Abstract
Silks have been used widely for human beings due to their several extraordinary properties. Until now, the studies on silk proteins have mainly focused on spiders and silkworms. Because silk properties are organism-dependent, novel silk protein types can be found and developed through investigation of new silk-bearing organisms. We noticed that marine mussel has silk-like domains containing many repeats with abundance of glycine and alanine. In the present work, we redesigned mussel-derived silk-like gene sequence which contains alternating repeated and nonrepeated regions with optimized codons for Escherichia coli. For successful expression of recombinant mussel silk-like protein in E. coli cells, we employed several experimental strategies, including use of strong promoter, cold shock expression, and genetic fusions. We observed significant repression on cell growths by even low expression levels of soluble mussel silk-like proteins in cold shock- and glutathione s-transferase (GST) fusion-based systems. Thus, we finally used baculoviral polyhedrin protein as a fusion partner and successfully expressed insoluble mussel silk-like protein with relatively high expression level and without cell growth repression in E. coli.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Y. Cao and B. Wang, Int. J. Mol. Sci., 10, 1514 (2009).
J. M. Gosline, M. E. Demont and M.W. Denny, Endeavour, 10, 37 (1986).
R. V. Lewis, Chem. Rev., 106, 3762 (2006).
J. Huang, C. Wong Po Foo and D. L. Kaplan, Polym. Rev., 47, 29 (2007).
T. D. Sutherland, J. S. Church, X. Hu, M.G. Huson, D. L. Kaplan and S. Weisman, PLoS ONE, 6, e16489 (2011).
S. Weisman, V. S. Haritos, J. S. Church, M.G. Huson, S. T. Mudie, A. J.W. Rodgers, G. J. Dumsday and T. D. Sutherland, Biomaterials, 31, 2695 (2010).
J. Gosline, M. Lillie, E. Carrington, P. Guerette, C. Ortlepp and K. Savage, Phil. Trans. R. Soc. Lond., B 357, 121 (2002).
C. Vendrely and T. Scheibel, Macromol. Biosci., 7, 401 (2007).
M. J. Harrington and J. H. Waite, J. Exp. Biol., 210, 4307 (2007).
X.-X. Qin, K. J. Coyne and J. H. Waite, J. Biol. Chem., 272, 32623 (1997).
X.-X. Qin and J. H. Waite, Proc. Natl. Acad. Sci., 95, 10517 (1998).
K. J. Coyne, X.-X. Qin and J. H. Waite, Science, 277, 1830 (1997).
S. J. Lombardi and D. L. Kaplan, J. Arachnol., 18, 297 (1990).
A. Rising, M. Widhe, J. Johansson and M. Hedhammar, Cell. Mol. Life Sci., 68, 169 (2011).
C. L. Craig and C. Riekel, Comp. Biochem. Phys., B., 133, 493 (2002).
P. Guerette, D. Ginzinger, B. Weber and J. Gosline, Science, 272, 112 (1996).
H. G. Menzella, Microb. Cell Fact., 10, 15 (2011).
S. Winkler and D. L. Kaplan, Rev. Mol. Biotechnol., 74, 85 (2000).
S. R. Fahnestock and L.A. Bedzyk, Appl. Microbiol. Biotechnol., 47, 33 (1997).
P. E. Vaillancourt, E. coli Gene Expression Protocols, 205, 1 (2003).
J. Goldstein, N. S. Pollitt and M. Inouye, Proc. Natl. Acad. Sci., 87, 283 (1990).
H. Tanabe, J. Goldstein, M. Yang and M. Inouye, J. Bacteriol., 174, 3867 (1992).
J. Steczko, G.A. Donoho, J. E. Dixon, T. Sugimoto and B. Axelrod, Protein Expr. Purif., 2, 221 (1991).
M. Tamura, K. Ito, S. Kunihiro, C. Yamasaki and M. Haragauchi, Protein Expr. Purif., 78, 1 (2011).
S. C. Makrides, Microbiol. Rev., 60, 512 (1996).
E. R. LaVallie and J. M. McCoy, Curr. Opin. Biotechnol., 6, 501 (1995).
W. Guo, L. Cao, Z. Jia, G. Wu, T. Li, F. Lu and Z. Lu, Protein Expr. Purif., 77, 185 (2011).
K. A. Harrap, Virology, 50, 124 (1972).
G. F. Rohrmann, J. Gen. Virol., 67, 1499 (1986).
J. H. Seo, L. Li, J. S. Yeo and H. J. Cha, Biotechnol. Bioeng., 84, 467 (2003).
R. Rudolph and H. Lilie, FASEB J., 10, 49 (1996).
R. L. Quant, M. N. Pearson, G. F. Rohrmann and G. S. Beaudreau, Appl. Environ. Microbiol., 48, 732 (1984).
Q. Wei, Y. S. Kim, J.H. Seo, W. S. Jang, I. H. Lee and H. J. Cha, Appl. Environ. Microbiol., 71, 5038 (2005).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Yang, Y.J., Choi, Y.S., Jung, D. et al. Expression of redesigned mussel silk-like protein in Escherichia coli . Korean J. Chem. Eng. 28, 1744–1748 (2011). https://doi.org/10.1007/s11814-011-0140-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11814-011-0140-3