Electrophoresis and biophysical characterization of sarcosine oxidase - Modification of magnetic nanoparticles

• Main Authors: Dagmar Uhlirova, Catia Damião, Martina Stankova, Michaela Vsetickova, Zuzana Tothova, Branislav Ruttkay-Nedecky, Marta Kepinska, Josef Ruzicka, Halina Milnerowicz, Warawan Eiamphungporn, Rene Kizek
• Other Authors: Universidade de Lisboa
• Format: Conference or Workshop Item
• Published: 2020
• Subjects: Biochemistry, Genetics and Molecular Biology; Engineering; Materials Science
• Online Access: https://repository.li.mahidol.ac.th/handle/123456789/60418
• Institution: Mahidol University

© NANOCON 2019.All right reserved. Sarcosine oxidase (SOX) is a flavoprotein and cleaves sarcosine to form hydrogen peroxide, glycine and formaldehyde. 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.