Composite

Part:BBa_K5520010:Design

Designed by: Ruyi Shi   Group: iGEM24_Ulink-SZ   (2024-09-29)


pT7-LacO-His- CsnBD78Y


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 844
    Illegal NotI site found at 804
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 813
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 153
    Illegal AgeI site found at 636
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Chitosanase catalyzes colloidal chitosan to generate chitooligosaccharides whose degree of polymerization still needs to be improved, and to obtain chitooligosaccharides products of a specific degree of polymerization, several methods have been developed to improve the distribution of DP in the final enzymatic hydrolysis product of the product, but it has been verified that interfering with substrate binding in the active site and altering the propensity to substrate binding are still the most effective strategies to improve the specificity of the enzyme product. Therefore, protein engineering of chitosanase gene is very important to explore the relationship between structure and function of chitosanase as well as to prepare products with specific degree of polymerization of chitooligosaccharides, so amino acid mutation of site 78 was designed in order to obtain enzyme of higher activity and more homogeneous products.


Source

Escherichia coli and Bacillus pumilus

References