Direct Electrochemical Generation of Catalytically Competent Oxyferryl Species of Classes I and P Dye Decolorizing Peroxidases
Abstract
:1. Introduction
2. Results
2.1. Generation and Spectroscopic Characterization of BsDyP and PpDyP High-Valent Catalytic Intermediates
2.2. Aerobic versus Anaerobic Redox Titrations of BsDyP and PpDyP
2.3. The Electrochemically Generated Oxyferryl Species Are Catalytic
3. Materials and Methods
3.1. Chemicals and Buffer Solutions
3.2. Construction, Overproduction and Purification of BsDyP and PpDyP
3.3. UV-Vis and Resonance Raman Experiments
3.4. Spectroelectrochemical Experiments
3.5. Steady State Kinetic Assays
3.6. Cyclic Voltammetry
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Enzyme | Mode | Resting State | Compound I | Compound II | Δνox |
---|---|---|---|---|---|
BsDyP | ν4 | 1373 | 1373 | 1377 | −4 |
ν28 | 1428 | 1428 | 1430 | −2 | |
ν3 | 1505 | 1505 | 1510 | −5 | |
ν19 | - | 1554 | 1553 | +1 | |
ν2 | 1561 | 1563 | 1564 | −1 | |
ν37 | 1579 | 1580 | 1582 | −2 | |
ν10 | - | 1600 | 1602 | −2 | |
1621 | 1619 | 1621 | −2 | ||
νC=C | 1634 | 1636 | 1640 | −4 | |
UV-vis bands | |||||
406, 505, 540, 632 | 406, 524, 565 | 418, 528, 556 | |||
PpDyP | ν4 | 1372 | 1374 | 1375 | −1 |
ν28 | 1428 | 1426 | 1430 | −4 | |
ν3 | 1503 | 1506 | 1508 | −2 | |
ν19 | 1557 | 1557 | 1553 | +3 | |
ν2 | 1567 | 1565 | 1567 | −2 | |
ν37 | 1581 | 1580 | 1581 | −1 | |
ν10 | 1621 | 1620 | 1621 | −1 | |
νC=C | 1631 | 1631 | 1637 | −6 | |
UV-vis bands | |||||
405, 506, 632 | 405, 530, 562, 644 | 419, 528, 557 |
pH | kcat (s−1) | KM (mM) | kcat/KM (s−1 M−1) | ||
---|---|---|---|---|---|
BsDyP | 1 H2O2 | 3.7 | 15 ± 1 | 0.420 ± 0.030 | 3.6 × 104 |
Electrochemical | 3.7 | 13 ± 1 | 0.141 ± 0.040 | 9.2 × 104 | |
7.6 | 18 ± 1 | 1.155 ± 0.100 | 1.6 × 104 | ||
8.5 | 1.1 ± 0.1 | 2.334 ± 0.060 | 4.7 × 102 | ||
PpDyP | 2 H2O2 | 4 | 17 ± 1 | 1.349 ± 0.115 | 1.3 × 104 |
Electrochemical | 3.7 | 4.2 ± 0.4 | 0.094 ± 0.016 | 4.5 × 104 | |
7.6 | 19 ± 2 | 1.343 ± 0.190 | 1.4 × 104 | ||
8.5 | 3.2 ± 0.2 | 0.070 ± 0.010 | 4.6 × 104 |
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Scocozza, M.F.; Martins, L.O.; Murgida, D.H. Direct Electrochemical Generation of Catalytically Competent Oxyferryl Species of Classes I and P Dye Decolorizing Peroxidases. Int. J. Mol. Sci. 2021, 22, 12532. https://doi.org/10.3390/ijms222212532
Scocozza MF, Martins LO, Murgida DH. Direct Electrochemical Generation of Catalytically Competent Oxyferryl Species of Classes I and P Dye Decolorizing Peroxidases. International Journal of Molecular Sciences. 2021; 22(22):12532. https://doi.org/10.3390/ijms222212532
Chicago/Turabian StyleScocozza, Magalí F., Lígia O. Martins, and Daniel H. Murgida. 2021. "Direct Electrochemical Generation of Catalytically Competent Oxyferryl Species of Classes I and P Dye Decolorizing Peroxidases" International Journal of Molecular Sciences 22, no. 22: 12532. https://doi.org/10.3390/ijms222212532