Difference between revisions of "Part:BBa K2933001"
(→Enzymatic determination) |
(→Enzymatic determination) |
||
| Line 42: | Line 42: | ||
Gel filtration chromatography:<br> | Gel filtration chromatography:<br> | ||
| − | === | + | ===Enzyme activity determination=== |
====Determination of enzyme concentration==== | ====Determination of enzyme concentration==== | ||
Revision as of 13:16, 3 September 2019
subclass B1 metallo-beta-lactamase NDM-23, codon optimized in E. coli
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.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
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: