Development And Optimization Of Automated Fibre Production For Recombinant Spider Wrapping Silk
Date
2018-05-23T17:24:24Z
Authors
Weatherbee-Martin, Nathan
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Abstract
Spiders produce up to seven different types of silk, each serving different biological
functions. Wrapping silk, used for wrapping of prey, is the toughest of the seven types.
Limited information is available on wrapping silk fibre formation and structure. Native
wrapping silk protein (AcSp1) in Argiope trifasciata consists of a core domain with at least 14
consecutive 200 amino acid repeat (“W”) units. A recombinant 3 W-unit protein (W3) was
produced in Escherichia coli. Automated fibre production (“spinning”) was achieved by
solubilization of W3 in two different solvent-based spinning “dopes”, followed by extrusion,
dehydration, and post-spin stretching. Post-spin stretching improved fibre strength, enriched
β-sheet conformation, and increased birefringence. Additionally, fibre extensibility differed
with spinning dope composition, corresponding to differences in anisotropy of secondary
structuring, and in birefringence. This high-throughput synthetic recombinant spider silk
fibre production methodology will make possible studies of its atomic-level structuring using
nuclear magnetic resonance spectroscopy.
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Keywords
wrapping silk, AcSp1, bacteria, recombinant protein, spinning dope, wet-spinning, mechanical testing, Raman spectroscopy, birefringence