Difference between revisions of "Part:BBa K517002:Experience"
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==Characterization by British Columbia iGEM 2011== | ==Characterization by British Columbia iGEM 2011== | ||
| − | ===GC-MS analysis of | + | ===GC-MS analysis of Alpha-Pinene Synthase function by British Columbia iGEM 2011=== |
| − | [[Image: | + | [[Image:Ubcigem2011alphapingcms.jpg | frame | center | '''Gas Chromatography Mass Spectrum of Alpha-pinene Synthase Products:''' We expressed the alpha-pinene synthase (<partinfo>BBa_K517002</partinfo>) 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.]] |
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Revision as of 21:04, 23 September 2011
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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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UNIQc776923415dad175-partinfo-00000000-QINU UNIQc776923415dad175-partinfo-00000001-QINU
Characterization by British Columbia iGEM 2011
GC-MS analysis of Alpha-Pinene 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.