Difference between revisions of "Part:BBa K912000"

 
 
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<partinfo>BBa_K912000 short</partinfo>
 
<partinfo>BBa_K912000 short</partinfo>
  
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Part to produce C. braakii phytase. This phytase acts to break down phytic acid into inositol, releasing chelated iron and other micronutrients.  
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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.
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==Usage and Biology==
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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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'''Please visit our [http://2012.igem.org/Team:Northwestern wiki] for more information!'''
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HokkaidoU_Japan 2017 analyzed this part. As temperature rises, this phytase's activity falls as below graph.
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Protocol:
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50μl 7.5 mM Phytic acid
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190μl CH3COOH Buffer
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Digest phytic acid in above situation with sample phytase solution 10μl, 37degrees celcius, 10 min.
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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.
  
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<!-- Add more about the biology of this part here
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https://static.igem.org/mediawiki/2017/b/b1/T--HokkaidoU_Japan--K912000gurafu.png
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===Usage and Biology===
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Latest revision as of 03:08, 2 November 2017

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.

Please visit our [http://2012.igem.org/Team:Northwestern wiki] for more information!

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