Part:BBa_K558007
Bay Laurel Thioesterse Generator for Fatty Acid Production
Bay Laurel Thioesterase is controlled by the constitutive promoter sigma 70 (J23101)with a RBS, and the double terminator (BBa_B0014) Bay Laurel Thioesterase (BTE) will intercept acyl-ACP from the fatty acid synthesis cycle and turn it into the fatty acid derivatives C12 and C14.
L Yuan, T A Voelker, and D J Hawkins, (1995), Modification of the substrate specificity of an acyl-acyl carrier protein thioesterase by protein engineering.
Proc Natl Acad Sci U S A. November 7; 92(23): 10639–10643.
Welch et. al. “Design Parameters to Control Synthetic Gene Expression in Escherichia coli”
Results:
Bay Laurel Thioesterase was tested in NEB 10 Beta cells for initial quantification. NEB 10 Beta cells showed a positive increase of 115uM of fatty acids compared to negative controls of 40uM and 70uM. NEB Iq cells transformed with Bay Laurel Thioesterase were then tested because the NEB Iq cells are a higher protein producing strain of E. coli. NEB Iq cells transformed with Bay Laurel Thioesterase had positive increase of 125uM compared to the negative controls of 50uM and 40uM
EnzyChrom Free Fatty Acid Assay Kit (BioAssay Systems)was used to determine concentration of BTE samples and negative controls.
Concentration(uM)of BTE samples vs. negative controls
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 503
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 360
Illegal BsaI.rc site found at 770
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