Cascades in Compartments: En Route to Machine‐Assisted Biotechnology

Abstract

Biological compartmentalization is a fundamental principle of life that allows cells to metabolize, propagate, or communicate with their environment. Much research is devoted to understanding this basic principle and to harness biomimetic compartments and catalytic cascades as tools for technological processes. This Review summarizes the current state-of-the-art of these developments, with a special emphasis on length scales, mass transport phenomena, and molecular scaffolding approaches, ranging from small cross-linkers over proteins and nucleic acids to colloids and patterned surfaces. We conclude that the future exploration and exploitation of these complex systems will largely benefit from technical solutions for the integrated, machine-assisted development and maintenance of a next generation of biotechnological processes. These goals should be achievable by implementing microfluidics, robotics, and added manufacturing techniques supplemented by theoretical simulations as well as computer-aided process modeling based on big data obtained from multiscale experimental analyses.

Self-assembling all-enzyme hydrogels for biocatalytic flow processes

“…It is evident from natural examples that compartmentalized reaction systems offer advantages when intermediate species are prone to escape into competing reaction channels [3][4][5][6][7][8] . The GDH-ST/SC-LbADH hydrogels can be used to investigate this phenomenon because the NADPH consuming LbADH reduction is spatially and kinetically coupled to the NADPH regeneration by GDH.…”

“…Towards the goal of the efficient use of renewable biomass as an alternative to petrochemical synthesis for sustainable production processes and energy supply in the future, multistep enzymatic cascade reactions are currently attracting much attention [3][4][5][6][7][8] . Such cascades are abundant in living systems, for instance, to maintain the regulation of metabolic activity or signal transduction [9][10][11] .…”

“…The development of biocatalytic flow chemistry-like systems is underway with a special focus on microfluidic bioreactors that offer the generic advantage of a high degree of control over temperature profiles and diffusion-based mixing 8,[16][17][18] . However, biocatalytic processes are difficult to realize in flow systems because the heterogeneous catalysis regime calls for effective surface immobilization techniques that are more demanding for enzymes than for conventional organo(metallic) catalysts 19 .…”

DNA Surface Technology: From Gene Sensors to Integrated Systems for Life and Materials Sciences

“…Them ethod is very mild because protein hybridization is achieved under physiological conditions that do not harm sensitive tertiary and quaternary structures of even complex proteins.O wing to the high surface-coating density,multiplexing capabilities,and various options to regenerate the underlying DNAsurface by simple denaturation protocols,n umerous research papers have meanwhile demonstrated the utility and scope of the DDI method. [32] Also important is the fact that non-natural DNAanalogues,such as peptide nucleic acids (PNA), can be effectively used for DDI approaches,a s demonstrated in proteomics applications. [30] Conductive silicon substrates have also been used for protein and proteome analyses.…”

Efficient synthesis of the key chiral alcohol intermediate of Crizotinib using dual‐enzyme@CaHPO4 hybrid nanoflowers assembled by mimetic biomineralization

“…In the present work, the mimetic biomineralization for preparing artificial bone–mimetic composite was used to assemble the enzymes with calcium phosphate to form the immobilized dual enzymes biocatalyst. Aldehyde‐ketone reductase and alcohol dehydrogenase were co‐crystalized with CaHPO 4 at 4 °C to form dual‐enzyme‐inorganic hybrid nanoflowers, which also takes advantage of CaHPO 4 biocompatibility and substrate channeling effects from the dual‐enzyme catalytic cascade reaction . Subsequently, the resulting assembled hybrid nanoflowers immobilized enzymes were applied in biocatalytic synthesis of the chiral alcohol intermediate of Crizotinib using ketone as substrate.…”