Engineered a novel pH-sensitive short major ampullate spidroin

Zhang, Chen; Mi, Jiali; Qi, Haishan; Huang, Jie; Liu, Simin; Zhang, Lei; Fan, Daidi

doi:10.1016/j.ijbiomac.2020.03.153

Cited by 12 publications

(13 citation statements)

References 36 publications

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“…Concentrated small complete mini-spidroins similar to N-R 7 -C have been shown to form protein micelles in solution 9,30 . Recombinant spidroins featuring a spider eggcase silk repetitive region and both terminal domains Figure 3.…”

Section: Resultsmentioning

confidence: 99%

“…We selected a pH drop to 5.0 for comparison with this work, and have more thoroughly investigated the effects of ionic strength on this spinning process, demonstrating this to be an important factor in this approach. The development of a spinning process utilising multiple smaller pH drops, or a continuous gradient could offer further improvements 3,30,36,41,42,67 . In the development of such an approach it would likely be insightful to further consider the isoelectric point (pI) of the mini-spidroins and Supplementary Fig.…”

Section: Biomimetic Wet Spinning Of a Complete Mini-spidroin Results mentioning

confidence: 99%

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The effect of terminal globular domains on the response of recombinant mini-spidroins to fiber spinning triggers

Finnigan

Roberts

Ligorio

et al. 2020

Sci Rep

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Spider silk spidroins consist of long repetitive protein strands, flanked by globular terminal domains. The globular domains are often omitted in recombinant spidroins, but are thought to be essential for the spiders' natural spinning process. Mimicking this spinning process could be an essential step towards producing strong synthetic spider silk. Here we describe the production of a range of minispidroins with both terminal domains, and characterize their response to a number of biomimetic spinning triggers. Our results suggest that mini-spidroins which are able to form protein micelles due to the addition of both terminal domains exhibit shear-thinning, a property which native spidroins also show. Furthermore, our data also suggest that a pH drop alone is insufficient to trigger assembly in a wet-spinning process, and must be combined with salting-out for effective fiber formation. With these insights, we applied these assembly triggers for relatively biomimetic wet spinning. This work adds to the foundation of literature for developing improved biomimetic spinning techniques, which ought to result in synthetic silk that more closely approximates the unique properties of native spider silk. Spider dragline silk has impressive mechanical properties, with high strength and good extensibility resulting in a level of toughness which exceeds all other natural and synthetic fibers 1. However, unlike silkworms, spiders cannot be efficiently farmed for their silk 2. For this reason, the production of recombinant spider silk proteins (spidroins), and their subsequent spinning into synthetic spider silk fibers, has been an active topic of research for a number of decades 3-7. Major ampullate spider silk proteins (spidroins) are typically 200-350 kDa in size and constitute the dragline silk of spiders. Generally, spidroins have three distinct regions (Fig. 1) 3. The vast majority of the protein is repetitive, consisting of alternating polyalanine regions and glycine-rich regions 8. At the terminals of the spidroin exist non-repetitive domains, referred to as the N-and C-terminal domains (NTD and CTD). These globular terminal domains are crucial in facilitating the soluble storage of the spidroins at high concentrations (30-50% w/v) in the silk gland, and in initiating fiber assembly 9. Much of the research into recombinant spider silk has focused on spidroins consisting of only the repetitive region. Whilst some of the largest recombinant spidroins have resulted in fibers with good mechanical properties, larger repetitive regions typically result in poor spidroin yields 10,11. Commonly, denaturing conditions have been employed in either the purification or spinning processes. Silk proteins purified under such denaturing conditions have been shown to lack the response to shear exhibited by native spinning dopes-an essential feature to provide alignment of the fiber microstructure, as well as an assembly trigger itself 12. In contrast, biomimetic spinning utilising correctly folded terminal domains may offer the production...

