Monitoring Lipase/Esterase Activity by Stopped Flow in a Sequential Injection Analysis System Using p-Nitrophenyl Butyrate

Sensors

Jorge Pliego⁴  Juan Carlos Mateos⁴  Jorge Rodriguez⁴  Francisco Valero³  Mireia Baeza¹  Ricardo Femat²  Rosa Camacho⁴  Georgina Sandoval⁴ 

[1] Departament de Química, Facultat de Ciències, Edifici C-Nord, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain; E-Mail:;Grupo de Biodinámica y Sistemas Alineales, División de Matemáticas Aplicadas, Instituto Potosino de Investigación Científica y Tecnológica. A.C. Camino a la Presa San José 2055, Lomas 4 Sección, C.P. 78216, San Luis Potosí S.L.P., Mexico; E-Mail:;Departament d'Enginyeria Química, Escola d'Enginyeria, Universitat Autònoma de Barcelona, 08193 Bellaterra, Barcelona, Spain; E-Mail:;Biotecnología Industrial, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco A.C., Avenida Normalistas 800, Colinas de la Normal. C.P. 44270, Guadalajara Jalisco, Mexico; E-Mails:

关键词: monitoring;    lipase/esterase activity;    sequential injection analysis;    stopped flow;    p-nitrophenyl esters;   

DOI  :  10.3390/s150202798

来源: mdpi

【 摘 要 】

Lipases and esterases are biocatalysts used at the laboratory and industrial level. To obtain the maximum yield in a bioprocess, it is important to measure key variables, such as enzymatic activity. The conventional method for monitoring hydrolytic activity is to take out a sample from the bioreactor to be analyzed off-line at the laboratory. The disadvantage of this approach is the long time required to recover the information from the process, hindering the possibility to develop control systems. New strategies to monitor lipase/esterase activity are necessary. In this context and in the first approach, we proposed a lab-made sequential injection analysis system to analyze off-line samples from shake flasks. Lipase/esterase activity was determined using p-nitrophenyl butyrate as the substrate. The sequential injection analysis allowed us to measure the hydrolytic activity from a sample without dilution in a linear range from 0.05–1.60 U/mL, with the capability to reach sample dilutions up to 1000 times, a sampling frequency of five samples/h, with a kinetic reaction of 5 min and a relative standard deviation of 8.75%. The results are promising to monitor lipase/esterase activity in real time, in which optimization and control strategies can be designed.

【 授权许可】

CC BY   
© 2015 by the authors; licensee MDPI, Basel, Switzerland.