Coding

Part:BBa_K118022:Experience

Designed by: Andrew Hall   Group: iGEM08_Edinburgh   (2008-10-07)
Revision as of 14:12, 13 September 2011 by Allancrossman (Talk | contribs) (→‎Characterization)

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Applications of BBa_K118022

Characterization

Edinburgh 2011 conducted two assays, comparing the activity of this part (but under the control of the lac promoter, giving BBa_K523016) to a β-glucosidase (E. coli bglX, also under the control of the lac promoter) on two different substrates:

  • 4-methylumbelliferyl β- D- glucuronide (MUG, left photo). This substrate is a cellobiose analog.
  • 4-methylumbelliferyl β- D- cellobioside (MUC, right photo). This substrate is larger and is more like a cellulose analog.

Both substrates produce a fluorescent product when cleaved. Our plates below show the results of placing cell lysate and cell debris on an MUG plate and an MUC plate. Present on both plates are:

  • Left side of plate: JM109 expressing bglX, BBa_K523014
  • Right side of plate: JM109 expressing exoglucanase cex, BBa_K523016
  • Bottom of plate: JM109 cells
BglX-MuG.jpg     Cex-MuC.jpg
MUG assay. bglX on left, cex on right.     MUC assay. bglX on left, cex on right.

As can be seen, bglX is capable of degrading MUG (the cellobiose analog) while exoglucanase displays much weaker activity. By contrast, exoglucanase is much better at degrading MUC.

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