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Monitoring Lipase/Esterase Activity by Stopped Flow in a Sequential Injection Analysis System Using p-Nitrophenyl Butyrate
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 bioreacto...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4367334/ https://www.ncbi.nlm.nih.gov/pubmed/25633600 http://dx.doi.org/10.3390/s150202798 |
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author | Pliego, Jorge Mateos, Juan Carlos Rodriguez, Jorge Valero, Francisco Baeza, Mireia Femat, Ricardo Camacho, Rosa Sandoval, Georgina Herrera-López, Enrique J. |
author_facet | Pliego, Jorge Mateos, Juan Carlos Rodriguez, Jorge Valero, Francisco Baeza, Mireia Femat, Ricardo Camacho, Rosa Sandoval, Georgina Herrera-López, Enrique J. |
author_sort | Pliego, Jorge |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-4367334 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-43673342015-04-30 Monitoring Lipase/Esterase Activity by Stopped Flow in a Sequential Injection Analysis System Using p-Nitrophenyl Butyrate Pliego, Jorge Mateos, Juan Carlos Rodriguez, Jorge Valero, Francisco Baeza, Mireia Femat, Ricardo Camacho, Rosa Sandoval, Georgina Herrera-López, Enrique J. Sensors (Basel) Article 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. MDPI 2015-01-27 /pmc/articles/PMC4367334/ /pubmed/25633600 http://dx.doi.org/10.3390/s150202798 Text en © 2015 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Pliego, Jorge Mateos, Juan Carlos Rodriguez, Jorge Valero, Francisco Baeza, Mireia Femat, Ricardo Camacho, Rosa Sandoval, Georgina Herrera-López, Enrique J. Monitoring Lipase/Esterase Activity by Stopped Flow in a Sequential Injection Analysis System Using p-Nitrophenyl Butyrate |
title | Monitoring Lipase/Esterase Activity by Stopped Flow in a Sequential Injection Analysis System Using p-Nitrophenyl Butyrate |
title_full | Monitoring Lipase/Esterase Activity by Stopped Flow in a Sequential Injection Analysis System Using p-Nitrophenyl Butyrate |
title_fullStr | Monitoring Lipase/Esterase Activity by Stopped Flow in a Sequential Injection Analysis System Using p-Nitrophenyl Butyrate |
title_full_unstemmed | Monitoring Lipase/Esterase Activity by Stopped Flow in a Sequential Injection Analysis System Using p-Nitrophenyl Butyrate |
title_short | Monitoring Lipase/Esterase Activity by Stopped Flow in a Sequential Injection Analysis System Using p-Nitrophenyl Butyrate |
title_sort | monitoring lipase/esterase activity by stopped flow in a sequential injection analysis system using p-nitrophenyl butyrate |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4367334/ https://www.ncbi.nlm.nih.gov/pubmed/25633600 http://dx.doi.org/10.3390/s150202798 |
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