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Section: Resultsmentioning

confidence: 99%

Section: Biomimetic Wet Spinning Of a Complete Mini-spidroin Results mentioning

confidence: 99%

The effect of terminal globular domains on the response of recombinant mini-spidroins to fiber spinning triggers

Finnigan

Roberts

Ligorio

et al. 2020

Sci Rep

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show abstract

“…Recombinant fibers from chimeric spidroins NT MaSp -Rep MaSp -CT MiSp (44 kDa) showed a tensile strength of about 50 MPa but a high extensibility of ∼90%. 50 The tensile strength of synthetic fibers from chimeric spidroins NT MaSp -2Rep MaSp -CT MiSp (33 kDa) reached 162 MPa and these silk fibers had an extensibility of 37%. 29 The customized FlSp silk-like fibers in the current study revealed an outstanding tensile strength comparable to the wet-spinning silk-like fibers, but a relatively lower extensibility, which was not expected.…”

Section: Discussionmentioning

confidence: 99%

“… 30 Chimeric spidroins NT MaSp -Rep-CT MiSp at 100–300 mg/mL were spun into silk-like fibers from pH 2.0 to 11. 50 Chimeric spidroins W AcSp 2C MaSp 4CT MaSp and W AcSp 2C MaSp 8CT MaSp formed silk-like fibers by hand-pulling at pH 7.5 at rather low concentration (0.4 mg/mL). Recently, silk-like fibers were pulled manually from NT MaSp R MaSp/FlSp nCT MaSp (n indicates the number of R in combination of MaSp and FlSp repeats) 11 and from NT MaSp W AcSp nCT MiSp at concentrations of (0.8–1.0 mg/mL) at pH 8.0.…”

Section: Discussionmentioning

confidence: 99%

Customized Flagelliform Spidroins Form Spider Silk-like Fibers at pH 8.0 with Outstanding Tensile Strength

Cai

et al. 2021

ACS Biomater. Sci. Eng.

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Spider flagelliform silk shows the best extensibility among various types of silk, but its biomimetic preparation has not been much studied. Herein, five customized flagelliform spidroins (FlSps: S and NTD Fl -Sn-CTD Fl , n = 1–4), in which the repetitive region (S) and N-/C- terminal domains (NTD Fl and CTD Fl ) are from the same spidroin and spider species, were produced recombinantly. The recombinant spidroins with terminal domains were able to form silk-like fibers with diameters of ∼5 μm by manual pulling at pH 8.0, where the secondary structure transformation occurred. The silk-like fibers from NTD Fl -S4-CTD Fl showed the highest tensile strength (∼250 MPa), while those ones with 1–3 S broke at a similar stress (∼180 MPa), suggesting that increasing the amounts of the repetitive region can improve the tensile strength, but a certain threshold might need to be reached. This study shows successful preparation of flagelliform silk-like fibers with good mechanical properties, providing general insights into efficient biomimetic preparations of spider silks.

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“…Further, the b-turn is partly involved in the formation of b-fold and is parallel to the fiber axis, b-turn, and a-helix. 3,4,14 According to research, the ion, 15 pH, 16,17 and temperature 18 are the factors that influence the conformation. As we know, the structure has a decisive influence on performance.…”

Section: Introductionmentioning

confidence: 99%

Progress in the application of spider silk protein in medicine

et al. 2021

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Spider silk protein has attracted much attention on account of its excellent mechanical properties, biodegradability, and biocompatibility. As the main protein component of spider silk, spidroin plays important role in spider spinning under natural circumstances and biomaterial application in medicine as well. Compare to the native spidroin which has a large molecular weight (>300 kDa) with highly repeat glycine and polyalanine regions, the recombinant spidroin was maintained the core amino motifs and much easier to collect. Here, we reviewed the application of recombinant spider silk protein eADF4(C16), major ampullate spidroin (MaSp), minor ampullate spidroin (MiSp), and the derivatives of recombinant spider silk protein in drug delivery system. Moreover, we also reviewed the application of spider silk protein in the field of alternative materials, repairing materials, wound dressing, surgical sutures along with advances in recombinant spider silk protein.

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Engineered a novel pH-sensitive short major ampullate spidroin

Cited by 12 publications

References 36 publications

The effect of terminal globular domains on the response of recombinant mini-spidroins to fiber spinning triggers

The effect of terminal globular domains on the response of recombinant mini-spidroins to fiber spinning triggers

Customized Flagelliform Spidroins Form Spider Silk-like Fibers at pH 8.0 with Outstanding Tensile Strength

Progress in the application of spider silk protein in medicine

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