In
a biological system, the spatiotemporal arrangement of enzymes
in a dense cellular milieu, subcellular compartments, membrane-associated
enzyme complexes on cell surfaces, scaffold-organized proteins, protein
clusters, and modular enzymes have presented many paradigms for possible
multienzyme immobilization designs that were adapted artificially.
In metabolic channeling, the catalytic sites of participating enzymes
are close enough to channelize the transient compound, creating a
high local concentration of the metabolite and minimizing the interference
of a competing pathway for the same precursor. Over the years, these
phenomena had motivated researchers to make their immobilization approach
naturally realistic by generating multienzyme fusion, cluster formation
via affinity domain–ligand binding, cross-linking, conjugation
on/in the biomolecular scaffold of the protein and nucleic acids,
and self-assembly of amphiphilic molecules. This review begins with
the discussion of substrate channeling strategies and recent empirical
efforts to build it synthetically. After that, an elaborate discussion
covering prevalent concepts related to the enhancement of immobilized
enzymes’ catalytic performance is presented. Further, the central
part of the review summarizes the progress in nature motivated multienzyme
assembly over the past decade. In this section, special attention
has been rendered by classifying the nature-inspired strategies into
three main categories: (i) multienzyme/domain complex mimic (scaffold-free),
(ii) immobilization on the biomolecular scaffold, and (iii) compartmentalization.
In particular, a detailed overview is correlated to the natural counterpart
with advances made in the field. We have then discussed the beneficial
account of coassembly of multienzymes and provided a synopsis of the
essential parameters in the rational coimmobilization design.