Coding
CBDcipA

Part:BBa_K1615111

Designed by: Brooke Rothschild-Mancinelli   Group: iGEM15_Edinburgh   (2015-08-27)

CBDcipA with N and C-terminal linker in RFC25

Cellulose binding domains (CBDs) mediate the binding of enzymes to cellulose1. CBDs are divided into over a dozen families based on their sequence homology 2. Family III of CBDs is divided into a, b and c with CBDCipA belonging to family III a; the clostridial scaffoldin CBDs3. CBDCipA was identified in Clostridium thermocellum and is capable of binding to crystalline cellulose in a reversible manner4. CBDCipA includes endogenous linker sequences at both the N and C-terminals which help to prevent the CBD from interfering with the folding of any other protein it may be fused to.

Cipa graph.jpg

To characterise the binding of CBDCipA to Whatman 54 we fused it to RFP (BBa_K1615089) at both the N and C terminals and looked at dissociation of the CBD over time.


1Ferreira, L. M., Durrant, A. J., Hall, J., Hazlewood, G. P., & Gilbert, H. J. (1990). Spatial separation of protein domains is not necessary for catalytic activity or substrate binding in a xylanase. Biochem. J, 269, 261-264.
2Tomme, P., Warren, R. A. J., Miller, R. C., Kilburn, D. G. & Gilkes, N. R. (1995). Enzymatic Degradation of Insoluble Polysaccharides, edited by J. M. Saddler & M. H. Penner, pp. 142±161. Washington, DC: American Chemical Society
3Shimon, L. J., Belaich, A., Belaich, J. P., Bayer, E. A., Lamed, R., Shoham, Y., & Frolow, F. (2000). Structure of a family IIIa scaffoldin CBD from the cellulosome of Clostridium cellulolyticum at 2.2 Å resolution. Acta Crystallographica Section D: Biological Crystallography, 56(12), 1560-1568.
4Bayer, E. A., Chanzy, H., Lamed, R., & Shoham, Y. (1998). Cellulose, cellulases and cellulosomes. Current opinion in structural biology, 8(5), 548-557.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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