Sürdürülebilir Kalkınma Amaçları
Yayınlar
- MakaleYalnız künye2026Covalent Immobilization of Glutathione-S-Transferase onto Magnetic Iron Nanoparticles via Epichlorohydrin Spacer Arm
Kemal Caglar, M.; Gulesci, Nuri; Bilgin, Ramazan; Yucebilgic, Guzide
This study examined the synthesis of Fe₃O₄ magnetic nanoparticles (MNPs) and the covalent immobilization of glutathione-S-transferase (GSTs) onto these nanoparticles using an epichlorohydrin (ECH) spacer arm, as well as the optimal reaction conditions for both free and immobilized enzymes and the reusability of the immobilized enzyme. The optimal pH values for free and immobilized enzymes were established as 7.0 and 6.0, respectively. It was found that the ideal temperature for both free and immobilized enzymes was 37 and 40°C, respectively. Under the optimum conditions, the values of V_max for glutathione (GSH) and 1-chloro-2,4-dinitrobenzene (CDNB) substrates of free GSTs enzyme were measured as 204.8 U/mg prot. and 194.3 U/mg prot., respectively. The immobilized GSTs enzyme's CDNB and GSH substrates had V_max values of 33.29 U/mg prot. and 33.08 U/mg prot., respectively. The free GSTs enzyme's CDNB and GSH substrates have K_m values of 0.201 mM and 0.1873 mM, respectively. GSH and CDNB have K_m values of 0.3042 mM and 0.2523 mM, respectively. At various temperatures, we contrasted the thermal stability of free and immobilized GSTs. The immobilized GSTs enzyme retained 50% of its activity after 20 reuses. After 30 days of storage at 25°C and 4°C, it was discovered that 18% and 32% of the free GSTs enzyme activity were preserved, and 30 days of storage at 25°C and 4°C resulted in 20% and 41% preserved enzyme activity for the immobilized enzyme, respectively.
- MakaleAçık Erişim2026Layered Polymeric Carbon Nitride as a Green Support for Cellulase Immobilization: Improved Stability, Activity, and Reusability
Gulesci, Nuri; Altan, Orhan; Toprak, Ali; Yalçın, M. Serkan; Bilgin, Ramazan; Yildirim, Deniz
Polymeric carbon nitride (PCN) has attracted considerable attention in recent years due to its layered structure and donor groups such as terminal amines and triazine units. In this study, PCN was synthesized from melamine and employed for the first time as a support material for the immobilization of cellulase, a key enzyme used in numerous industrial applications. Transmission electron microscopy (TEM) analysis revealed that ranged from 219 to 375 nm in size, thereby confirming their nanoscale layered morphology. Cellulase from Aspergillus sp. was immobilized onto PCN using three methods. In the first approach, cellulase was adsorbed onto PCN to form PCN@cellulase. In the second method, PCN@cellulase was cross-linked with glutaraldehyde (PCN@cellulase/Glu). Finally, the primary amino group of PCN was modified with glutaraldehyde, and the cellulase was immobilized onto this support via covalent attachment (PCN/Glu@cellulase). Each cellulase preparation was assessed for its optimal pH, temperature conditions, thermal stability, and enzyme kinetics. The optimal pH was 5.5 for all cellulase preparations, while the optimal temperature was 45 °C for free cellulase and 55 °C for all immobilized cellulase preparations. Thermal stability of PCN@cellulase, PCN@cellulase/Glu, and PCN/Glu@cellulase increased by 2.1-, 2.7-, and 3.7-fold, respectively, compared to the free cellulase. PCN/Glu@cellulase showed a 1.4-fold higher catalytic efficiency than the free cellulase and retained 80% of its initial activity after 10 reuses. This work highlights PCN as a green, metal-free, and easy-to-prepare support for enzyme immobilization, thereby opening new possibilities for sustainable bioprocessing applications.