Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Pliego, Jorge, Mateos, Juan Carlos, Rodriguez, Jorge, Valero, Francisco, Baeza, Mireia, Femat, Ricardo, Camacho, Rosa, Sandoval, Georgina, Herrera-López, Enrique J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2015
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
_version_ 1782362516863582208
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
work_keys_str_mv AT pliegojorge monitoringlipaseesteraseactivitybystoppedflowinasequentialinjectionanalysissystemusingpnitrophenylbutyrate
AT mateosjuancarlos monitoringlipaseesteraseactivitybystoppedflowinasequentialinjectionanalysissystemusingpnitrophenylbutyrate
AT rodriguezjorge monitoringlipaseesteraseactivitybystoppedflowinasequentialinjectionanalysissystemusingpnitrophenylbutyrate
AT valerofrancisco monitoringlipaseesteraseactivitybystoppedflowinasequentialinjectionanalysissystemusingpnitrophenylbutyrate
AT baezamireia monitoringlipaseesteraseactivitybystoppedflowinasequentialinjectionanalysissystemusingpnitrophenylbutyrate
AT fematricardo monitoringlipaseesteraseactivitybystoppedflowinasequentialinjectionanalysissystemusingpnitrophenylbutyrate
AT camachorosa monitoringlipaseesteraseactivitybystoppedflowinasequentialinjectionanalysissystemusingpnitrophenylbutyrate
AT sandovalgeorgina monitoringlipaseesteraseactivitybystoppedflowinasequentialinjectionanalysissystemusingpnitrophenylbutyrate
AT herreralopezenriquej monitoringlipaseesteraseactivitybystoppedflowinasequentialinjectionanalysissystemusingpnitrophenylbutyrate