Difference between revisions of "Part:BBa K2933001"
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<partinfo>BBa_K2933001 short</partinfo>
 
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NDM-23 is a type of subclass B metal beta-lactamases, which is derived from NDM-1 mutation. The beta lactamases of the NDM family can hydrolyze almost all available beta lactam antibiotics (except aztreonam) clinically, including the broad-spectrum antibiotic carbapenems. Because of the extensive substrate profile of this enzyme, the clinical strains carrying it become a great threat to human life and health.
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This part encodes a kind of beta lactamase called ndm-23.
  
 
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===Usage and Biology===
 
===Usage and Biology===
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NDM-23 is a type of subclass B metal beta-lactamases, which is derived from NDM-1 mutation. The beta lactamases of the NDM family can hydrolyze almost all available beta lactam antibiotics (except aztreonam) clinically, including the broad-spectrum antibiotic carbapenems. Because of the extensive substrate profile of this enzyme, the clinical strains carrying it become a great threat to human life and health.
  
  

Revision as of 13:31, 3 September 2019


subclass B1 metallo-beta-lactamase NDM-23, codon optimized in E. coli

This part encodes a kind of beta lactamase called ndm-23.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

NDM-23 is a type of subclass B metal beta-lactamases, which is derived from NDM-1 mutation. The beta lactamases of the NDM family can hydrolyze almost all available beta lactam antibiotics (except aztreonam) clinically, including the broad-spectrum antibiotic carbapenems. Because of the extensive substrate profile of this enzyme, the clinical strains carrying it become a great threat to human life and health.



References

Expression and purification

Pre-expression:
The bacteria were cultured in 5mL LB liquid medium with ampicillin in 37℃ overnight. After taking samples, we transfer them into 1L LB medium with ampicillin.

Massive expressing:
After 4 hours culturing in 37℃ in bottles, we used 500μM IPTG induced in 16℃ for 8-12h.

Purification of GST fusion proteins:

Anion exchange column:

Gel filtration chromatography:

Enzyme activity determination

Determination of enzyme concentration

Determination of the buffer composition

Determination of substrate concentration

Determination of the mikkelsen curve

Hydrolysis efficiency of different antibiotics

Inhibition of different inhibitors