Development of a flow injection analytical system for short chain amide determination based on a tubular bioreactor and an ammonium sensor

The Analyst
Marta I S VeríssimoM Teresa S R Gomes

Abstract

Pseudomonas aeruginosa (P. aeruginosa) possesses intracellular amidase activity, which catalyses the hydrolysis of short aliphatic amides producing NH4+, and has already been used along with an ammonium ion selective electrode for amide quantification. However, the incorporation of a biological membrane turned to be a challenging process and either the final arrangement was prone to amidase losses or the recovery of the sensor coating after the interaction took too long. In this article a flow injection system with an ammonium acoustic wave sensor is proposed, and after testing several different arrangements for the biological element, the ultimate choice consisted of the immobilization of a P. aeruginosa cell-free extract in the inner wall of a tubular glass reactor, which resulted in a reliable analytical system. Response times less than one minute and complete recovery in less than two minutes assured conveniently fast analysis. The analytical system, as long as the column was properly stored in HEPES buffer containing 2 mM β-mercaptoethanol and 1 mM benzamidine and refrigerated when not in use, could be used at least for 20 working days, along a period of one month, maintaining the initial sensitivity.

References

Sep 28, 2006·Analytical and Bioanalytical Chemistry·Nagendra Kumar KamisettyKeisuke Makino
Apr 20, 2007·The Journal of Biological Chemistry·Jorge AndradeCarlos Frazão
Dec 1, 2012·Enzyme Research·Ameel M R Al-Mayah
Mar 28, 2013·Chemical Society Reviews·Roger A Sheldon, Sander van Pelt
Apr 24, 2013·Enzyme and Microbial Technology·Nesrine AissaouiJean-François Lambert

❮ Previous
Next ❯

Methods Mentioned

BETA
biosensor

Software Mentioned

LabView

Related Concepts

Related Feeds

Bioreactors (ASM)

Bioreactors are important devices or systems that utilize living cells or enzymes for a chemical process. These devices carefully control and monitor the environment factors such as pH, nutrients level, and temperature, which can affect the yield in a given process. Discover the latest research here.

© 2022 Meta ULC. All rights reserved