Difference between revisions of "Part:BBa K1398003"

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https://static.igem.org/mediawiki/2014/8/86/NG_Controls_small.jpg
 
https://static.igem.org/mediawiki/2014/8/86/NG_Controls_small.jpg
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Analysis of these data allowed the kinetic parameters of Km and Vmax of NemA for nitroglycerin to be calculated as 6 M and 21 umol per mg per minute, respectively.
  
 
Full article at: http://2014.igem.org/Team:Exeter/EnzymeValidation
 
Full article at: http://2014.igem.org/Team:Exeter/EnzymeValidation

Revision as of 14:44, 17 October 2014

NemA (Inducible Construct)

A construct created by the 2014 Exeter iGEM team to degrade TNT and nitroglycerin. The construct contains the coding sequence for NemA (BBa_K1398002), an enzyme involved in the degradation of toxic compounds for their reuse in nitrogen metabolism. The construct also contains a Lactose-inducible promoter (BBa_R0010 ), a strong RBS (BBa_B0034) and a double terminator made up of BBa_B0010 and BBa_B0012. The protein has been codon-optimised for expression in E. coli.

The Exeter 2014 team used Raman spectroscopy to show the breakdown of Nitroglycerin by NemA:

NemA_degradation_of_nitroglycerin_kinetics_exp.png

The Exeter 2014 team also showed that NemA cannot perform this catalytic function without cofactors: FMN, NADH and NADPH.

NG_Controls_small.jpg Analysis of these data allowed the kinetic parameters of Km and Vmax of NemA for nitroglycerin to be calculated as 6 M and 21 umol per mg per minute, respectively.

Full article at: http://2014.igem.org/Team:Exeter/EnzymeValidation

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 NgoMIV site found at 746
    Illegal AgeI site found at 583
  • 1000
    COMPATIBLE WITH RFC[1000]