Difference between revisions of "Part:BBa K801093:Experience"

Latest revision as of 02:56, 29 October 2013

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UNIQ64665e703586db0b-partinfo-00000000-QINU UNIQ64665e703586db0b-partinfo-00000001-QINU

Team UBC 2013

Team UBC 2013 used the 4- coumarate Ligase characterized by TU_Munich 2012 in a biosynthetic pathway that generated Cinnamaldehyde from the aromatic amino acid Phenylalanine. The data Below outlines the success of this enzyme in our biosynthetic pathway.

The genes encoding PAL (BBa_K1129003), 4-Coumarate CoA ligase(BBa_K1129042) and a Cinnamoyl Co-A reductase(BBa_1129005) were assembled into one PSB1C3 plasmid under constitutive promoters and transformed into E. coli. E.coli 10G cells were grown over night before being incubated with L-Phenylalanine for 7 hours, the sample was extracted before it was run on GC-MS.

Const._EncP_%2B_AtCCR1.jpg

Figure 1. Compound generation identification by GC-MS. Chromatograms (left) and mass spectra (right) for select peaks are shown. Structures represent predictions based on library matching or comparison to standards. Controls represent plasmids missing the gene of interest. Top) GC-MS of a Cinnamic acid (10.580) control. Bottom) Conversion of Phenylalanine to Cinnamaldehyde (11.819) through the cinnamic acid intermediate(10.580) via the EncP, 4CL, and ATCCR1 gene construct under a constitutive promoter. The mass spec data shows a highly oxidized cinnamaldehyde that may have resulted from poor sample preparation

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 Team UBC 2013 used the 4- coumarate Ligase characterized by TU_Munich 2012 in a biosynthetic pathway that generated Cinnamaldehyde from the aromatic amino acid Phenylalanine. The data Below outlines the success of this enzyme in our biosynthetic pathway.
-The genes encoding PAL (BBa_K1129003), 4-Courmarate CoA ligase(BBa_K1129042) and a Cinnamoyl Co-A reductase(BBa_1129005) were assembled into one PSB1C3 plasmid under constitutive promoters and transformed into E. coli. E.coli 10G cells were grown over night before being incubated with L-Phenylalanine for 7 hours, the sample was extracted before it was run on GC-MS.
+The genes encoding PAL (BBa_K1129003), 4-Coumarate CoA ligase(BBa_K1129042) and a Cinnamoyl Co-A reductase(BBa_1129005) were assembled into one PSB1C3 plasmid under constitutive promoters and transformed into E. coli. E.coli 10G cells were grown over night before being incubated with L-Phenylalanine for 7 hours, the sample was extracted before it was run on GC-MS.
 <p align=center>https://static.igem.org/mediawiki/2013/2/2a/Const._EncP_%2B_AtCCR1.jpg</p>
 '''Figure 1'''. Compound generation identification by GC-MS. Chromatograms (left) and mass spectra (right) for select peaks are shown. Structures represent predictions based on library matching or comparison to standards. Controls represent plasmids missing the gene of interest. Top) GC-MS of a Cinnamic acid (10.580) control. Bottom) Conversion of Phenylalanine to Cinnamaldehyde (11.819) through the cinnamic acid intermediate(10.580) via the EncP, 4CL, and ATCCR1 gene construct under a constitutive promoter.  The mass spec data shows a highly oxidized cinnamaldehyde that may have resulted from poor sample preparation