Composite

Part:BBa_K118025:Experience

Designed by: Chris French   Group: iGEM08_Edinburgh   (2008-10-23)
Revision as of 00:50, 14 September 2011 by Watermelon (Talk | contribs) (→‎Characterization)

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Applications of BBa_K118025

Limonene synthase expression with pRARE2 in C41 DE3 E.coli that can be used to synthesize limonene terpenes.

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Characterization

GC-MS analysis of Limonene Synthase function

The limonene generator with lac promoter (BBa_K118025) includes the limonene generator (BBa_K118024) under the control of a lac promoter (BBa_J33207). This composite part's purpose is to express the limonene synthase (BBa_I742110), which is also available with a ribosome site (BBa_I742111).

Gas Chromatography Mass Spectrum of (+)-Limonene: We expressed the limonene generator with lac promoter (BBa_K118025) in C41 DE3 E. coli and lysed the cell culture to obtain unpurified protein extract. The crude extract was used in an in vitro enzymatic assay to produce limonene from geranyl diphosphate (GPP). The samples were prepared and analyzed by gas chromatography mass spectrometry. At retention time 8.864 minutes, the base peak at 93m/z is characteristic of limonene.