Coding
Part:BBa_K4580002:Design
Designed by: Michael Constant Group: iGEM23_Cornell (2023-10-09)
Enzyme Isolation
To allow for easy enzyme isolation, a 6x-histidine tag was added to the NdmD. This design choice allowed Team Cornell to use Ni-NTA Affinity Chromatography to isolate our enzymes of interest for further encapsulation into our reactor.
Source
NdmA, NdmB, and, NdmD were amplified from the bacteria Pseudomonas putida CBB5 genomic DNA.
References
Summers, R. M., Louie, T. M., Yu, C. L., Gakhar, L., Louie, K. C., & Subramanian, M. (2012). Novel, highly specific N-demethylases enable bacteria to live on caffeine and related purine alkaloids. Journal of bacteriology, 194(8), 2041–2049. https://doi.org/10.1128/JB.06637-11
UT Austin. (2012). BBA_K734000. NdmABCD operon. https://parts.igem.org/Part:BBa_K734000