Functionalized Mesoporous Silica with Very Large Pores for Cellulase Immobilization

Organo-functionalized FDU-12 type silicas exhibiting large pore sizes and ordered mesoporous structures were synthesized at low reaction (15 degrees C) and high hydrothermal (160 degrees C) temperatures via the co-condensation of tetraethoxysilane (TEOS) with a suite of organosilanes, i.e., 3-aminopropyltriethoxysilane (APTES), 3-mercaptopropyltrimethoxysilane (MPTMS), vinyltrimethoxysilane (VTMS), and phenyltrimethoxysilane (PTMS), in the presence of structure directing micelles formed using the surfactant pluronic F127 and the pore enlarging reagent trimethylbenzene (TMB). Small-angle X-ray scattering (SAXS) and transmission electron microscopy (TEM) confirmed that all synthesized materials possessed a face-centered cubic mesostructure (space group Fm (3) over barm), while nitrogen sorption analyses showed that the synthesized materials had extra large pores with cavity sizes of up to 25.4 nm and entrance sizes of up to 10.8 nm. X-ray photoelectron spectroscopy (XPS) and C-13 solid-state magic-angle spinning nuclear magnetic resonance (C-13-MAS NMR) measurements verified the incorporation of the different organosilanes into the silica network and more importantly on the inner and outer surfaces of the materials. As-obtained mesoporous silicas were tested in protein immobilization studies using bovine serum albumin and the cellulose-hydrolyzing enzyme cellulase, which in itself is a mixture of three large enzymes. Enzyme immobilization efficiency, activity, and stability varied significantly with organic functionality due to size exclusion effects at pore entries, electrostatic and hydrophobic interactions between the organo-functionalized surfaces and the enzymes, and conformational changes of the enzymes which can occur on some of the material surfaces. As a result, phenyl (PTMS)- and thiol (MPTMS)-functionalized FDU-12 mesoporous silicas had a very low adsorption capacity of proteins because of their small pore sizes. Amino (APTES)-functionalized FDU-12 mesoporous silica showed the highest adsorption amount of proteins yet the lowest activity of immobilized cellulase. Cellulase immobilization on vinyl (VTMS)-functionalized FDU-12 mesoporous silica appeared to be the most promising approach, since it occurred with high efficiency, maintained enzyme activity, and provided temporal enzyme stability.