Coding

Part:BBa_K2160000

Designed by: Emma Finlayson-Trick   Group: iGEM16_Dalhousie_Halifax_NS   (2016-10-14)


Beta-1,4-D-glycosidic bonds in cellulose -> cellulobiose

The endoglucanase enzyme was taken from the thermophillic bacteria Ruminiclostridium thermocellum. The endoglucanase enzyme cleaves internal Beta-1,4-D-glycosidic bonds in the cellulose crystal to release the disaccharide cellulobiose.


===Usage and Biology===

Contribution

Group: Dalhousie iGEM 2017

Author: Jacob Sicheri

Summary: Our team added a C-terminal HIS-tag and a N-terminal PelB sequence to this part. The HIS-tag allows for comformation of expression, and the PelB sequence traffics the protein to the periplasm. Included below is a western blot probing for HIS-tag showing the confirmed expression of a 46 kDa species, the predicted migration of the optimized endoglucanase enzyme. Endoglucoptimized.png

Find our optimized biobrick here: https://parts.igem.org/Part:BBa_K2331004

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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 231
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None