Difference between revisions of "Part:BBa K4447002"

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(→‎Usage and Biology)
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In our project, pentachlorophenol 4-monooxygenase (EC.1.14.13.50) is used as a detector for the presence of pentachlorophenol by catalyzing the dechlorination of pentachlorophenol to tetrachlorobenzoquinone, requiring NADPH as a reagent and, therefore, obtaining NADP+ as a reaction product. Consequently, it is possible to evaluate the presence of pentachlorophenol through a coupled reaction employing a NADP+/NADPH colorimetric assay. The following image shows the complete reaction according to Hlouchova <i>et al.</i> (2012):
 
In our project, pentachlorophenol 4-monooxygenase (EC.1.14.13.50) is used as a detector for the presence of pentachlorophenol by catalyzing the dechlorination of pentachlorophenol to tetrachlorobenzoquinone, requiring NADPH as a reagent and, therefore, obtaining NADP+ as a reaction product. Consequently, it is possible to evaluate the presence of pentachlorophenol through a coupled reaction employing a NADP+/NADPH colorimetric assay. The following image shows the complete reaction according to Hlouchova <i>et al.</i> (2012):
  
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Pentachlorophenol 4-monooxygenase is a dimeric protein that belongs to the family of flavin-dependent phenol hydroxylases. It has 539 amino acids in length and 60.1 kDa in weight (Cai & Xun, 2002). According to Hlouchova <i>et al.</i> (2012) reported a Michaelis constant of 1 mM for pentachlorophenol, concluding that this enzyme is not well evolved for turnover of this substrate. Nevertheless, this value is smaller than the one for 2,3,5,6-tetrachlorophenol, showing more preference for our desired substrate. Next, we present the three-dimensional structure of EryK generated by AlphaFold2 using MMSeqs2 (Mirdita et al., 2022). This structure is as follows:
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Pentachlorophenol 4-monooxygenase (Pcp) is a dimeric protein that belongs to the family of flavin-dependent phenol hydroxylases. It has 539 amino acids in length and 60.1 kDa in weight (Cai & Xun, 2002). According to Hlouchova <i>et al.</i> (2012) reported a Michaelis constant of 1 mM for pentachlorophenol, concluding that this enzyme is not well evolved for turnover of this substrate. Nevertheless, this value is smaller than the one for 2,3,5,6-tetrachlorophenol, showing more preference for our desired substrate. Next, we present the three-dimensional structure of Pcp generated by AlphaFold2 using MMSeqs2 (Mirdita et al., 2022). This structure is as follows:

Revision as of 16:13, 28 September 2022


Pcp coding sequence

Pentachlorophenol 4-monooxygenase coding sequence from Flavobacterium sp. This enzyme, a FAD binding and NADPH requiring oxygenase, catalyzes the oxygenolytic removal of the first chlorine from pentachlorophenol.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1651
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

In our project, pentachlorophenol 4-monooxygenase (EC.1.14.13.50) is used as a detector for the presence of pentachlorophenol by catalyzing the dechlorination of pentachlorophenol to tetrachlorobenzoquinone, requiring NADPH as a reagent and, therefore, obtaining NADP+ as a reaction product. Consequently, it is possible to evaluate the presence of pentachlorophenol through a coupled reaction employing a NADP+/NADPH colorimetric assay. The following image shows the complete reaction according to Hlouchova et al. (2012):

Pentachlorophenol 4-monooxygenase (Pcp) is a dimeric protein that belongs to the family of flavin-dependent phenol hydroxylases. It has 539 amino acids in length and 60.1 kDa in weight (Cai & Xun, 2002). According to Hlouchova et al. (2012) reported a Michaelis constant of 1 mM for pentachlorophenol, concluding that this enzyme is not well evolved for turnover of this substrate. Nevertheless, this value is smaller than the one for 2,3,5,6-tetrachlorophenol, showing more preference for our desired substrate. Next, we present the three-dimensional structure of Pcp generated by AlphaFold2 using MMSeqs2 (Mirdita et al., 2022). This structure is as follows: