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Degradation Kinetics of Clavulanic Acid in Fermentation Broths at Low Temperatures

Clavulanic acid (CA) is a β-lactam antibiotic inhibitor of β-lactamase enzymes, which confers resistance to bacteria against several antibiotics. CA is produced in submerged cultures by the filamentous Gram-positive bacterium Streptomyces clavuligerus; yield and downstream process are compromised by...

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Autores principales: Gómez-Ríos, David, Ramírez-Malule, Howard, Neubauer, Peter, Junne, Stefan, Ríos-Estepa, Rigoberto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6466556/
https://www.ncbi.nlm.nih.gov/pubmed/30658482
http://dx.doi.org/10.3390/antibiotics8010006
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author Gómez-Ríos, David
Ramírez-Malule, Howard
Neubauer, Peter
Junne, Stefan
Ríos-Estepa, Rigoberto
author_facet Gómez-Ríos, David
Ramírez-Malule, Howard
Neubauer, Peter
Junne, Stefan
Ríos-Estepa, Rigoberto
author_sort Gómez-Ríos, David
collection PubMed
description Clavulanic acid (CA) is a β-lactam antibiotic inhibitor of β-lactamase enzymes, which confers resistance to bacteria against several antibiotics. CA is produced in submerged cultures by the filamentous Gram-positive bacterium Streptomyces clavuligerus; yield and downstream process are compromised by a degradation phenomenon, which is not yet completely elucidated. In this contribution, a study of degradation kinetics of CA at low temperatures (−80, −20, 4, and 25 °C) and pH 6.8 in chemically-defined fermentation broths is presented. Samples of CA in the fermentation broths showed a fast decline of concentration during the first 5 h followed by a slower, but stable, reaction rate in the subsequent hours. A reversible-irreversible kinetic model was applied to explain the degradation rate of CA, its dependence on temperature and concentration. Kinetic parameters for the equilibrium and irreversible reactions were calculated and the proposed kinetic model was validated with experimental data of CA degradation ranging 16.3 mg/L to 127.0 mg/L. Degradation of the chromophore CA-imidazole, which is commonly used for quantifications by High Performance Liquid Chromatography, was also studied at 4 °C and 25 °C, showing a rapid rate of degradation according to irreversible first-order kinetics. A hydrolysis reaction mechanism is proposed as the cause of CA-imidazole loss in aqueous solutions.
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spelling pubmed-64665562019-04-18 Degradation Kinetics of Clavulanic Acid in Fermentation Broths at Low Temperatures Gómez-Ríos, David Ramírez-Malule, Howard Neubauer, Peter Junne, Stefan Ríos-Estepa, Rigoberto Antibiotics (Basel) Article Clavulanic acid (CA) is a β-lactam antibiotic inhibitor of β-lactamase enzymes, which confers resistance to bacteria against several antibiotics. CA is produced in submerged cultures by the filamentous Gram-positive bacterium Streptomyces clavuligerus; yield and downstream process are compromised by a degradation phenomenon, which is not yet completely elucidated. In this contribution, a study of degradation kinetics of CA at low temperatures (−80, −20, 4, and 25 °C) and pH 6.8 in chemically-defined fermentation broths is presented. Samples of CA in the fermentation broths showed a fast decline of concentration during the first 5 h followed by a slower, but stable, reaction rate in the subsequent hours. A reversible-irreversible kinetic model was applied to explain the degradation rate of CA, its dependence on temperature and concentration. Kinetic parameters for the equilibrium and irreversible reactions were calculated and the proposed kinetic model was validated with experimental data of CA degradation ranging 16.3 mg/L to 127.0 mg/L. Degradation of the chromophore CA-imidazole, which is commonly used for quantifications by High Performance Liquid Chromatography, was also studied at 4 °C and 25 °C, showing a rapid rate of degradation according to irreversible first-order kinetics. A hydrolysis reaction mechanism is proposed as the cause of CA-imidazole loss in aqueous solutions. MDPI 2019-01-17 /pmc/articles/PMC6466556/ /pubmed/30658482 http://dx.doi.org/10.3390/antibiotics8010006 Text en © 2019 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 (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Gómez-Ríos, David
Ramírez-Malule, Howard
Neubauer, Peter
Junne, Stefan
Ríos-Estepa, Rigoberto
Degradation Kinetics of Clavulanic Acid in Fermentation Broths at Low Temperatures
title Degradation Kinetics of Clavulanic Acid in Fermentation Broths at Low Temperatures
title_full Degradation Kinetics of Clavulanic Acid in Fermentation Broths at Low Temperatures
title_fullStr Degradation Kinetics of Clavulanic Acid in Fermentation Broths at Low Temperatures
title_full_unstemmed Degradation Kinetics of Clavulanic Acid in Fermentation Broths at Low Temperatures
title_short Degradation Kinetics of Clavulanic Acid in Fermentation Broths at Low Temperatures
title_sort degradation kinetics of clavulanic acid in fermentation broths at low temperatures
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6466556/
https://www.ncbi.nlm.nih.gov/pubmed/30658482
http://dx.doi.org/10.3390/antibiotics8010006
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