Monitoring lipase/esterase activity by stopped flow in a sequential injection analysis system using p-nitrophenyl butyrate

Jorge Pliego, Juan Carlos Mateos, Jorge Rodriguez, Francisco Valero, Mireia Baeza, Ricardo Femat, Rosa Camacho, Georgina Sandoval, Enrique J. Herrera-López

Research output: Contribution to journalArticleResearchpeer-review

23 Citations (Scopus)


© 2015 by the authors; licensee MDPI, Basel, Switzerland. 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.
Original languageEnglish
Pages (from-to)2798-2811
Publication statusPublished - 27 Jan 2015


  • Lipase/esterase activity
  • Monitoring
  • P-nitrophenyl esters
  • Sequential injection analysis
  • Stopped flow


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