Difference between revisions of "Part:BBa K5520011:Design"

 
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===Design Notes===
 
===Design Notes===
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We aimed to investigate the relationship between the structure and function of chitosanase and to produce chitooligosaccharides with a defined degree of polymerization. To achieve this, amino acid mutation at site 115 was engineered to enhance enzymatic activity and to generate more uniform products.
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We propose that the first step in the generation of chitosan from chitin, deacetylation of chitin to obtain chitosan, is important. The deacetylase encoded by CDA can catalyze the deacetylation of chitin to synthesize chitosan. We cloned the CDA gene into the PET28a vector to obtain the PET-28a-CDA recombinant plasmid, which was transformed into E. coli BL21 (DE3) to overexpress CDA.
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===Source===
 
===Source===
  
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Escherichia coli and Bacillus pumilus  
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Bacillus pumilus  
  
 
===References===
 
===References===

Revision as of 06:27, 30 September 2024


pT7-LacO-His- CsnBP115A


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

We propose that the first step in the generation of chitosan from chitin, deacetylation of chitin to obtain chitosan, is important. The deacetylase encoded by CDA can catalyze the deacetylation of chitin to synthesize chitosan. We cloned the CDA gene into the PET28a vector to obtain the PET-28a-CDA recombinant plasmid, which was transformed into E. coli BL21 (DE3) to overexpress CDA.


Source

Bacillus pumilus

References