Glutaraldehyde cross-linking of immobilized thermophilic esterase on hydrophobic macroporous resin for application in poly(ε-caprolactone) synthesis

• Published in: Molecules (Basel, Switzerland) Vol. 19; no. 7; pp. 9838 - 9849
• Main Authors: Wang, Min, Shi, Hui, Wu, Di, Han, Haobo, Zhang, Jianxu, Xing, Zhen, Wang, Shuang, Li, Quanshun
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 08.07.2014; MDPI
• Subjects: Adsorption; cross linking; Cross-Linking Reagents - chemistry; Enzyme Stability; Enzymes; Enzymes, Immobilized - chemistry; Esterases - chemistry; Glutaral - chemistry; immobilized enzyme; Kinetics; Molecular weight; Polyesters; Polyesters - chemical synthesis; Polymerization; Proteins; Resins; ring-opening polymerization; Temperature; thermophilic esterase; ε-caprolactone; China
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules19079838

The immobilized thermophilic esterase from Archaeoglobus fulgidus was successfully constructed through the glutaraldehyde-mediated covalent coupling after its physical adsorption on a hydrophobic macroporous resin, Sepabeads EC-OD. Through 0.05% glutaraldehyde treatment, the prevention of enzyme leaching and the maintenance of catalytic activity could be simultaneously realized. Using the enzymatic ring-opening polymerization of ε-caprolactone as a model, effects of organic solvents and reaction temperature on the monomer conversion and product molecular weight were systematically investigated. After the optimization of reaction conditions, products were obtained with 100% monomer conversion and Mn values lower than 1010 g/mol. Furthermore, the cross‑linked immobilized thermophilic esterase exhibited an excellent operational stability, with monomer conversion values exceeding 90% over the course of 12 batch reactions, still more than 80% after 16 batch reactions.