Engineering of silk proteins for materials applications

Saric, Merisa; Scheibel, Thomas

doi:10.1016/j.copbio.2019.05.005

Cited by 74 publications

(62 citation statements)

References 54 publications

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“…Consequently, the production of recombinant spidroins and their functionalisation into fibres of synthetic silk has been a research topic of significant interest. In addition, the use of synthetic biology enables the design and production of novel recombinant spidroins tailored towards specific functions in addition to those required by spiders [11]. Recombinant spidroins can also serve as model proteins in the investigation of structure-function relationships of native silk proteins [12][13][14].…”

Section: The Allure Of Spider Silkmentioning

confidence: 99%

Host Systems for the Production of Recombinant Spider Silk

Whittall

Baker

Breitling

et al. 2021

Trends in Biotechnology

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Spider silk is renowned for its impressive mechanical properties. It is one of the strongest known biomaterials, possessing mechanical properties that outmatch both steel and Kevlar. However, the farming of spiders for their silk is unfeasible. Consequently, production of recombinant spider silk proteins (spidroins) in more amenable hosts is an exciting field of research. For large-scale production to be viable, a heterologous silk production system that is both highly efficient and cost effective is essential. Genes encoding recombinant spidroin have been expressed in bacterial, yeast, insect, and mammalian cells, in addition to many other platforms. This review discusses the recent advances in exploiting an increasingly diverse range of host platforms in the heterologous production of recombinant spidroins. Highlights Recombinant spidroins of comparable size and mechanical function to native silk proteins have been successfully produced by engineered E. coli. Recombinant spidroins have been expressed in transgenic plants, rice, and alfalfa. These expression platforms can potentially offer increased industrialscale economic viability over alternative host systems.

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Section: The Allure Of Spider Silkmentioning

confidence: 99%

Host Systems for the Production of Recombinant Spider Silk

Whittall

Baker

Breitling

et al. 2021

Trends in Biotechnology

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“…Silk is a natural polymer with excellent mechanical properties, biocompatibility, and biodegradability [8]. Moreover, silk proteins have been used to form various morphological structures, such as sponges, nonwoven mats, hydrogels, films, fibers, scaffolds, capsules, and spheres [8][9][10]. Predominantly, silk biomaterials are made of silkworm silk extracted from Bombyx mori cocoons or spider silks that are mainly biotechnologically produced [9,11].…”

Section: Introductionmentioning

confidence: 99%

Drug affinity and targeted delivery: double functionalization of silk spheres for controlled doxorubicin delivery into Her2-positive cancer cells

et al. 2020

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Background: The optimal drug delivery system should be biocompatible, biodegradable, and allow the sustained release of the drug only after it reaches the target cells. Silk, as a natural polymer, is a great candidate for building drug carriers. Genetically engineered silks offer the possibility of functionalization. Previously, we characterized bioengineered silk spheres that were functionalized with H2.1 peptide that selectively delivered a drug to Her2-positive cancer cells. However, drug leakage from the silk spheres showed the need for improved control. Results: To control the drug loading and release, we designed and produced functional silk (DOXMS2) that contains a DOX peptide with an affinity for doxorubicin. The DOXMS2 spheres showed the decreased release of doxorubicin compared with MS2 particles. Next, the DOXMS2 silk was blended with the H2.1MS1 polymer to improve the control of doxorubicin binding and release into Her2-positive cancer cells. The H2.1MS1:DOXMS2 particles showed the highest doxorubicin-loading capacity and binding per cell, which resulted in the highest cytotoxic effect compared with that of other sphere variants. Since drug release at a pH of 7.4 from the blended H2.1MS1:DOXMS2 particles was significantly lower than from blended spheres without DOXMS2 silk, this indicated that such particles could control the release of the drug into the circulatory system before the carrier reached the tumor site. Conclusions: This strategy, which is based on the blending of silks, allows for the generation of particles that deliver drugs in a controlled manner.

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“…Spider silk is a kind of ideal biomaterial attributed to its extraordinary performances, such as outstanding mechanical properties and excellent biocompatibility [1][2][3]. In nature silk proteins are specifically spun into fibers, whereas after processed under various condition in vitro they can assemble into different forms with distinct morphologies, e.g.…”

Section: Introductionmentioning

confidence: 99%

One-step heating strategy for efficient solubilization of recombinant spider silk protein from inclusion bodies

Cai

Chen

et al. 2020

BMC Biotechnol

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Background: Spider silk is a proteinaceous fiber with remarkable mechanical properties spun from spider silk proteins (spidroins). Engineering spidroins have been successfully produced in a variety of heterologous hosts and the most widely used expression system is Escherichia coli (E. coli). So far, recombinantly expressed spidroins often form insoluble inclusion bodies (IBs), which will often be dissolved under extremely harsh conditions in a traditional manner, e.g. either 8 mol/L urea or 6 mol/L guanidine hydrochloride, highly risking to poor recovery of bioactive proteins as well as unexpected precipitations during dialysis process. Results: Here, we present a mild solubilization strategy-one-step heating method to solubilize spidroins from IBs, with combining spidroins' high thermal stability with low concentration of urea. A 430-aa recombinant protein (designated as NM) derived from the minor ampullate spidroin of Araneus ventricosus was expressed in E. coli, and the recombinant proteins were mainly present in insoluble fraction as IBs. The isolated IBs were solubilized parallelly by both traditional urea-denatured method and one-step heating method, respectively. The solubilization efficiency of NM IBs in Tris-HCl pH 8.0 containing 4 mol/L urea by one-step heating method was already comparable to that of 7 mol/L urea with using traditional urea-denatured method. The effects of buffer, pH and temperature conditions on NM IBs solubilization of one-step heating method were evaluated, respectively, based on which the recommended conditions are: heating temperature 70-90°C for 20 min, pH 7.0-10, urea concentration 2-4 mol/L in normal biological buffers. The recombinant NM generated via the one-step heating method held the potential functions with self-assembling into sphere nanoparticles with smooth morphology. Conclusions: The one-step heating method introduced here efficiently solubilizes IBs under relatively mild conditions compared to the traditional ones, which might be important for the downstream applications; however, this protocol should be pursued carefully in terms of urea-induced modification sensitive applications. Further, this method can be applied under broad buffer, pH and temperature conditions, conferring the potential to apply to other thermal stable proteins.

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Engineering of silk proteins for materials applications

Cited by 74 publications

References 54 publications

Host Systems for the Production of Recombinant Spider Silk

Host Systems for the Production of Recombinant Spider Silk

Drug affinity and targeted delivery: double functionalization of silk spheres for controlled doxorubicin delivery into Her2-positive cancer cells

One-step heating strategy for efficient solubilization of recombinant spider silk protein from inclusion bodies

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