An official website of the United States government
Renewable and degradable materials, formed using biopolymers as material precursors, are sought after in pharmaceutical, biomedical, and industrial fields. Silk-based biomaterials, primarily derived from the silk fibroin protein of the Bombyx mori (B. mori) silkworm, have advantageous mechanical properties, biocompatibility, and commercial availability. Recent efforts aim to expand the range of achievable silk-based biomaterial properties via alternative sources of silk proteins with different sequences and structures. These structural distinctions drive differences in physical and chemical properties of silk fibers, primarily due to the varying degree of crystallinity in the polymers. For the development of alternative silk-based materials, silk from Plodia interpunctella (P. interpunctella), a small agricultural pest that infests and damages food products via silk production, is evaluated. Early investigations have highlighted differences between P. interpunctella and B. mori silk fibroin proteins, however P. interpunctella silk still largely lacks characterization and optimization on both the silk fiber and bulk material level. This work evaluates the structural, thermal, mechanical, and cell-material properties of non-degummed and degummed P. interpunctella silk as a raw material for biomaterial fabrication and discusses the benefits and limitations of these proteins as new biopolymers. Observed properties are used to identify links between silk fibroin protein sequence and fiber function in addition to forming hypotheses in how P. interpunctella silk-based biomaterials will perform in comparison to other natural biopolymers. Future work aims to develop methods to process P. interpunctella silk into material formats, utilizing the material characteristics determined here as a baseline for shifts in material performance.
more »
« less
Secretion‐Catalyzed Assembly of Protein Biomaterials on a Bacterial Membrane Surface
Abstract Protein‐based biomaterials have played a key role in tissue engineering, and additional exciting applications as self‐healing materials and sustainable polymers are emerging. Over the past few decades, recombinant expression and production of various fibrous proteins from microbes have been demonstrated; however, the resulting proteins typically must then be purified and processed by humans to form usable fibers and materials. Here, we show that the Gram‐positive bacteriumBacillus subtiliscan be programmed to secrete silk through its translocon via an orthogonal signal peptide/peptidase pair. Surprisingly, we discover that this translocation mechanism drives the silk proteins to assemble into fibers spontaneously on the cell surface, in a process we call secretion‐catalyzed assembly (SCA). Secreted silk fibers form self‐healing hydrogels with minimal processing. Alternatively, the fibers retained on the membrane provide a facile route to create engineered living materials fromBacilluscells. This work provides a blueprint to achieve autonomous assembly of protein biomaterials in useful morphologies directly from microbial factories.
more »
« less
- Award ID(s):
- 2045844
- PAR ID:
- 10482659
- Publisher / Repository:
- Wiley
- Date Published:
- Journal Name:
- Angewandte Chemie International Edition
- Volume:
- 62
- Issue:
- 37
- ISSN:
- 1433-7851
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
More Like this
-
-
Silk fibers are produced by a wide variety of insects. The silkworm Bombyx mori (Bombyx) was domesticated because the physical properties of its silk fibers were amenable to the production of fine textiles. Subsequently, engineers have regenerated silk fibroin to form biomaterials. The monocular focus on Bombyx silk has underutilized the expanse of diverse silk proteins produced by more than 100,000 other arthropods. This vast array of silk fibers could be utilized for biomedical engineering challenges if sufficient rearing and purification processes are developed. Herein, we show that the moth, Plodia interpunctella (Plodia), represents an alternative silk source that is easily reared in highly regulated culture environments allowing for greater consistency in the silk produced. We controlled the temperature, resource availability (larvae/gram diet), and population density (larvae/mL) with the goal of increasing silk fiber production and improving homogeneity in Plodia silk proteins. We determined that higher temperatures accelerated insect growth and reduced life cycle length. Furthermore, we established initial protocols for the production of Plodia silk with optimal silk production occurring at 24 °C, with a resource availability of 10 larvae/gram and a population density of 0.72 larvae/mL. Population density was shown to be the most prominent driving force of Plodia silk mat formation among the three parameters assessed. Future work will need to link gene expression, protein production and purification, and resulting mechanical properties as a function of environmental cues to further transition Plodia silk into regenerated silk fibroin biomaterials.more » « less
