Generator

Part:BBa_K274210

Designed by: Siming Ma   Group: iGEM09_Cambridge   (2009-09-19)

CrtEBIY under constitutive promoter

This Biobrick is created by putting enzyme cassette CrtEBIY (with individual rbs) of Part BBa_K274200 under constitutive promoter R0011.

Enzyme cassette CrtEBIY (with individual rbs) of Part BBa_K274200 converts colourless farnesyl pyrophosphate to orange beta-carotene (via intermediates geranylgeranyl pyroiphosphate, phytoene and lycopene).

Amount of beta-carotene produced can be measured by photospectrometer with absorbance at 455nm (beta-carotene extraction using acetone).

Datasheet for Part BBa_K274200, Part BBa_274210 and Part BBa_274220 in E. coli strain MG1655. You may also wish to refer to the "Experience" page.

Carotene datasheet1.JPG Carotene datasheet2.JPG

For PDF version of this Datasheet: File:Carotene.pdf


Further Characterization of BBa_K274210 in MG1655 and TOP10 e. coli has been performed by The 2010 SDU-Denmark iGEM team.

Reference

Hal Alper, et al. Construction of lycopene-overproducing E. coli strains by combining systematic and combinatorial gene knockout targets. Nature Biotechnology 23 (2005).

Nishizaki T, et al. Metabolic engineering of carotenoid biosynthesis in Escherichia coli by ordered gene assembly in Bacillus subtilis. Appl Environ Microbiol. 2007 Feb

Luke Z. Yuan, et al. Chromosomal promoter replacement of the isoprenoid pathway for enhancing carotenoid production in E. coli. Metabolic Engineering 8 (2006).

Luan Tao, et al. Isolation of chromosomal mutations that affect carotenoid production in Escherichia coli: mutations alter copy number of ColE1-type plasmids. FEMS Microbiology Letters 243 (2005)

von Lintig J, et al. Filling the gap in vitamin A research. Molecular identification of an enzyme cleaving beta-carotene to retinal. J Biol Chem. 2000 Apr 21;275(16).


Sequence and Features

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2037
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1573
    Illegal NgoMIV site found at 1703
    Illegal AgeI site found at 788
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//function/biosynthesis
//function/reporter/pigment
Parameters
None