Immobilization of ficin on aminopropyl silica: Effects of activating agent on its stability and on its catalytic efficiency in dipeptide synthesis
Process Biochemistry, cilt.168, ss.174-187, 2026 (SCI-Expanded, Scopus)
- Yayın Türü: Makale / Tam Makale
- Cilt numarası: 168
- Basım Tarihi: 2026
- Doi Numarası: 10.1016/j.procbio.2026.06.015
- Dergi Adı: Process Biochemistry
- Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, BIOSIS, Chemical Abstracts Core, Compendex, INSPEC, Academic Search Ultimate (EBSCO), Engineering Source (EBSCO)
- Sayfa Sayıları: ss.174-187
- Anahtar Kelimeler: Antibacterial dipeptide, Dipeptide synthesis, Ficin, Genipin, Glutaraldehyde, Immobilization, Kinetically controlled synthesis
- Çukurova Üniversitesi Adresli: Evet
Özet
Ficin was immobilized on 3-aminopropyl-functionalized silica (Si-NH2) activated with genipin (Si-Genipin@ficin) or glutaraldehyde (Si-Glu@ficin). Immobilization pH and temperature, initial protein loading, and immobilization time were systematically optimized to achieve the maximum enzyme activity recovery. Immobilization was confirmed by FT-IR, SEM and SEM-EDS analyses. The optimum pH and temperature were determined to be 7.5 and 65°C for all ficin preparations. Thermal stability of Si-Genipin@ficin and Si-Glu@ficin increased approximately 5-fold compared to the free enzyme at 65°C. The free ficin retained only 5% of its initial activity after 30 days of storage at 25°C, whereas Si-Genipin@ficin, and Si-Glu@ficin retained 65% and 70% of their initial activities, respectively, under the same conditions. After 15 consecutive re-uses in hydrolysis reactions at pH 7.5 and 65°C, the residual activities of Si-Genipin@ficin, and Si-Glu@ficin were found to be 44% and 53%, respectively. Si-Genipin@ficin and Si-Glu@ficin were also tested for the synthesis of Z-isoleucyl glutamine (Z-Ile-Gln) using Z- L -isoleucine 4-nitrophenyl ester as acyl donor and L -Glutamine as nucleophile and results showed that at least 88% yield was obtained after 12 h reaction at pH 9.5 and 45°C. After five reuses in Z-Ile-Gn synthesis under these conditions, Si-Genipin@ficin, and Si-Glu@ficin retained 83% and 66% of their initial activities, respectively. This study demonstrates, for the first time, the comparative effectiveness of genipin- and glutaraldehyde-activated Si-NH2 supports for developing stable and reusable ficin biocatalysts with high efficiency in antibacterial dipeptide synthesis, highlighting their potential for industrial peptide production.