Coding

Part:BBa_K2963004:Design

Designed by: Experiment group members of JNU-China   Group: iGEM19_JNU-China   (2019-09-07)
Revision as of 15:41, 18 October 2019 by GHX (Talk | contribs)


racE - a glutamate racemase from Bacillus sp.


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]


Design Notes

Determination of glutamate racemase activity: The cells ferment for 24 h, and the fermentation broth is centrifuged at 4°C. Collect the cells at 10000 rpm. Wash and centrifuge the cells using 0.85% physiological saline. Repeat the wash 3 times. The cells are suspended in p H 8.0, 0.1 M Tris-HCl buffer. Sonicate for 10 min in an ice bath. Centrifuge at 12000 rpm for 30 min, and the supernatant (crude enzyme solution) is taken for immediate determination of the relevant enzyme activity. Using 100 μL of crude enzyme solution reacts with 100 μL of substrate (0.5 g/L L-glutamic acid) at 30 ° C for 30 min and the reaction is terminated by boiling water bath. The content of D-glutamic acid in the reaction solution is determined by a high performance liquid phase method.

Source

Bacillus Genome.

References

1. Kimura, and K. "Roles and regulation of the glutamate racemase isogenes, racE and yrpC, in Bacillus subtilis." Microbiology 150.9(2004):2911-2920. 2. Ashiuchi, Makoto , et al. "Differences in effects on DNA gyrase activity between two glutamate racemases of Bacillus subtilis, the poly-γ-glutamate synthesis-linking Glr enzyme and the YrpC (MurI) isozyme." 223.2(2003):221-225.