Emerging Solutions for in Vivo Biocatalyst Immobilization: Tailor-Made Catalysts for Industrial Biocatalysis

Abstract

In industry, enzymes are often immobilized to generate more stable enzyme preparations that are easier to store, handle, and recycle for repetitive use. Traditionally, enzymes are bound to inorganic carrier materials, which requires caseto-case optimization and incurs additional labor and costs. Therefore, with the advent of rational protein design strategies as part of bottom-up synthetic biology approaches, numerous immobilization methods have been developed that enable the one-step production and immobilization of enzymes onto biogenic carrier materials often directly within the production host, which we here refer to as in vivo immobilization. As a result, nano-to micro-meter-sized functionalized biomaterials, or biologically produced enzyme immobilizates, are obtained that can directly be used for synthetic purposes. In this Perspective, we provide an overview over established and recently emerging in vivo enzyme immobilization methods, with special emphasis on their applicability for (industrial) biocatalysis. For each approach, we present fundamental working principles as well as advantages and limitations guiding future research avenues toward sustainable applications in the bioindustry.

Combining a Genetically Engineered Oxidase with Hydrogen‐Bonded Organic Frameworks (HOFs) for Highly Efficient Biocomposites

“…[ 6 , 7 , 8 , 9 , 10 ] Like most non‐fibrous proteins, enzymes are inherently fragile ex situ, hindering their use, for example, in commercial catalysis where recyclability is desired. [ 11 , 12 , 13 ] Thus, immobilization on, or within, solids is employed as a strategy to enhance enzyme stability in catalysis, biomedical science and biosensing applications. [ 6 , 8 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 ] For example, infiltration into preformed porous materials (e. g., silica, organic polymers) is well known for enzyme immobilization.…”

Methodological advances and strategies for high resolution structure determination of cellular protein aggregates

“…Consequently, IBs are being explored for applications as drug delivery systems and self-immobilized catalysts. IBs formed by enzymes, such as reductases, kinases, lipases, and synthases, have the advantage of catalyzing reactions while being reusable and recyclable, resistant to proteases, and relatively easy to purify ( 8 , 141 , 142 , 143 ). In all these occurrences of aggregation, structural and mechanistic knowledge would be an asset, contributing to the development of disease treatment, biotherapeutics, and catalysts in industrial processes, for example ( 144 , 145 , 146 ).…”

Putting precision and elegance in enzyme immobilisation with bio-orthogonal chemistry

“…Primarily aimed at further streamlining of biocatalytic processes, a multitude of methods for the cost-effective in vivo immobilisation of enzymes have been developed in recent years 20 by combining enzyme production with in vivo immobilisation to afford a one-step, cost-effective production of an immobilised enzyme. This is achieved by genetic fusion of a self-assembly promoting protein to the target enzyme.…”