Part:BBa_K395602

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.

(→[http://2010.igem.org/Team:Tokyo_Tech/Project/Apple_Reporter2 more information])

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]

Functional Parameters: Austin_UTexas

Burden Imposed by this Part:

Burden Value: 20.3 ± 1.9%

Burden is the percent reduction in the growth rate of E. coli cells transformed with a plasmid containing this BioBrick (± values are 95% confidence limits). This part exhibited a significant burden. Users should be aware that BioBricks with a burden of >20-30% may be susceptible to mutating to become less functional or nonfunctional as an evolutionary consequence of this fitness cost. This risk increases as they used for more bacterial cell divisions or in larger cultures. Users should be especially careful when combining multiple burdensome parts, as plasmids with a total burden of >40% are expected to mutate so quickly that they become unclonable. Refer to any one of the BBa_K3174002 - BBa_K3174007 pages for more information on the methods and other conclusions from a large-scale measurement project conducted by the 2019 Austin_UTexas team.

This functional parameter was added by the 2020 Austin_UTexas team.