Composite

Part:BBa_K4743034

Designed by: Jen Hsien, Liu   Group: iGEM23_PTSH-Taiwan   (2023-10-08)
Revision as of 02:21, 10 October 2023 by JenL (Talk | contribs)


Astaxanthin production set

The design of the astaxanthin biosynthesis pathway incorporates a range of crucial enzymes that facilitate the synthesis of astaxanthin starting from acetyl-CoA. This design adheres to the yeast's monocistronic system and employs the Adh1 constitutive promoter and Adh1 terminator to enable gene expression within yeast cells. Within this pathway, genes such as chyB, crtE, crtI, CrtYB, and bkt are responsible for encoding catalytic enzymes, including beta-carotene 3-hydroxylase, geranylgeranyl diphosphate synthase, phytoene dehydrogenase, Bifunctional lycopene cyclase/phytoene synthase, and β-carotene ketolase. These enzymes collectively drive the production of astaxanthin.

To aid in the selection process, a hygromycin-resistance gene, hph, has been incorporated into the genetic construct. This allows for the identification and maintenance of yeast cells that have successfully integrated the desired genetic material. It's worth noting that the gene tHMG-1 is naturally present in the Kluyveromyces Marxianus' genome.

The entire genetic construct is integrated into the pklac2 expression vector, which possesses the capacity for replication in E. coli and stable integration into the yeast genome, particularly in kluyveromyces lactis.

Overall metabolic pathway of Astaxanthin

Figure 1. The Matabolic pathway of Astaxanthin production.

Phenotyping

Figure 2. The phenotyping of our strain for the third generation.

Quantitative check

Figure 3. The HPLC test for the third generation.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 8176
    Illegal PstI site found at 8266
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 8176
    Illegal NheI site found at 627
    Illegal NheI site found at 915
    Illegal NheI site found at 1119
    Illegal NheI site found at 2274
    Illegal NheI site found at 9773
    Illegal NheI site found at 10061
    Illegal NheI site found at 10265
    Illegal PstI site found at 8266
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 8176
    Illegal BglII site found at 725
    Illegal BglII site found at 5611
    Illegal BglII site found at 9871
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 8176
    Illegal PstI site found at 8266
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 8176
    Illegal PstI site found at 8266
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 4377
    Illegal BsaI.rc site found at 12941


[edit]
Categories
Parameters
None