Details zur Publikation |
| Kategorie | Textpublikation |
| Referenztyp | Zeitschriften |
| DOI | 10.1111/ejss.13225 |
Lizenz  |
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| Titel (primär) | An improved amplex red-based fluorometric assay of phenol oxidases and peroxidases activity: A case study on Haplic Chernozem |
| Autor | Khosrozadeh, S.; Dorodnikov, M.; Reitz, T.; Blagodatskaya, E. |
| Quelle | European Journal of Soil Science |
| Erscheinungsjahr | 2022 |
| Department | BOOEK |
| Band/Volume | 73 |
| Heft | 2 |
| Seite von | e13225 |
| Sprache | englisch |
| Topic | T5 Future Landscapes |
| Supplements | https://bsssjournals.onlinelibrary.wiley.com/action/downloadSupplement?doi=10.1111%2Fejss.13225&file=ejss13225-sup-0001-Supinfo.docx |
| Keywords | Amplex Red; Fluorometric assay; Michaelis-Menten kinetics; Peroxidase; Phenol oxidase activity |
| Abstract | The initial steps of complex organic matter degradation in soil comprise several oxidative processes catalyzed by phenol oxidases and peroxidases. Commonly, their activity is determined by colorimetric approaches based on a range of substrates, making comparison between studies difficult. To improve this shortcoming, we conducted a calibration procedure for assessing phenol oxidase and peroxidase activities in soils by a sensitive fluorometric assay. This assay is based on the colorless and non-fluorescent Amplex Red as a substrate, which forms a fluorescent product, resorufin, upon oxidation. Commercially available resorufin was used as a standard for calibration. We applied Michaelis-Menten kinetics to determine functional traits of phenol oxidases and peroxidases in a case study on Haplic Chernozem from different land-use and climate treatments of a field experiment. The kinetic parameters, limit of detection and limit of quantification demonstrated a high sensitivity of the fluorometric assay to differences caused by these treatments. For the total oxidative enzymes, the catalytic efficiency (Kcat, calculated as Vmax/Km) was 30% higher in the cropland compared to the pasture plot. However, compared to soil samples collected from ambient climatic conditions, Kcat of total oxidoreductases was 60% lower in cropland exposed to elevated air temperature and drought. The phenol oxidase activity was substantially higher than peroxidase activity in cropland land-use. However, higher Kcat of peroxidase than phenol oxidase resulted in 60% faster substrate turnover under both climatic conditions. In our case study, the Amplex Red-based assay enabled rapid and high-throughput analyses of phenol oxidase and peroxidase activity in soil. The sensitivity of this assay for soil enzymology applications needs further testing for a larger range of soils and environmental conditions. |
| dauerhafte UFZ-Verlinkung | https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=25797 |
| Khosrozadeh, S., Dorodnikov, M., Reitz, T., Blagodatskaya, E. (2022): An improved amplex red-based fluorometric assay of phenol oxidases and peroxidases activity: A case study on Haplic Chernozem Eur. J. Soil Sci. 73 (2), e13225 10.1111/ejss.13225 |
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