Coding

Part:BBa_K5327012

Designed by: Fangxian Chen   Group: iGEM24_BUCT   (2024-08-28)


S-alkyl-thiohydroximate lyase SUR1

Function:[1][2]

C-S lyase involved in glucosinolate biosynthesis. Converts S-(alkylacetohydroximoyl)-L-cysteine to thiohydroximate. Functions in auxin homeostasis. Probably required for glucosinolate activation in response to pathogens.

Usage and Biology

Genome localization:Chromosome: 2; NC_003071.7

Expression diagram:

Fig 1. The expression diagram of S-alkyl-thiohydroximate lyase SUR1

Corresponding enzyme structure:

Fig 2. The corresponding enzyme structure of S-alkyl-thiohydroximate lyase SUR1

The PCR result:

Fig 3. The PCR result of S-alkyl-thiohydroximate lyase SUR1

Subcellular localization:[3]

Located in the cytosol and mitochondrion of cells

Fig 4. The subcellular localization of S-alkyl-thiohydroximate lyase SUR1

Design Notes

The design of the S-alkyl-thiohydroximate lyase (SUR1) gene is based on the coding sequence (CDS) from Arabidopsis thaliana and has been codon-optimized for efficient expression in Saccharomyces cerevisiae (S288C). SUR1 plays a crucial role in glucosinolate biosynthesis, catalyzing the conversion of S-(alkylacetohydroximoyl)-L-cysteine to thiohydroximate, and may be involved in glucosinolate activation in response to pathogens.[4]To ensure high-level expression and mRNA stability, the design employs the HXT7 promoter(BBa_K4000003) and HXT7 terminator(BBa_K5327019). The optimized gene will be cloned into a vector, introduced into yeast S288C through homologous recombination, and validated using a knockout strain. This design aims to enhance the production efficiency of glucosinolates in yeast and further optimize yeast as a platform for metabolic engineering.

Plasmid

Fig 5. The plasmid expression of S-alkyl-thiohydroximate lyase SUR1

Source

Arabidopsis thaliana


References

  1. ↑ MIKKELSEN M D, NAUR P, HALKIER B A. Arabidopsis mutants in the C-S lyase of glucosinolate biosynthesis establish a critical role for indole-3-acetaldoxime in auxin homeostasis [J]. The Plant journal : for cell and molecular biology, 2004, 37(5): 770-7.
  2. ↑ DELARUE M, PRINSEN E, ONCKELEN H V, et al. Sur2 mutations of Arabidopsis thaliana define a new locus involved in the control of auxin homeostasis [J]. The Plant journal : for cell and molecular biology, 1998, 14(5): 603-11.
  3. ↑ https://www.uniprot.org/uniprotkb/Q9SIV0/entry
  4. ↑ MIKKELSEN M D, NAUR P, HALKIER B A. Arabidopsis mutants in the C-S lyase of glucosinolate biosynthesis establish a critical role for indole-3-acetaldoxime in auxin homeostasis [J]. The Plant journal : for cell and molecular biology, 2004, 37(5): 770-7.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 369
    Illegal XhoI site found at 529
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 526
    Illegal BsaI.rc site found at 892
    Illegal SapI site found at 165


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