ELECTROPHORESIS AND BIOPHYSICAL CHARACTERIZATION OF SARCOSINE OXIDASE - MODIFICATION OF MAGNETIC NANOPARTICLES

1 UHLIROVA Dagmar
Co-authors:
2 DAMIÃO Catia 1 STANKOVA Martina 1 VSETICKOVA Michaela 1 TOTHOVA Zuzana 3 RUTTKAY-NEDECKY Branislav 4 KEPINSKA Marta 1 RUZICKA Josef 4 MILNEROWICZ Halina 5 EIAMPHUNGPORN Warawan 1,3,4 KIZEK Rene
Institutions:
1 Department of Research and Development, Prevention Medicals s.r.o., Tovární 342, 742 13; Studénka-Butovice, Czech Republic, uhlirova@preventionmedicals.cz
2 Faculty of Pharmacy University of Lisboa, Avenida Professor Gama Pinto, 1649-003, Lisboa, Portugal, catiassdamiao@gmail.com
3 Department of Human Pharmacology and Toxicology, University of Veterinary and Pharmaceutical Sciences Brno, Palackého třída 1946/1, 612 42 Brno, Czech Republic, kizek@sci.muni.cz
4 Department of Biomedical and Environmental Analyses, Faculty of Pharmacy with Division of Laboratory Diagnostics, Wroclaw Medical University, Borowska 211, 50-556 Wroclaw, Poland, zalewska.m@gmail.com
5 Department of Clinical Microbiology and Applied Technology, Faculty of Medical Technology, Mahidol University, 999 Phutthamonthon 4 Road, Salaya, Nakhon Pathom 73170, Thailand, warawan.eia@mahidol.ac.th
Conference:
11th International Conference on Nanomaterials - Research & Application, Hotel Voronez I, Brno, Czech Republic, EU, October 16th - 18th 2019
Proceedings:
Proceedings 11th International Conference on Nanomaterials - Research & Application
Pages:
580-585
ISBN:
978-80-87294-95-6
ISSN:
2694-930X
Published:
1st April 2020
Proceedings of the conference have been sent to Web of Science and Scopus for evaluation and potential indexing.
Metrics:
49 views / 15 downloads
Abstract

Sarcosine oxidase (SOX) is a flavoprotein and cleaves sarcosine to form hydrogen peroxide, glycine andformaldehyde. Sarcosine is commonly found in muscle, tissues, toothpaste and food supplements. Increased amounts have been found in patients with prostate cancer. SOX is an enzyme suitable for the enzymatic determination of sarcosine. Biotechnological applications require increased stability of used enzymes. In our study, we focused on temperature (5, 15, 20, 30, 35, 40, 50 and 60 °C) and pH-dependent changes (pH 5,5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10 and 10.5) in SOX activity. We used native polyacrylamide gel electrophoresis (Native-PAGE) to monitor changes in SOX. Native-PAGE can be sensitive to any process that changes either the charge or conformation of the protein, so they are excellent for detecting various chemical degradations, aggregations, binding changes, acidic, alkaline or denaturing in general. In addition, SOX was bound to the surface of magnetic nanoparticles (SPIONs). When SOX was bound to SPIONs and lyophilized,SOX activity remained unchanged (100 %) after 6 months.

Keywords: Sarcosine oxidase; sarcosine; enzymatic colorimetric assay; thermostability; Native-PAGE
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