Coding

Part:BBa_K2286008

Designed by: Demin Xu   Group: iGEM17_UESTC-China   (2017-10-20)
Revision as of 14:08, 1 November 2017 by Tedemin (Talk | contribs) (→‎Activity toward 2，3-DCP)


HheC- P84A-W249P: halohydrin dehalogenase site-directed mutant

This is the site-directed mutant of halohydrin dehalogenase. Based on the excellent mutant HheC-W249P, We change the alanine to proline in the 84 site. And this sequence is codon-optimized for E.coli.

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 718
  • 1000
    COMPATIBLE WITH RFC[1000]

Characterization

This part is an improvement of BBa_K1199044 (W249P) in the activity of 2,3-DCP. We performed a superimposed mutation on the basis of W249P and obtained a mutant P84A-W249P with a higher activity than W249P.

Activity towards 2，3-DCP

In order to test the HheC catalytic activity towards 2,3-DCP, chloride ion method was applied by us. The results show that the catalytic activity of P84A-W249P to 2,3-DCP is 1.15 times than that of W249P.

Figure 1. The catalytic activity of mutants on 2,3-DCP. Data were measured in 50mM Tris-H2SO4 at pH8.5 and 37℃.




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