Akademik Veri Yönetim Sistemi
The 6th International Enzyme and Bioprocess Days, Kocaeli, Türkiye, 27 - 29 Ağustos 2025, ss.58, (Özet Bildiri)
General Information and Purpose: Enzymatic biotinylation is widely used for protein labelling, but free
enzyme contamination may limit its efficiency and specificity in some condition. In this study, TurboID was
fused with the Magnetoreceptor protein (MagR) to create a recombinant fusion enzyme. Its magnetic
nanoparticle binding and biotinylation activity were evaluated. The aim was to develop a magnetically
controllable, low-contamination system and broaden the applications of enzymatic biotinylation
Materials and Methods. Gene sequences were sourced from NCBI GenBank and designed with flexible
linkers to allow proper fusion. These were cloned into expression plasmids and introduced into E. coli for
protein production. After purification, enzyme activity was assessed, and the proteins were then attached to
magnetic nanoparticles (Fe3O4-SiO2). Finally, their biotinylation activity was tested in cell culture
Findings: TurboID-MagR enzyme production was carried out with an MBP-tag. The enzyme was
successfully purified, activity asses performed by comparing TurboID and bound to magnetic nanoparticles
as designed, and cell surface biotinylation activity was successfully demonstrated in cell culture experiments.
Discussion: In this study, the enzyme was successfully produced, although a slight decrease in activity and
aggregation was observed during purification. Despite these challenges, the system worked successfully in
vitro settings, promising for broader use in biological applications.
Conclusion: TurboID-MagR system was successfully produced and remained active despite some
limitations. Its magnetic controllability and activity highlight its potential for broader, contamination-reduced
applications.