Part:BBa_K517002:Experience

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Applications of BBa_K517002

Alpha-pinene synthase expression with pRARE2 in C41 DE3 E.coli that can be used to synthesize alpha-pinene.

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Characterization by British Columbia iGEM 2011

GC-MS analysis of Limonene Synthase function by British Columbia iGEM 2011

Gas Chromatography Mass Spectrum of Alpha-pinene Synthase Products: We expressed the alpha-pinene synthase (BBa_K517002) 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 alpha-pinene from geranyl diphosphate (GPP) substrate. The samples were prepared and analyzed by gas chromatography mass spectrometry. (A) We analysed the products of the alpha-pinene synthase and found alpha-pinene, beta-pinene and small quantities of other monoterpenes. (B) Cross referencing with standards and a compound library revealed that each monoterpene product had the characteristic base peaks and retention time. The left panel shows the identification of alpha-pinene and the right panel shows the identification of beta-pinene as products of our alpha-pinene synthase.