Gazi University, Faculty of Arts and Sciences, Biochemical Processing and Biomaterial Research Laboratory, Teknik Okullar, 06500 Ankara, Turkey.
Bioresour Technol. 2011 Feb;102(4):3653-61. doi: 10.1016/j.biortech.2010.12.029. Epub 2010 Dec 13.
Fibrous poly(styrene-d-glycidylmethacrylate) (P(S-GMA)) brushes were grafted on poly(styrene-divinylbenzene) (P(S-DVB)) beads using surface initiated-atom transfer radical polymerization (SI-ATRP). Tetraethyldiethylenetriamine (TEDETA) ligand was incorporated on P(GMA) block. The multi-modal ligand attached beads were used for reversible immobilization of catalase. The influences of pH, ionic strength and initial catalase concentration on the immobilization capacities of the P(S-DVB)-g-P(S-GMA)-TEDETA beads have been investigated. Catalase adsorption capacity of P(S-DVB-g-P(S-GMA)-TEDETA beads was found to be 40.8 ± 1.7 mg/g beads at pH 6.5 (with an initial catalase concentration 1.0mg/mL). The K(m) value for immobilized catalase on the P(S-DVB-g-P(S-GMA)-TEDETA beads (0.43 ± 0.02 mM) was found about 1.7-fold higher than that of free enzyme (0.25 ± 0.03 mM). Optimum operational temperature and pH was increased upon immobilization. The same support was repeatedly used five times for immobilization of catalase after regeneration without significant loss in adsorption capacity or enzyme activity.
纤维状聚(苯乙烯-二缩水甘油甲基丙烯酸酯)(P(S-GMA))刷通过表面引发原子转移自由基聚合(SI-ATRP)接枝在聚苯乙烯-二乙烯基苯(P(S-DVB))珠上。四乙二胺四乙酸(TEDETA)配体被掺入 P(GMA)嵌段中。多模态配体附着的珠子被用于过氧化物酶的可逆固定化。研究了 pH 值、离子强度和初始过氧化物酶浓度对 P(S-DVB)-g-P(S-GMA)-TEDETA 珠固定化容量的影响。在 pH 6.5(初始过氧化物酶浓度为 1.0mg/mL)下,P(S-DVB-g-P(S-GMA)-TEDETA 珠的过氧化物酶吸附容量为 40.8±1.7mg/g 珠。在 P(S-DVB-g-P(S-GMA)-TEDETA 珠上固定化过氧化物酶的 K(m)值(0.43±0.02mM)比游离酶高约 1.7 倍(0.25±0.03mM)。固定化后最佳操作温度和 pH 值升高。在再生后,相同的载体被重复用于五次过氧化物酶的固定化,而吸附容量或酶活性没有明显损失。