RESUMEN
Silica has been extracted from rice husks via a simple hydrothermal process and functionalized with triethoxy(octyl)silane -OCTES (Octyl-SiO2) and (3-aminopropyl)triethoxysilane - 3-APTES (Amino-SiO2), with the aim of using it as support to immobilize lipase from Thermomyces lanuginosus (TLL) via adsorption. The supports have been characterized by particle size distribution and elemental analyses, XRD, TGA, SEM, AFM and N2 physisorption so as to confirm their functionalization. Effect of pH, temperature, initial protein loading and contact time on the adsorption process has been systematically evaluated. Maximum immobilized protein loading of 12.3 ± 0.1 mg/g for Amino-SiO2 (5 mM buffer sodium acetate at pH 4.0, 25 °C and initial protein loading of 20 mg/g) and 21.9 ± 0.1 mg/g for Octyl-SiO2 (5 mM buffer sodium acetate at pH 5.0, 25 °C and initial protein loading of 30 mg/g) was observed. However, these biocatalysts presented similar catalytic activity in olive oil emulsion hydrolysis (between 630 and 645 U/g). TLL adsorption was a spontaneous process involving physisorption. Experimental data on Octyl-SiO2 and Amino-SiO2 adsorption were well-fitted to the Langmuir isotherm model. It was also investigated whether these biocatalysts could synthesize cetyl esters via esterification reaction. Thus, it was found that cetyl stearate synthesis required 100-110 min of reaction time to attain maximum conversion percentage (around 94%). Ester productivity of immobilized TLL on Amino-SiO2 was 1.3-3.1 times higher than Octyl-SiO2.
Asunto(s)
Adsorción , Enzimas Inmovilizadas/metabolismo , Eurotiales/enzimología , Lipasa/metabolismo , Concentración de Iones de Hidrógeno , Hidrólisis , Cinética , Lipasa/aislamiento & purificación , Aceite de Oliva/metabolismo , Oryza/química , Dióxido de Silicio/aislamiento & purificación , Dióxido de Silicio/metabolismo , TemperaturaRESUMEN
Ion-exchange supports have been prepared via sequential functionalization of silica-based materials with (3Glycidyloxypropyl)trimethoxysilane (GPTMS) (Epx-SiO2) and activation with glycine (Gly-Epx-SiO2) in order to immobilize lipase from Thermomyces lanuginosus (TLL) via adsorption. Rice husk silica (RHS) was selected as support with the aim of comparing its performance with commercial silica (Immobead S60S). Sequential functionalization/activation of SiO2-based supports has been confirmed by AFM, SEM and N2 adsorption-desorption analyses. Maximum TLL adsorption capacities of 14.8⯱â¯0.1â¯mg/g and 16.1⯱â¯0.6â¯mg/g using RHS and Immobead S60S as supports, respectively, have been reached. The Sips isotherm model has been used which was well fitted to experimental data on TLL adsorption. Catalytic activities of immobilized TLL were assayed by olive oil emulsion hydrolysis and butyl stearate synthesis via an esterification reaction. Hydrolytic activity of the biocatalyst prepared with a commercial support (357.6⯱â¯11.2â¯U/g) was slightly higher than that of Gly-Epx-SiO2 prepared with RHS (307.4⯱â¯7.2â¯U/g). On the other hand, both biocatalysts presented similar activity (around 90% conversion within 9-10â¯h of reaction) and reusability after 6 consecutive cycles of butyl stearate synthesis in batch systems.
Asunto(s)
Biocatálisis , Enzimas Inmovilizadas/metabolismo , Compuestos Epoxi/química , Eurotiales/enzimología , Glicina/química , Lipasa/metabolismo , Dióxido de Silicio/química , Enzimas Inmovilizadas/química , Esterificación , Concentración de Iones de Hidrógeno , Hidrólisis , Intercambio Iónico , Lipasa/química , Estearatos/químicaRESUMEN
In this work, mesoporous silica mobil composition of matter no. 41 (MCM-41) was synthesized by the sol-gel method. Two different surface modifications were made to transform this material into a very active adsorbent and catalyst support: (i) impregnation of iron nanoparticles and (ii) hydrophobization via chemical vapor deposition (CVD) with ethanol. The materials prepared with different iron contents, i.e., 2.5, 5, and 10 %, after hydrophobization, were characterized by several techniques. CHN analysis and Raman spectroscopy proved that approximately 15 % of carbon is deposited during CVD process mainly as organized carbonaceous structures. The specific surface area was determined by the BET method as up to 1080 m2 g-1, which explains the excellent results of the materials in the adsorption of model dyes methylene blue and indigo carmine. Mössbauer spectroscopy, thermogravimetric (TG)/DTG analysis, and transmission electron microscopy (TEM) images showed that the iron supported may be partially reduced during the CVD process to Fe2+ species, which are stabilized by the carbon coating. This iron species plays an important role in the oxidation of different contaminants, such as quinoline and methylene blue. The results obtained in the catalytic tests showed to be very promising.