Coding

Part:BBa_K2317003

Designed by: Shan Wang   Group: iGEM17_Jilin_China   (2017-10-22)
Revision as of 12:08, 1 November 2017 by Eleven (Talk | contribs)

cphA-1

cphA-1 is a catechol 1,2-dioxygenase from Arthrobacter chlorophenolicus A6, and isresponsible for ring cleavage in aromatic compounds degrading.

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
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 214
    Illegal NgoMIV site found at 391
    Illegal NgoMIV site found at 757
  • 1000
    COMPATIBLE WITH RFC[1000]


Description

It was found that the cphA-1 enzyme could catalyze the degradation of catechol, 4-chlorocatechol, and 3-methylcatechol.


In our project, cphA-1 gene is cloned into expression vector pET28a and induced by IPTG. The optimized induction condition is 0.2mM IPTG, 180rpm in 16℃ for 24h. The enzyme is further purified by affinity chromatography.


CphA-1 activity was determined by measuring the increase of absorbance at 260 nm in 37℃, corresponding to the maximum absorbance of product muconic acid.

[edit]
Categories
//cds/enzyme
Parameters
None