Han Xiaoxun, Xie Yiming, Wu Qin, Wu Shuaibin
Hubei Key Laboratory of Pollutant Analysis & Reuse Technology, College of Chemistry and Chemical Engineering, Hubei Collaborative Innovation Center for Rare Metal Chemistry, Hubei Normal University, Cihu Road No. 11, Huangshi, China.
J Chromatogr Sci. 2019 Feb 1;57(2):116-121. doi: 10.1093/chromsci/bmy091.
In this work, two kinds of monolith-based immobilized enzyme microreactors (IMER) were compared in terms of monolith properties and digestion performance. Two monoliths were prepared involving poly (tetraethoxysiLane-co-3-aminopropyltriethoxysilane) (poly (TEOS-co-APTES)) monolith and poly (N-acryloxysuccinimide-co-acrylamide-co-ethylene dimethacrylate) (poly (NAS-co-AA-co-EDMA)) monolith for covalent trypsin immobilization. A higher chromatographic permeability constant 66.7 was obtained from the poly (TEOS-co-APTES) monolith in comparison with only 4.37 for poly (NAS-co-AA-co-EDMA) monolith, and the porous property of monolith measured by N2 adsorption/desorption and mercury intrusion porosimetry also indicated that poly (TEOS-co-APTES) monolith has more pores than poly (NAS-co-AA-co-EDMA) monolith. On the other hand, the digestion results demonstrated that the more efficient IMER was obtained by immobilizing trypsin on the poly (TEOS-co-APTES) monolith even if a less amount of immobilized trypsin determined by the Bradford method than poly (NAS-co-AA-co-EDMA) monolith. Therefore, the capacity of trypsin immobilization of monolith was not the exclusive factor to determine the digestion performance of IMER. Physical properties-penetrability and pore distribution of monolith showed a crucial effect on the digestion performance of IMER.
在本研究中,从整体柱性质和消化性能方面对两种整体柱固定化酶微反应器(IMER)进行了比较。制备了两种整体柱,分别是用于共价固定胰蛋白酶的聚(四乙氧基硅烷 - 共 - 3 - 氨丙基三乙氧基硅烷)(聚(TEOS - 共 - APTES))整体柱和聚(N - 丙烯酰氧基琥珀酰亚胺 - 共 - 丙烯酰胺 - 共 - 乙二醇二甲基丙烯酸酯)(聚(NAS - 共 - AA - 共 - EDMA))整体柱。聚(TEOS - 共 - APTES)整体柱获得了更高的色谱渗透常数66.7,而聚(NAS - 共 - AA - 共 - EDMA)整体柱仅为4.37,并且通过N2吸附/脱附和压汞法测量的整体柱多孔性质也表明聚(TEOS - 共 - APTES)整体柱比聚(NAS - 共 - AA - 共 - EDMA)整体柱具有更多的孔。另一方面,消化结果表明,即使通过Bradford法测定固定在聚(TEOS - 共 - APTES)整体柱上的胰蛋白酶量比聚(NAS - 共 - AA - 共 - EDMA)整体柱少,将胰蛋白酶固定在聚(TEOS - 共 - APTES)整体柱上仍能获得更高效的IMER。因此,整体柱固定胰蛋白酶的能力不是决定IMER消化性能的唯一因素。整体柱的物理性质——渗透性和孔分布对IMER的消化性能有至关重要的影响。