Coding

Part:BBa_K912000

Designed by: Grant Nicholas   Group: iGEM12_Northwestern   (2012-07-09)

Citrobacter braakii phytase

Phytase is an enzyme that breaks down phytic acid. This particular phytase was cloned from Citrobacter braakii, a gram-negative bacteria. This phytase acts to break down phytic acid into inositol, releasing phosphate groups on the phytate. If the phytate has chelated any other particle, phytase should break down the phytate to the point where the chelated particle is released.

Usage and Biology

Once expressed, the E. coli cell had to be lysed in order to release the phytase into the extra-cellular environment. We used Part:BBa_K112808 in order to lyse the cell. Once the phytase is released it is free to act upon the phytic acid.

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HokkaidoU_Japan 2017 analyzed this part. As temperature rises, this phytase's activity falls as below graph.

Protocol: 50μl 7.5 mM Phytic acid 190μl CH3COOH Buffer

Digest phytic acid in above situation with sample phytase solution 10μl, 37degrees celcius, 10 min.

After digestion, using AMM solution (2ml of 10mM NH4Mo7O2.4H2O: 5N H2SO4: Acetone= 1:1:2) analyze Optical Absorbance. The more Pi are there, the more yellow AMM becomes.

T--HokkaidoU_Japan--K912000gurafu.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1005
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 67


[edit]
Categories
//chassis/prokaryote/ecoli
//collections/probiotics/production
Parameters
None