Preparasi dan Karakterisasi Nanobiokatalis Mikroreaktor dengan Polimer Monolitik Nanopori Enzim Termobilisasi

Abstract

Nanobiocatalyst microreactor column of an immobilized trypsin onto nanoporous monolithic polymer has been developed. In this work, a poly (glycidyl methacrylate-co-ethylene glycol dimethacrylate) (poly(GMA-co-EDMA)) monolih with total monomer %T 40 and cross-linker %C 25 was prepared as a nanoporous polymeric-support by in-situ copolymerization inside the microbore silicosteel tubing (1.0 mm i.d x 100 mm). This nanoporous monolithic-support was chemically modified with trypsin through the ring-opening reaction of epoxide group to form a nanobiocatalyst microreactor. In order to obtain reliable microreactor, some parameters such as immobilization time (amount of trypsin), and effect of glutaraldehyde activation on the trypsin-immobilized monolithic column were optimized. The pore size distribution, morphology and elemental composition of the monolith were studied by inverse size exclusion chromatography (ISEC) and scanning electron microscopy (SEM-EDX), respectively. While permeability of the monolith was obtained through measurement of its back pressure using HPLC pump. The monolith was predominated by mesoporous content (65.85%) while the flow-through pore was 28.20%. From SEM-EDX, it was found that the highest nitrogen content (12.40%), which indicates the amount of trypsin chemically attached on the monolitic support, was found in the nanobiocatalyst microreactor with immobilization time 4h without glutaraldehyde as activation agent. The optimized nanobiocatalyst microreactor column was successfully applied to a rapid and efficient digestion of protein samples in the range of second to minute.Keywords: nanoporous monolit, immobilized enzyme, nanobiocatalyst microreactor, protein digestion Pada penelitian ini telah dilakukan pengembangan pembuatan kolom nanobiokatalis mikroreaktor yang terimobilisasi oleh enzim tripsin pada polimer monolitik nanopori. Polimer monolitik nanopori dibuat secara insitu kopolimerisasi dalam kolom silicosteel (1.0 mm i.d x 100 mm) dari monomer GMA dan EDMA sebagai crosslinker serta penambahan porogen dengan komposisi 1-propanol/1,4-butanediol/air (7:4:1) dan inisiator radikal AIBN 1%. Proses polimerisasi berlangsung selama 12 jam pada suhu 60°C dengan persentase total monomer %T 40 dan persentase crosslinker %C 25. Monolit dimodifikasi secara kimia menggunakan metode imobilisasi enzim melalui reaksi pembukaan cincin *Penulis korespondensi

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