Microbial biofabrication for nanomedicine: biomaterials, nanoparticles and beyond

Abstract

NanomedicineVol. 8, No. 12 EditorialFree AccessMicrobial biofabrication for nanomedicine: biomaterials, nanoparticles and beyondEsther Vázquez & Antonio VillaverdeEsther VázquezInstitut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193 Barcelona, Spain and Department de Genètica i de Microbiologia, Universitat Autònoma de Barcelona, Bellaterra, 08193 Barcelona, Spain and CIBER en Bioingeniería, Biomateriales y Nanomedicina, Bellaterra, 08193 Barcelona, SpainSearch for more papers by this author & Antonio Villaverde* Author for correspondenceInstitut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Bellaterra, 08193 Barcelona, Spain and Department de Genètica i de Microbiologia, Universitat Autònoma de Barcelona, Bellaterra, 08193 Barcelona, Spain and CIBER en Bioingeniería, Biomateriales y Nanomedicina, Bellaterra, 08193 Barcelona, Spain. Search for more papers by this authorEmail the corresponding author at antoni.villaverde@uab.catPublished Online:26 Nov 2013https://doi.org/10.2217/nnm.13.164AboutSectionsPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareShare onFacebookTwitterLinkedInRedditEmail Keywords: biomaterialgenetic designmetabolic circuitmicrobial factorynanoparticleupstream engineeringUnder the generic umbrella of biotechnology and being one of its emerging facets, the concept of biofabrication comprises a set of methodologies addressed to the production of rather complex constructs with predefined biological properties. Biofabrication uses biological materials, ranging from molecules to cells, as building blocks for organized products, by using sophisticated manufacturing instruments [1]. The paradigm of biofabrication is organ printing, in which 3D organ constructs are generated by robotic technologies from preformed cell clusters [2].Although in a broader sense, controlled biological synthesis could be equated to biofabrication, the particular driving force underlying biofabrication is distinct from the plain production of macromolecules or materials (e.g., secondary metabolites, polymers, nucleic acids or proteins), as it involves a level of complexity of the final product that requires an accurate design at the upstream level. Biological production of molecules and macromolecules in cell factories (mainly microbes) supported the birth and further growth of food, industrial and pharmaceutical biotechnology. In fact, the history of modern medicine has been intimately linked to the biological synthesis of antibiotics by fungi and soil bacteria, which developed as a solid industry during the 1940s and 1950s [3]. Many other high-value chemicals, peptides and amino acids are obtained as natural products by exploiting the metabolic diversity of microorganisms and by finding optimal conditions to specifically enhance the biosynthesis of these molecules, by either process or metabolic engineering. Furthermore, in the last 30 years, and strongly supported by the development of recombinant DNA technologies, a wide spectrum of proteins with biomedical interest (including enzymes, hormones, antibodies or antigens/immunogens) have been produced at a laboratory scale for biomedical research. This has been critical for the determination of protein structure–function relationships, the biological dissection of the immune system, the progressive understanding of cell biology and signaling events, as well as for the development of interactomics and related fields. In addition, the enrichment of the recombinant DNA toolkit and the implementation of protein production as a robust generic platform in biosciences have also endorsed the integrated comprehension of regulatory circuits in life and led to questions regarding how biological components should be applied or manipulated in molecular therapies. In this regard, recombinant proteins are pivotal analytical tools. Under a different perspective, approximately 200 recombinant protein species have been approved by medicament agencies and are commercialized in the form of drugs or vaccine components for the treatment or prophylaxis of an increasing number of human diseases [4].When exploring the literature in the field, it is evident how the structural and biological complexity of different products resulting from microbial production is steadily increasing. Primarily encouraged by the need for novel functional biomaterials in emerging medicines (nanostructured or not), large catalogs of natural or engineered substances are being incorporated to the biomedical landscape, with particular value in nanomedical applications. These include polymers, metal nanoparticles, magnetic nanoparticles, virus-like particles, virions or virion components and a growing diversity of de novo designed self-organizing protein materials, some of them with regulatable biomechanical properties such as stiffness, elasticity, adhesiveness or controllable disintegration or release of functional building blocks or embedded drugs [5]. The production of most of these materials is the result of a refined designing scheme and a precise adjustment of the cell factory machinery to reach the desired functional and/or structural intricacy of the products. Importantly, this is also progressively turning biological production into a more competitive alternative of the less versatile chemical synthesis [6].The evolution from plain biotechnological production towards microbial biofabrication can be exemplified by recent progresses in the generation of smart nanoparticles addressed to medical applications. In this context, different natural protein cages, including virus-like particles, bacterial microcompartments and eukaryotic vaults, are being produced in recombinant cell factories upon refined protein engineering to grant them with specific cellular tropism or the ability to load therapeutic cargoes, such as nucleic acids or conventional chemical drugs [7]. In addition, protein domains from natural oligomers can be incorporated into heterologous protein stretches to control their