Coding

Part:BBa_K4033004

Designed by: Chenxi Wang   Group: iGEM21_BHSF   (2021-08-24)
Revision as of 04:12, 25 August 2021 by XiXiThea (Talk | contribs)

DnrF for coding

The DnrF gene, responsible for conversion of aklavinone to ɛ-rhodomycinone via C-11 hydroxylation, was mapped in the daunorubicin (Dnr) gene cluster of Streptomyces peucetius ATCC 29050. The DnrF gene was sequenced and should encode a protein of 489 amino acids with a molecular mass of 52 kDa.[1] It is used as monooxygenase in our design which can finally let dcll become carminic acid.

Biology and Usage

DnrF involved in the biosynthesis of the anthracyclines carminomycin, rhodomycin, daunorubicin (daunomycin) and doxorubicin (adriamycin) which are aromatic polyketide antibiotics that exhibit high cytotoxicity and are widely applied in the chemotherapy of a variety of cancers. Catalyzes the incorporation of a hydroxyl group at position C-11 of aklavinone, resulting in epsilon-rhodomycinone.[2]

DnrF can convert aklabinone to ɛ-rhodomycinone via C-11 hydroxylation. The heterologous expression of DnrF in Escherichia coli demonstrated that the gene encodes a NADPH-dependent hydroxylase catalysing the hydroxylation of aklavinone to yield ɛ-rhodomycinone. It is used as monooxygenase in our design which can finally let dcll become carminic acid.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 508
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 508
    Illegal NotI site found at 1219
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1074
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 508
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 508
    Illegal AgeI site found at 722
  • 1000
    COMPATIBLE WITH RFC[1000]


Experimental approach



References

[1] Filippini, S & Solinas, M & Breme, U & Schlüter, M & Gabellini, D & Biamonti, Giuseppe & Colombo, A & Garofano, Luciano. (1995). Streptomyces peucetius daunorubicin biosynthesis gene, dnrF: Sequence and heterologous expression. Microbiology (Reading, England). 141 ( Pt 4). 1007-16. 10.1099/13500872-141-4-1007. [2] Hong, Young-Soo & Hwang, Cheol & Hong, Soon-Kwang & Kim, Young Ho & Lee, Jung Joon. (1994). Molecular cloning and characterization of the aklavinone 11-hydroxylase gene of Streptomyces peucetius subsp. caesius ATCC 27952. Journal of bacteriology. 176. 7096-101. 10.1128/jb.176.22.7096-7101.1994.

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