Part:BBa_K4821005
RdhANp_M132O_K12
This sequence codes for a modified RdhANp that has been docked to the ortho chlorine position in PCB 132 and codon-optimized for expression in E. coli K-12. This enzyme contains the following mutations: F294Y K295L E298L K310A P314Y M315L S423A S425V M426L N455I H457D V556I A563G. It was designed as an improvement on part BBa_K4821002, the codon-optimized wild-type RdhANp dehalogenase to possess higher catalytic activity against PCB 132.
Usage and Biology
This is a modified RdhANp with the potential to dechlorinate PCB132 at the ortho position for more efficient breakdown of the compound to acetyl CoA by the biphenyl degradation pathway (bph gene cluster). Mechanistically, Y426 in RdhANp protonates the chlorine located at 3.5A from the oxygen of Y426 and 5.0A from the cobalamin to catalyze a dechlorination reaction where the chlorine would coordinate with the Co on the cobalamin.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1628
Illegal BamHI site found at 312
Illegal BamHI site found at 419 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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