Part:BBa_K5327014:Design

Myrosinase 1

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1563
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 421
  Illegal BsaI site found at 1030
  Illegal BsaI site found at 1403
  Illegal BsaI.rc site found at 1493
  

Design Notes

The design of the Myrosinase 1 gene is based on the coding sequence (CDS) from Arabidopsis thaliana and has been codon-optimized for Saccharomyces cerevisiae (S288C) to ensure its efficient expression. Myrosinase 1 is a key enzyme capable of degrading glucosinolates, and in this experiment, it primarily catalyzes the hydrolysis of Glucoraphanin, producing the target product Sulforaphane, which possesses anticancer, antibacterial, and antioxidant properties. The gene is expressed using the PGI1 promoter (PGI1pBBa_K5327017) and PYK1 terminator (PYK1tBBa_K5327018) to ensure high enzyme expression and mRNA stability in yeast. It is introduced into S. cerevisiae S288C via homologous recombination and screened and validated using auxotrophic selection. This design aims to enhance the activity of Myrosinase 1 in yeast, enabling efficient glucosinolate degradation and optimizing yeast as a metabolic engineering platform.

Plasmid

Fig 1. The plasmid expression of myrosinase 1

Source

Arabidopsis thaliana