Multienzymatic Cascades and Nanomaterial Scaffolding—A Potential Way Forward for the Efficient Biosynthesis of Novel Chemical Products

Abstract

Synthetic biology is touted as the next industrial revolution as it promises access to greener biocatalytic syntheses to replace many industrial organic chemistries. Here, it is shown to what synthetic biology can offer in the form of multienzyme cascades for the synthesis of the most basic of new materials—chemicals, including especially designer chemical products and their analogs. Since achieving this is predicated on dramatically expanding the chemical space that enzymes access, such chemistry will probably be undertaken in cell‐free or minimalist formats to overcome the inherent toxicity of non‐natural substrates to living cells. Laying out relevant aspects that need to be considered in the design of multi‐enzymatic cascades for these purposes is begun. Representative multienzymatic cascades are critically reviewed, which have been specifically developed for the synthesis of compounds that have either been made only by traditional organic synthesis along with those cascades utilized for novel compound syntheses. Lastly, an overview of strategies that look toward exploiting bio/nanomaterials for accessing channeling and other nanoscale materials phenomena in vitro to direct novel enzymatic biosynthesis and improve catalytic efficiency is provided. Finally, a perspective on what is needed for this field to develop in the short and long term is presented.

Interface Visualization of Bio-material Interaction Via Cryo-AEM Using the Biosynthesis of Iron-Based Nanoparticles as a Model

“…Nanoparticles synthesized by microorganisms have functional and economic uses and have attracted the attention of scholars in fields such as environmental remediation, functional materials, and biochemistry. − The various microscopic mechanisms of organic–inorganic interaction affect nanoparticle composition, size, morphology, surface properties, and aggregation. , Consequently, the visual analysis of the organic–inorganic interaction at the microscopic interface is of great significance for the precise engineering regulation of biosynthetic nanoparticles. The electron transport at the inorganic–organic interface is a crucial aspect of the microbial synthesis of nanomaterials. , However, due to the diversity of biosynthetic nanoparticles processes (including both intracellular and extracellular synthesis methods, incorporating complex biochemical reactions and fusion of biological organisms), subtle changes at the organic–inorganic interface and in situ characterization are challenging.…”

Enhancing enzymatic activity with nanoparticle display – an updated compendium and engineering outlook

“…1–5 There are several different approaches to achieving such enhancement, some of which have been applied independently or even jointly, and these include looking for more permissive or active enzyme homologs from other source species, mutational selection and/or evolution for optimized versions of the enzyme itself, and parametric adjustment of overall components and reaction conditions in a multienzyme cascade within a design of experimental framework. 6–16 Interestingly, attaching enzymes to macroscale scaffolds such as surfaces, beads, and resins can help increase an enzyme's structural stability and, in turn, it's viable lifetime providing for long-term application along with potential reuse by allowing for the attached enzymes to be removed with the scaffolding and added to another reaction. 17,18 However, the latter is not normally pursued to enhance enzyme activity such as the catalytic rate or k cat , for example, primarily because chemical attachment to macroscale scaffolding materials is usually achieved with a concomitant decrease in that enzyme's kinetic properties.…”

“…The potential application where we believe that NP-enzyme enhancement may play an outsized role is within designer minimalistic cell-free multienzyme cascaded reactions. 1–6,200 Rather than engineering and maintaining a cellular system to produce a desired enzymatic product, as is the current major focus in the growing field of synthetic biology, minimizing the required reactions to where all that is present are the requisite enzymes, substrates, and cofactors can allow for production of target molecules that are normally toxic to cells or otherwise not favored due to competing intracellular pathways and cross-inhibiting reactions. 201–205 Moreover, such a format can allow for potential incorporation of xenobiotic or non-natural substrates towards making new molecules enzymatically.…”

“…201–205 Moreover, such a format can allow for potential incorporation of xenobiotic or non-natural substrates towards making new molecules enzymatically. 6,200 The primary issue faced by these types of reaction formats is that of diffusion limitations and enzyme stability and it is here where NP display of enzymes can contribute. Although not focused on here, attaching enzymes to NPs can and, in many cases, does increase their long-term stability and viable lifetime along with increasing their catalytic activity.…”