …

The immobilization of multi-enzyme systems on advanced materials is an emerging
technology inspired by the spatial localization found in Nature. These systems harness the …

Co-immobilization of multi-enzymes has emerged as a promising concept to design and
signify bio-catalysis engineering. Undoubtedly, the existence and importance of basic …