Coding

Part:BBa_K5281011

Designed by: Yiming Dong   Group: iGEM24_HS-Alpha   (2024-10-01)


CPR


Cytochrome P450 Reductase (CPR) is a coding gene in the S. cerevisiae,which is essential for the activity of cytochrome P450 enzymes. CPR provides the necessary electrons for P450 enzymes to carry out the oxidation reactions. This electron transfer is crucial for enabling the P450-catalyzed hydroxylation of sesquiterpenes, facilitating the conversion of santalene and bergamotene into santalols and bergamotol. More experimental results can be found on the webpage BBa_K5281017.

Verifying the expression of CYP/CPR in S. Cerevisiae

Figure 1. CYP/CPR homologous recombination verification with Colony PCR and Gel Electrophoresis

We also verified the expression of CYP736A167 and SaCPR2 with SDS-PAGE. The kDa of the two enzymes are listed below:

CYP- 55.91 kDa

CPR- 77.65 kDa

Figure 2. CYP/CPR Enzyme Expression verification with SDS PAGE

 

 

References

Zhang, J., Wang, X., Zhang, X., Zhang, Y., Wang, F., & Li, X. (2022).Sesquiterpene Synthase Engineering and Targeted Engineering of α - Santalene Overproduction in Escherichia coli.Journal of Agricultural and Food Chemistry, 70(17), 5377–5385.

Xinrui, D., Bo, L., Yihong, B., Weifeng, L., & Yong, T. (2023).Metabolic engineering of Escherichia coli for high - level production of violaxanthin.Microbial Cell Factories, 22(1).

Diaz - Chavez, M.L., Moniodis, J., Madilao, L.L., Jancsik, S., Keeling, C.I., Barbour, E.L., Ghisalberti, E.L., Plummer, J.A., Jones, C.G., & Bohlmann, J. (2013).Biosynthesis of Sandalwood Oil: Santalum album CYP76F Cytochromes P450 Produce Santalols and Bergamotol.PLoS ONE, 8(9), e75053.

Etchegaray, J., & Inouye, M. (1999).CspA, CspB, and CspG, Major Cold Shock Proteins of Escherichia coli, Are Induced at Low Temperature under Conditions That Completely Block Protein Synthesis.Journal of Bacteriology, 181(6), 1827–1830.

Kim, S.H. (2018).TIRF - Based Single - Molecule detection of the RECA presynaptic filament dynamics.Methods in Enzymology on CD - ROM / Methods in Enzymology, 233–253.

Ahn, J.M., Mitchell, R.J., & Gu, M.B. (2004).Detection and classification of oxidative damaging stresses using recombinant bioluminescent bacteria harboring sodA∷, pqi∷, and katG∷luxCDABE fusions.Enzyme and Microbial Technology, 35(6–7), 540–544.

Wang, Y., Ling, N., Jiao, R., Zhang, X., Ren, Y., Zhao, W., Chen, H., & Ye, Y. (2024).A universal mechanism on desiccation tolerance of Cronobacter based on intracellular trehalose accumulation regulated by EnvZ / OmpR.Food Microbiology, 119, 104455.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 804
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 562
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 106


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