Team:NTU Taida/Circuit4

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NTU-Taida

Fragrance

  • Inspiration :

    GRASSY GREEN NOTES

    We want to turn fatty acids into volatile compounds that smells fresh and pleasant. First, we catalyze desaturation of Oleic acid(18:1 Δ9) into Linoleic acid(18:2 Δ9,12) by introducing a gene that encodesΔ12 Desaturase into E.coli (FAD6).

    Then we catalyze oxidation (LOX2) and hydrolysis (HPL1) of Linoleic acid(18:2 Δ9,12) to give a family of hexanal which smells fresh, generally known as “Grassy Green Notes”.

    ROSE FRAGRANCE

    Besides turning wasted fatty acids into fragrance, we also try to produce other fragrance from engineered E.coli. Our goal is to turn beta-carotene into rose fragrance by introducing a gene called CCD1 from Arabidopsis thaliana.

  • Testing Result :

    Function of Rose Fragrance Generator

    Method:

    Submit beta-ionone standard and extract of our rose fragrance generator to Gas Chromatography (GC) respectively

    Result:


    Beta-ionone standard
    We injected the beta-ionone standard, extracted by ethyl ehanol, into Gas chromotagraphy which existed an explicit curve at 27.5 minute.

    Extract of rose fragrance generator
    Then, we injected the sample which had been extracted by Ethyl ethanol also existed curves at around 27 minutes. However, the peak quality is not as good as standard.

    Beta-ionone standard


    Extract of rose fragrance generator

    Conclusion:

    Success in generating rose fragrance beta-ionone from our genetic engineered E.coli.

  • Background Knowledge :

    Green notes are fresh and lively and they are used to make a fragrance feel crisp and sharp. Green notes very often include green leaves, tea leaves, the essence of freshly cut grass and even some marine plants. Green notes are most commonly used in sporty fragrances and summertime editions of popular perfumes.

    Rose fragrance

    1. E.coli strain MG1063:
      • F plasmid with a transposon, Tn1000.
      • pSW140K with a beta-carotene synthesis operon.
      • Kanamycin resistance.

    2. E.coli strain HB101:
      • Without F plasmid.
      • pSW140K is stable in it. (Reference1)

    3. Carotenoid cleavage dioxygenase 1 (CCD1):
      • From Arabidopsis thaliana.
      • Turn beta-carotene into beta-ionone.
      • Full length CDS: 1617 bp

  • Reference :

    CCD1

    1. Mei-Hui Lin and Shih-Tung Liu (2008) J. Bacteriol 190(10):3681.
    2. To, Kin-Ying, et al. "Analysis of the gene cluster encoding carotenoid biosynthesis in Erwinia herbicola Eho13." Microbiology 140.2 (1994): 331-339.
    3. Cao, Hongbo, et al. "Comprehending crystalline β-carotene accumulation by comparing engineered cell models and the natural carotenoid-rich system of citrus." Journal of experimental botany (2012): ers115.
    4. http://cgsc.biology.yale.edu/Strain.php?ID=14667
    5. http://www.leffingwell.com/rose.htm
    FAD6

    LOX2

    HPL1

    Characterization of the yellow-pigment genes of Erwinia herbicola Molecular Microbiology (1991) 5(1). 217'-224

    Stabilization of pSW100 from Pantoea stewartii by the F Conjugation System JOURNAL OF BACTERIOLOGY, May 2008, p. 3681–3689 Vol. 190, No. 10 0021-9193/08/$08.000 doi:10.1128/JB.00846-07

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