Optimization of enzymatic saccharification of wheat straw in a micro-scale system by reponse surface methodology
Optimización de la sacarificación enzimática de paja de trigo en microescala a través de la metodología de superficie de respuesta
C. Molina1*, A. Sánchez2, A. Serafín-Muñoz3 y J. Folch-Mallol4
1 Universidad de Guanajuato-Guanajuato, Depto. de Ingeniería Química, Noria Alta s/n, 36050 Guanajuato, Gto., México. * Corresponding author. E-mail: carlosmolinaS@hotmail.com.
2 Centro de Investigación y de Estudios Avanzados del IPN, Unidad de Ingeniería Avanzada. Av. del Bosque 1145, Colonia el Bajío, Zapopan, 45019, Jalisco, México.
]]> 3 Universidad de Guanajuato-Guanajuato, Depto. de Ingeniería Ambiental, Av. Juárez 77, Zona Centro, 36000, Guanajuato, Gto., México.4 Universidad Autónoma del Estado de Morelos, Centro de Investigación en Biotecnología, Cuernavaca, Morelos, México.
Received October 31, 2013.
Accepted July 2, 2014
Abstract
This paper studies the combined effects of temperature, pH and enzyme-substrate ratio (E/SR) on hydrolysis yield and specific reaction rate (SRV) in a microscale iystem in order to maximize enzymatic hydrolysis of pretreated wheat straw (WS). The WS was pretreated by alkaline -peroxide. Enzymatic complex Accellerase 1500TM was used for hydrolysis assays. Using response surface methodology, optimal parameter values were determined. A complete enzymatic kinetic of the hydrolysis reaction was obtained in 10 h. The optimal value of reducing sugars concentration (RSc), given by the model, was 5.97 mg/mL and the corresponding yield was 61.73%. The maximum yield for the WS hydrolysis was 61.73% and was achieved at a temperature of 52.0°C, pH 4.6, and a E/SR of 2.1 mL of Accellerase 1500TM/g of cellulose. The SRV was 4.80 U/mg and was obtained with the following conditions: pH 5.0, temperature of 48.5°C and an E/SR of 0.19 mL/g. A quadratic polynomial equation for predicting the hydrolysis yield was developed. The confirmation experiment showed a final value for RSc of 5.98 ± 0.81 mg/mL. This result indicates a % error of 0.33. The experimental results were in good agreement with predicted value.
Keywords: enzymatic saccharification, wheat straw, response surface methodology, microscale system.
]]> Resumen
El presente trabajo estudia la hidrólisis de paja de trigo utilizando un sistema de microreacción. El Método de Superficie de Respuesta se utilizó para estudiar los efectos combinados de la temperatura, el pH y la relación enzima-sustrato (RE/S) sobre la hidrólisis enzimática y la velocidad específica de reacción VER. El sustrato fue paja de trigo pretratada de forma alcalino-oxidativa. El extracto enzimático utilizado fue Accellerase 1500TM. El tiempo de obtención de una cinética de la hidrólisis enzimática completa fue de 10 h. El valor óptimo de la concentración de azúcares reductores (CAR) arrojado por el modelo fue de 5.97 mg/mL y el rendimiento correspondiente fue de 61.73%. Estos valores fueron obtenidos con una temperatura de 52.0°C, pH 4.6 y una RE/S de 2.1 mL de Accellerase 1500/g de celulosa. La VER óptima fue de 4.80 U/mg y fue obtenida con una temperatura de 48.5°C, pH 5.0 y una RE/S de 0.19 mL/g. Se realizó un ensayo de confirmación en el que el valor predicho de CAR por el modelo fue de 5.96 mg/mL y el valor obtenido experimentalmente fue de 5.98 ± 0.81 mg/mL, indicando un error de 0.33%.
Palabras clave: hidrólisis enzimática, paja de trigo, metodología de superficie de respuesta, micro-escala, accellerase 1500TM.
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Acknowledgements
C.M. acknowledges financial support from CONACYT and the University of Guanajuato in the form of PhD scholarships. Financial support from project SENER 150001 is also acknowledged.
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