Coding
ptxD B3-B4

Part:BBa_K2984013

Designed by: Sophia Schiebler   Group: iGEM19_Humboldt_Berlin   (2019-10-13)

Phosphite oxidoreductase (ptxD) B3-B4

This part is a part of the Chlamy-HUB-Collection. One restricting factor for cultivating microalgae is the high risk of contamination with bacteria, fungi or zooplankton. As our team anticipates in building a bioreactor it is essential to sterilize the media and all containers which raises the costs of production. Loera-Quezada et al. (2016) found a way to bypass this problem by integrating Pseudomonas stutzeri WM88 gene ptxD in C. reinhardtii. It encodes a phosphite oxidoreductase which is able to oxidize phosphite into phosphate using NAD+ as a cofactor (White & Metcalf, 2007). The integration of ptxD into the genome of C. reinhardtii enables the use of phosphite as a sole phosphorus source which wasn’t possible before (Loera-Quezada et al., 2015). Loera-Quezada et al. (2016) presented a new way of microalgae production without the need for sterile conditions or antibiotics to avoid contamination with biological pollutants as they can’t use phosphite as a phosphorus source. In the future this system could also be applied to open pond systems.

This part was designed to be used with the MoClo standard and has B3-B4 overhangs.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 967
  • 1000
    COMPATIBLE WITH RFC[1000]

Characterization

Plate_L0-ptxD_B3_B3_E.coli
Fig.1 - Successful transformation of L0-ptxD B3-B4 into E. coli.

References

  1. Loera‐Quezada, M. M., Leyva‐González, M. A., Velázquez‐Juárez, G., Sanchez‐Calderón, L., Do Nascimento, M., López‐Arredondo, D., & Herrera‐Estrella, L. (2016). A novel genetic engineering platform for the effective management of biological contaminants for the production of microalgae. Plant biotechnology journal, 14(10), 2066-2076.
  2. White, A. K., & Metcalf, W. W. (2007). Microbial metabolism of reduced phosphorus compounds. Annu. Rev. Microbiol., 61, 379-400.
  3. Loera-Quezada, M. M., Leyva-González, M. A., López-Arredondo, D., & Herrera-Estrella, L. (2015). Phosphite cannot be used as a phosphorus source but is non-toxic for microalgae. Plant Science, 231, 124-130.
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Categories
//cds
//cds/biosynthesis
Parameters
function
protein