title: Recombinant production and film properties of full-length hornet silk proteins.
abstract: Full-length versions of the four main components of silk cocoons of Vespa simillima hornets, Vssilk1-4, were produced as recombinant proteins in Escherichia coli. In shake flasks, the recombinant Vssilk proteins yielded 160-330mg recombinant proteinl(-1). Films generated from solutions of single Vssilk proteins had a secondary structure similar to that of films generated from native hornet silk. The films made from individual recombinant hornet silk proteins had similar or enhanced mechanical performance compared with films generated from native hornet silk, possibly reflecting the homogeneity of the recombinant proteins. The pH-dependent changes in zeta (ζ) potential of each Vssilk film were measured, and isoelectric points (pI) of Vssilk1-4 were determined as 8.9, 9.1, 5.0 and 4.2, respectively. The pI of native hornet silk, a combination of the four Vssilk proteins, was 4.7, a value similar to that of Bombyx mori silkworm silk. Films generated from Vssilk1 and 2 had net positive charge under physiological conditions and showed significantly higher cell adhesion activity. It is proposed that recombinant hornet silk is a valuable new material with potential for cell culture applications.
authors.affiliation: Silk Materials Research Unit, National Institute of Agrobiological Sciences (NIAS), 1-2 Owashi, Tsukuba, Ibaraki 305-8634, Japan.
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Kambe
authors.affiliation: Ecosystem Sciences, Commonwealth Scientific and Industrial Research Organisation (CSIRO), Clunies Ross St., Acton, ACT 2601, Australia.
authors.forename: Tara D
authors.initials: TD
authors.lastname: Sutherland
authors.affiliation: Silk Materials Research Unit, National Institute of Agrobiological Sciences (NIAS), 1-2 Owashi, Tsukuba, Ibaraki 305-8634, Japan. Electronic address: kamedat@affrc.go.jp.
authors.forename: Tsunenori
authors.initials: T
authors.lastname: Kameda
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology*; D055119:Elastic Modulus; D004926:Escherichia coli / Q000502:physiology; D008422:Materials Testing; D008567:Membranes, Artificial; D051379:Mice; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D011487:Protein Conformation; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D047011:Silk / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure*; D013314:Stress, Mechanical; D013499:Surface Properties; D013718:Tensile Strength; D014863:Wasps / Q000737:chemistry*
chemical_list: D008567:Membranes, Artificial; D011994:Recombinant Proteins; D047011:Silk
keywords: Biomimetic material; Fourier transform infrared spectroscopy; Isoelectric point; Mechanical test; Surface analysis