Biofabricating Functional Soft Matter Using Protein Engineering to Enable Enzymatic Assembly

Abstract

Biology often provides the inspiration for functional soft matter, but biology can do more: it can provide the raw materials and mechanisms for hierarchical assembly. Biology uses polymers to perform various functions, and biologically derived polymers can serve as sustainable, self-assembling, and high-performance materials platforms for life-science applications. Biology employs enzymes for site-specific reactions that are used to both disassemble and assemble biopolymers both to and from component parts. By exploiting protein engineering methodologies, proteins can be modified to make them more susceptible to biology's native enzymatic activities. They can be engineered with fusion tags that provide (short sequences of amino acids at the C- and/or N- termini) that provide the accessible residues for the assembling enzymes to recognize and react with. This "biobased" fabrication not only allows biology's nanoscale components (i.e., proteins) to be engineered, but also provides the means to organize these components into the hierarchical structures that are prevalent in life.

From stars to stripes: RNA‐directed shaping of plant viral protein templates—structural synthetic virology for smart biohybrid nanostructures

“…In combination with functional hetero‐molecules, such and similar protein species with inducible shape modulation may operate as transducer structures or contribute to morphological adaptations of smart materials. Numerous projects are addressing prospects and production routes of dynamic and adaptive nanostructured materials (Y. Liu et al, ; Luo et al, ; Rybtchinski, ; Studart, ). Hence, robust, easily manageable and well‐characterized biogenic building blocks like those derived of TMV and other plant viruses seem perfectly suited to advance this area of research further.…”

“…This is of special interest for example, for the in vivo delivery of self‐targeting diagnostic and therapeutic molecules into animals and humans, the fabrication of hand‐held devices for point‐of‐need analytics in hospitals, production facilities, supermarkets and ecosystems, or for autonomously responsive materials with sensing capacities. Numerous excellent reviews and textbooks have taken this up so that only a few ones dealing with general concepts are exemplified here (Chen, Wilmanns, & Zeng, ; Hamley, ; Hasan et al, ; Hou et al, ; Y. Liu et al, ; Luo, Hou, Bai, Wang, & Liu, ; Luo, Weiss, Liu, & Tian, ; Payne, ; Shi et al, ; L. Wang, Gong, Yuan, & Wei, ; L. Yang et al, ). Further motivations for bio‐integrative industrial process chains lie in their energetic and environmental advantages so that “bio‐based” fabrication is part of the bio‐economy strategies developed for many countries (Biber‐Freudenberger, Basukala, Bruckner, & Börner, ).…”

“…This is of special interest for example, for the in vivo delivery of self-targeting diagnostic and therapeutic molecules into animals and humans, the fabrication of hand-held devices for point-of-need analytics in hospitals, production facilities, supermarkets and ecosystems, or for autonomously responsive materials with sensing capacities. Numerous excellent reviews and textbooks have taken this up so that only a few ones dealing with general concepts are exemplified here (Chen, Wilmanns, & Zeng, 2010;Hamley, 2019;Hasan et al, 2018;Hou et al, 2017;Y. Liu et al, 2018;Luo, Hou, Bai, Wang, & Liu, 2016;Luo, Weiss, Liu, & Tian, 2018;Payne, 2007;Shi et al, 2018;L.…”

Post‐translational Assembly of Protein Parts into Complex Devices by Using SpyTag/SpyCatcher Protein Ligase

“…As this method relies on sortase catalysis, experimental conditions such as enzyme concentration can be limiting. Recent reviews describe methods to enzymatically conjugate proteins, such as by using ligases, oxidoreductases, transferases and peptidases …”

“…Recent reviewsd escribe methods to enzymatically conjugatep roteins,s uch as by using ligases, oxidoreductases, transferases and peptidases. [14][15][16][17] Split-inteins,w hichc ontain an N-terminal fragment (Int N ) and aC -terminal fragment (Int C )c an be fused to target proteins. Once reconstituted in solution, split inteins will spontaneously splice themselves out, and in the process, covalently Exploiting the innate modularity of proteins hasa llowed advancesa cross the fields of syntheticb iology and biotechnology.B yu sing standardized protein components as building blocks, complex, multiprotein assemblies with sophisticated functions can be generated;f eats previously not possible with strictly genetic-engineering approaches.…”