Difference between revisions of "Part:BBa K4447001"
(→Biology and Usage) |
|||
| Line 3: | Line 3: | ||
<partinfo>BBa_K4447001 short</partinfo> | <partinfo>BBa_K4447001 short</partinfo> | ||
| − | Erythromycin C-12 hydroxylase coding sequence from Saccharopolyspora erythraea. The enzyme is responsible for the stereospecific hydroxylation of the macrolactone ring present in erythromycin D and erythromycin B. | + | Erythromycin C-12 hydroxylase coding sequence from <i>Saccharopolyspora erythraea</i>. The enzyme is responsible for the stereospecific hydroxylation of the macrolactone ring present in erythromycin D and erythromycin B. |
| Line 15: | Line 15: | ||
=Usage and Biology= | =Usage and Biology= | ||
| − | + | <div style="text-align:justify;"> | |
In the last few years, much attention has been drawn to emerging contaminants due to their severe effects on human health and the lack of information about them. Among them, erythromycin has risen as a potential threat in developing antimicrobial resistance, and being capable of detecting it in water bodies can lead to the generation of measurements capable of degrading it. | In the last few years, much attention has been drawn to emerging contaminants due to their severe effects on human health and the lack of information about them. Among them, erythromycin has risen as a potential threat in developing antimicrobial resistance, and being capable of detecting it in water bodies can lead to the generation of measurements capable of degrading it. | ||
| − | In our project, erythromycin C-12 hydroxylase is used as a detector for the presence of erythromycin by catalyzing the oxidation of two stereoisomers of erythromycin, erythromycin B and D, to erythromycin C, requiring NADPH as a reagent and, therefore, obtaining NADP+ as a reaction product. Consequently, it is possible to evaluate the presence of erythromycin through a coupled reaction employing a NADP+/NADPH colorimetric assay. | + | In our project, erythromycin C-12 hydroxylase is used as a detector for the presence of erythromycin by catalyzing the oxidation of two stereoisomers of erythromycin, erythromycin B and D, to erythromycin C, requiring NADPH as a reagent and, therefore, obtaining NADP+ as a reaction product. Consequently, it is possible to evaluate the presence of erythromycin through a coupled reaction employing a NADP+/NADPH colorimetric assay. The following image shows the complete reaction according to Stassi and collaborators (1993): |
| − | + | Erythromycin C-12 hydroxylase is a monomer with 397 amino acids in length. According to Savino <i>et al.<i> (2009), it binds one heme b(iron(II)-protoporphyrin IX) group per subunit as a cofactor. Lambalot <i>et al.</i> (1995) reported a Michaelis constant of 8 μM for erythromycin D, concluding that it shows a 1200-1900-fold preference for erythromycin D over the alternative substrate erythromycin B. This enzyme participates in various molecular and biological processes, ranging from macrolide biosynthetic processes to oxidoreductase reactions. | |
Revision as of 04:09, 27 September 2022
EryK coding sequence
Erythromycin C-12 hydroxylase coding sequence from Saccharopolyspora erythraea. The enzyme is responsible for the stereospecific hydroxylation of the macrolactone ring present in erythromycin D and erythromycin B.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1197
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Contents
Usage and Biology
In the last few years, much attention has been drawn to emerging contaminants due to their severe effects on human health and the lack of information about them. Among them, erythromycin has risen as a potential threat in developing antimicrobial resistance, and being capable of detecting it in water bodies can lead to the generation of measurements capable of degrading it.
In our project, erythromycin C-12 hydroxylase is used as a detector for the presence of erythromycin by catalyzing the oxidation of two stereoisomers of erythromycin, erythromycin B and D, to erythromycin C, requiring NADPH as a reagent and, therefore, obtaining NADP+ as a reaction product. Consequently, it is possible to evaluate the presence of erythromycin through a coupled reaction employing a NADP+/NADPH colorimetric assay. The following image shows the complete reaction according to Stassi and collaborators (1993):
Erythromycin C-12 hydroxylase is a monomer with 397 amino acids in length. According to Savino et al.<i> (2009), it binds one heme b(iron(II)-protoporphyrin IX) group per subunit as a cofactor. Lambalot <i>et al. (1995) reported a Michaelis constant of 8 μM for erythromycin D, concluding that it shows a 1200-1900-fold preference for erythromycin D over the alternative substrate erythromycin B. This enzyme participates in various molecular and biological processes, ranging from macrolide biosynthetic processes to oxidoreductase reactions.