Generator
MpAAT1

Part:BBa_K395602

Designed by: Toshitaka Matsubara   Group: iGEM10_Tokyo_Tech   (2010-10-04)

Apple fragrance generator

BBa_K395602 produce MpAAT1 enzyme that catalyzes production of 2-methylbutyl acetate and butyl acetate from 2-methylbutanol and butanol. 2-methylbutyl acetate and butyl acetate has a apple fragrance.


We designed apple fragrance expression device with MpAAT1[1]. Fig. 1 shows the outline of the device. MpAAT1 converts substrate and Acetyl CoA into apple fragrance molecules. Acetyl CoA exists originally in E.coli cell. MpAAT1 converts 2-methyl butanol and butanol into 2-methylbutyl acetate and butyl acetate respectively.


Fig. 1. Apple fragrance expression device.


We performed gas chromatography to confirm the production of the esters, and the results revealed that MpAAT1 successfully converted alcohol and Acetyl CoA into our target esters (Fig. 2).

Fig. 2. Gas chromatography analysis of apple fragrance expression device.
(1) MpAAT1 + BuOH, (2) No MpAAT1 + BuOAc, (3) No MpAAT1 + BuOH, (4) MpAAT1 + 2-MeBuOH, (5) No MpAAT1 + 2-MeBuOAc, (6) No MpAAT1 + 2-MeBuOH, (7) MpAAT1 - 2-MeBuOH.
This work is done by Toshitaka Matsubara.

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Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1765
    Illegal NotI site found at 1666
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 195
    Illegal BamHI site found at 1635
    Illegal XhoI site found at 1675
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 205
    Illegal AgeI site found at 1419
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None