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

Abstract

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.

Design and synthesis of thermosensitive block resin as carrier for immobilization of penicillin G acylase

“…, which confirmed to the experimental result of Li et al (2019Li et al ( , 2018. Herein, in order to make the catalytic conversion temperature of immobilized PGA fallen into the temperature range of the sol state of thermosensitive block resin, a series of resins were prepared by employing poly (b -hydroxyethyl methacrylate) (PHEMA) as hydrophilic blocks and poly (glycidyl methacrylate) (PGMA) as hydrophobic blocks, and the LCST of which could be continuously adjusted by changing the segment length of hydrophilic blocks and hydrophobic blocks.…”

Immobilized penicillin G acylase with enhanced activity and stability using glutaraldehyde‐modified polydopamine‐coated Fe3O4 nanoparticles

“…With the elevation of temperature, the amounts of activated reactants increased; the quantities of 6‐APA increased relatively. However, the most active temperature range of PGA was 35–40°C 47 . When temperature was higher than 35–40°C, it distorted the conformation of hydrophobic pocket, resulted in the decrease of EA and EAR .…”

Thermoresponsive Poly(N,N-diethylacrylamide-co-glycidyl methacrylate) Copolymers and Its Catalytically Active α-Chymotrypsin Bioconjugate with Enhanced Enzyme Stability

“…It is interesting to note that due to the better biocompatibility of the LCST-type thermoresponsive poly( N , N -diethylacrylamide) (PDEAAm) than that of PNIPAAm [ 65 ], investigations in relation to the responsive and biocompatible behavior of PDEAAm, its derivatives, and gels have gained increased attention only in recent years (see, e.g., Refs. [ 66 , 67 , 68 , 69 , 70 , 71 , 72 , 73 , 74 , 75 , 76 , 77 , 78 , 79 , 80 , 81 , 82 , 83 , 84 , 85 , 86 , 87 , 88 , 89 , 90 , 91 , 92 ] and references therein). It should also be mentioned that aqueous PDEAAm solutions possess similar critical solution temperatures (CST) [ 72 , 93 , 94 , 95 , 96 ] in the range of ~25–40 °C than that of PNIPAAm [ 46 , 55 ].…”

“…It should also be mentioned that aqueous PDEAAm solutions possess similar critical solution temperatures (CST) [ 72 , 93 , 94 , 95 , 96 ] in the range of ~25–40 °C than that of PNIPAAm [ 46 , 55 ]. On the other hand, although some block copolymers with PDEAAm and poly(glycidyl methacrylate) (PGMA) segments have been prepared and studied [ 91 , 92 ], random copolymers of DEAAm with GMA, which provides reactive pendant epoxy functionalities for the thermoresponsive PDEAAm, and its utilization for polymer-based protein engineering have not been reported so far according to the best of our knowledge.…”