Generator

Part:BBa_K1602017

Designed by: Leon Kraus, Jonathan Pletzer-Zelgert, Jonas Sindlinger, Carmen Sakas Gandullo, Sebastian Holzner, Elena Tonchevska, Mirjam Haun   Group: iGEM15_TU_Darmstadt   (2015-09-16)
Revision as of 22:52, 18 September 2015 by Henklem (Talk | contribs)

D-xylonic acid producing operon

D-Xylose is a monosaccharide belonging to the aldopentose family. It was recently shown that the D-xylose dehydrogenase xylB from Caulobacter crescentus can convert D-xylose to D-xylonolactone. This can react spontaneously or through the catalysation of xylC to D-xylonic acid.

In E. coli D-xylonic acid can further be metabolized to ethyleneglycol by the enzymes yjhG (BBa_K1602012), yagE (BBa_K1602011) and yqhD (BBa_K1602013) which are already present in this host.(1)


https://static.igem.org/mediawiki/parts/f/f1/TU_Darmstadt_EG_XylB-xylC.png

  • Figure 1 Sheme of the reactions catalyzed by xylB and xylC. The xylC reaction can also happen spontaneously but in a much lower speed. (2)


  • Usage

    This part is a composite of two coding genes with strong RBS (BBa_B0034). The transcription is controlled by a T7 promotor (BBa_I719005).

    We used this operon to investigate possible production of ethylene glycol in E. coli.


    Figure 2


    Results

    Figure 3 Scan of the PAGE containing from left to right a marker (M; Protein Marker III AppliChem), the positive sample (1) and a negative control (2). The picture was cropped and edited for clarification purposes.
    Figure 4 Plot of the gel lanes based on contrast analyses - created with ImageJ



    Sequence and Features


    Assembly Compatibility:
    • 10
      COMPATIBLE WITH RFC[10]
    • 12
      COMPATIBLE WITH RFC[12]
    • 21
      COMPATIBLE WITH RFC[21]
    • 23
      COMPATIBLE WITH RFC[23]
    • 25
      COMPATIBLE WITH RFC[25]
    • 1000
      COMPATIBLE WITH RFC[1000]


    References

    1. Liu H, Ramos KR, Valdehuesa KN, Nisola GM, Lee WK, Chung WJ. Biosynthesis of ethylene glycol in Escherichia coli. Appl Microbiol Biotechnol. 2013;97(8):3409-17.

    2. Toivari MH, Nygard Y, Penttila M, Ruohonen L, Wiebe MG. Microbial D-xylonate production. Appl Microbiol Biotechnol. 2012;96(1):1-8.

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