self-assembling as protein nanoparticles [7]. In a further step, multifunctional monomeric proteins resulting from a fully de novo genetic design recruit a spectrum of biological activities that empower them for specific cell surface receptor binding, internalization and intracellular trafficking to reach a target compartment [8,9]. The incorporation of architectonic peptide tags to these constructs (namely arginine- and histidine-rich cationic peptides) confers an important extent of control over the size and geometry of the resulting nanoparticles and allows the manipulation of their abilities to bind cargo DNA [9]. Bacterial inclusion bodies formed by therapeutic proteins have recently been proposed as biomimetics of functional amyloids acting in the endocrine secretory system, and adapted for the controlled intracellular release of protein drugs in the form of spherical, submicron protein particles [10,11]. By genetic manipulation of the quality control system of the producing bacteria and by adjusting conditions of the biofabrication process, critical properties of these protein particles can be defined in advance [12]. In a pioneering example, the downstream conjugation of functional peptides to the material’s surface has also recently opened a route for targeted administration of nanopills to the pathogenic bacteria Helicobacter pylori[13]. In a different field, nonprotein bacterial nanoparticles can be functionalized during biofabrication by upstream genetic design. Magnetic nanoparticles produced by magnetotactic bacteria display-specific surface ligands upon generating fusion proteins with the natural transmembrane proteins (Mms13 and others) that are embedded into the lipidic membrane surrounding the iron oxide deposit itself. A similar strategy has been adopted to functionalize the surface of polyhydroxyalkanoate granules for decoration with metals in imaging or for cell targeting [5].Biofabrication, cell factories & nanomedicineTwo main strategies allow us to consider an increasing number of microbial nanoparticles and biomaterials for biomedical uses. First, the continuous screening of unconventional microbial species with valuable metabolic properties [14] leads to the production of novel or improved materials of a microbial cell size scale, that is, at a submicron level. On the other hand, the expansion of metabolic engineering to the systems level, that is, systems biotechnology, allows the fine reprograming of cells’ metabolic circuits to favor the production and accumulation of desired products, through cost-effective processes applicable at the industrial scale [15]. This results in much more functionally robust and genetically stable cell factories, in which the whole complexity and potency of the biosynthetic machineries are recruited in a systems-regulated way.Both approaches are exemplified by the recent success in the microbial biofabrication of metal (and magnetic) particles with biomedical interest, which are to be used in nanoelectronics, drug delivery, imaging, hyperthermia, as antimicrobial agents and in regenerative medicine, among other applications [16,17]. The increasing use of microbial metal nanoparticles in nanotechnology and nanomedicine results from the identification of new microbial species and consortia from natural environments [18,19]. The inclusion of novel species as cell factories expands the catalog of metal-based materials that can be obtained by biofabrication in nanostructured forms. However, this might pose potential challenges in the adaptation of these microbial species to large-scale cultivation. The use of novel biofactories may also not permit the straightforward application of genetic engineering techniques because of the unavailability of appropriate tools. Subsequently, the quality and properties of the products could be only adjusted by process engineering. By contrast, the fine metabolic manipulation of the well-known organisms, such as the enterobacterium Escherichia coli, through conventional recombinant DNA technology has recently resulted into the production of a spectrum of metallic nanoparticles with regulatable size and tailored optical, electronic, chemical and magnetic properties [20]. Again, in this regard, the systems biotechnology and reprogramming of existing microbial capabilities offers an unexpected approach to convert plain biotechnological production into biofabrication of complex nanostructured entities, by the use of well-known biological systems.ConclusionThe combination of systems biotechnology and industrial microbiology has allowed the fast adaptation of microbial cell factories from the plain bioproduction of molecules of biotechnological and biomedical interest to the biofabrication of smart biomaterials, nanoparticles and sophisticated self-organizing constructs useful in nanomedicine, whose structural and functional complexity demand a precise upstream design. In addition, the screening of unconventional microbes in natural environments has expanded the catalog of nanostructured products that can be obtained by microbial synthesis. Both classical genetic engineering and more complex systems-level reprograming of microbial metabolic routes open an avenue for exploitation of microbial diversity and biosynthetic potential in biofabrication for nanomedicine, which could, in turn, result in cost-effective, fully scalable and versatile alternatives to chemical synthesis.Financial & competing interests disclosureThe authors are indebted to MINECO (grant BFU2010-17450), FISS (PI12-00327), Agència de Gestió d’Ajuts Universitaris i de Recerca (grant 2009SGR-108), Institució Catalana de Recerca i Estudis Avançats (through and Institució Catalana de Recerca i Estudis Avançats Academia award to A Villaverde) and to The Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine for supporting their research in biomaterials and nanomedicine. 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The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.No writing assistance was utilized in the production of this manuscript.PDF download