The engineering and immobilization of penicillin G acylase onto thermo‐sensitive tri‐block copolymer system

• Published in: Polymers for advanced technologies 2019-01, Vol.30 (1), p.86-93
• Main Authors: Li, Ke, Liu, Xiao Ting, Zhang, Yun Fei, Liu, Donglei, Zhang, Xin Yu, Ma, Song Mei, Ruso, Juan M., Tang, Zhenghua, Chen, Zhen Bin, Liu, Zhen
• Format: Article
• Language: English
• Subjects: Antibiotics; Block copolymers; carrier; Catalysis; Enzyme activity; Enzymes; Immobilization; PDEA‐b‐PHEMA‐b‐PGMA; Penicillin; Penicillin G acylase; Polyhydroxyethyl methacrylate; tri‐block thermo‐sensitive copolymer
• ISSN: 1042-7147; 1099-1581
• DOI: 10.1002/pat.4446

In this work, the relationships between catalytic performances of penicillin G acylase (PGA) and the molar ratio of carrier, thermo‐sensitive tri‐block polymer, poly (N,N‐diethylacrylamide‐b‐ β‐hydroxyethyl methacrylate‐b‐glycidyl methacrylate) (PDEA‐b‐PHEMA‐b‐PGMA) were studied firstly, and result documented the optimal molar ratio was nDEA:nHEMA:nGMA = 100:47:24, which presented a suitable lower critical solution temperature (LCST) of 35°C and the activity retention ratio of 80.62% (±0.50%). Based on the suitable carrier, immobilization conditions were investigated and optimized. When pH of solution, concentration of PGA, immobilized time, and immobilization temperature were 8.0, 1/10 (m/v), 16 hours, and 36°C, respectively, enzyme loading capacity (L), enzyme activity (Ea), and activity retention ratio (Ar) of PGA arrived at the highest value of 21 223 U, 16 199 U/g, and 93.50% (±0.50%), respectively. Besides, the response rate (Rr) of immobilized PGA was the same as free PGA, the reusable stability (Rs) was 77.00% (±1.00%) after using for 11 times, which indicated that the carrier has better compatibility with L, Ar, Rs, and Rr.