-
Abstract BackgroundThe increasing prevalence of plastic waste combined with the inefficiencies of mechanical recycling has inspired interest in processes that can convert these waste streams into value-added biomaterials. To date, the microbial conversion of plastic substrates into biomaterials has been predominantly limited to polyhydroxyalkanoates production. Expanding the capabilities of these microbial conversion platforms to include a greater diversity of products generated from plastic waste streams can serve to promote the adoption of these technologies at a larger scale and encourage a more sustainable materials economy. ResultsHerein, we report the development of a new strain ofPseudomonasbacteria capable of converting depolymerized polyethylene into high value bespoke recombinant protein products. Using hexadecane, a proxy for depolymerized polyethylene, as a sole carbon nutrient source, we optimized media compositions that facilitate robust biomass growth above 1 × 109 cfu/ml, with results suggesting the benefits of lower hydrocarbon concentrations and the use of NH4Cl as a nitrogen source. We genomically integrated recombinant genes for green fluorescent protein and spider dragline-inspired silk protein, and we showed their expression inPseudomonas aeruginosa, reaching titers of approximately 10 mg/L when hexadecane was used as the sole carbon source. Lastly, we demonstrated that chemically depolymerized polyethylene, comprised of a mixture of branched and unbranched alkanes, could be converted into silk protein byPseudomonas aeruginosaat titers of 11.3 ± 1.1 mg/L. ConclusionThis work demonstrates a microbial platform for the conversion of a both alkanes and plastic-derived substrates to recombinant, protein-based materials. The findings in this work can serve as a basis for future endeavors seeking to upcycle recalcitrant plastic wastes into value-added recombinant proteins.more » « less
-
Spider silk is biocompatible, biodegradable, and rivals some of the best synthetic materials in terms of strength and toughness. Despite extensive research, comprehensive experimental evidence of the formation and morphology of its internal structure is still limited and controversially discussed. Here, we report the complete mechanical decomposition of natural silk fibers from the golden silk orb-weaver Trichonephila clavipes into ≈10 nm-diameter nanofibrils, the material's apparent fundamental building blocks. Furthermore, we produced nanofibrils of virtually identical morphology by triggering an intrinsic self-assembly mechanism of the silk proteins. Independent physico-chemical fibrillation triggers were revealed, enabling fiber assembly from stored precursors “at-will”. This knowledge furthers the understanding of this exceptional material's fundamentals, and ultimately, leads toward the realization of silk-based high-performance materials.more » « less
-
Ma, Li-Jun (Ed.)Abstract Spiders amplify their physical capabilities by synthesizing multiple high performing silks. Renowned for its toughness, major ampullate (MA) silk composes the spiderweb frame, providing support and absorbing high-energy impacts. In ecribellate orb-weavers, proline-rich motifs in MaSp2 proteins of MA silk are linked to a range of mechanical properties, including extensibility, elasticity, stiffness, and supercontraction. We show a modification of these motifs outside of this clade in a spider that constructs a spring-loaded web. The triangle weaver spider Hyptiotes cavatus (family Uloboridae) stores energy in the support lines of its triangular web, then rapidly releases the tension to catapult forward, collapsing the web around prey. Hyptiotes has an expanded set of MaSp2 genes which encode proteins with far higher proline contents than typical MaSp2. The predominant GPGPQ motifs present in Hyptiotes spidroins also occur abundantly in MaSp sequences of distantly related spiders that produce the most extensible dragline, implying silk protein convergence. Proline-rich MaSp2 proteins constitute half of all MA gland expression in Hyptiotes, and we show that the resulting fibers are the most proline-rich spider silk measured to date. This unique silk composition suggests a functional importance that may facilitate the spring-loaded prey capture mechanism of this species' web and may inspire the design of novel biomaterials using protein engineering.more » « less
- Free Publicly Accessible Full Text
-
Accepted Manuscript
- Journal Article:
- https://doi.org/10.1002/anie.202305178
