Development of a M9‐based urea medium (M9U) for sensitive and real‐time monitoring of ureolytic activity of bacteria and cell‐free urease
Abstract The enzyme urease is widespread in nature and catalyzes the hydrolysis of urea to form ammonia and carbonic acid. The high proficiency of the enzyme is associated with a wide range of societal challenges. In agriculture, bacterial urease activity leads to loss of fertilizer through NH 3 emi...
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crwiley:10.1002/mbo3.976 2024-06-23T07:52:03+00:00 Development of a M9‐based urea medium (M9U) for sensitive and real‐time monitoring of ureolytic activity of bacteria and cell‐free urease Sigurdarson, Jens Jakob Svane, Simon Karring, Henrik The Green Development and Demonstration Programme, GUDP, under The Danish Ministry of Environment and Food. 2020 http://dx.doi.org/10.1002/mbo3.976 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fmbo3.976 https://onlinelibrary.wiley.com/doi/pdf/10.1002/mbo3.976 https://onlinelibrary.wiley.com/doi/full-xml/10.1002/mbo3.976 en eng Wiley http://creativecommons.org/licenses/by/4.0/ MicrobiologyOpen volume 9, issue 3 ISSN 2045-8827 2045-8827 journal-article 2020 crwiley https://doi.org/10.1002/mbo3.976 2024-06-13T04:19:41Z Abstract The enzyme urease is widespread in nature and catalyzes the hydrolysis of urea to form ammonia and carbonic acid. The high proficiency of the enzyme is associated with a wide range of societal challenges. In agriculture, bacterial urease activity leads to loss of fertilizer through NH 3 emission, which has a negative impact on the environment and human health. Urease is also an essential virulence factor for several pathogenic bacteria. To screen for potential urease inhibitors, efficient, sensitive, and accurate urease activity assays are needed. However, most urease activity assays are labor‐intensive and become time‐consuming when used to screen multiple samples. Based on systematic optimization, we have developed a urea‐containing growth medium and method for continuous real‐time monitoring and screening of urease activity from both bacterial cells and pure urease in a plate reader setup. The defined M9‐based urea (M9U) medium was found to be more sensitive and suitable for a plate reader setup than both Christensen's urea broth (CUB) and Stuart's urea broth (SUB), which are established and well‐known complex urea media that formed the principle foundation of M9U. Furthermore, we show that urease activity measurements using the M9U medium in our plate reader‐based method allow reliable high‐throughput screening of urease inhibitors. Article in Journal/Newspaper Carbonic acid Wiley Online Library MicrobiologyOpen 9 3 |
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English |
description |
Abstract The enzyme urease is widespread in nature and catalyzes the hydrolysis of urea to form ammonia and carbonic acid. The high proficiency of the enzyme is associated with a wide range of societal challenges. In agriculture, bacterial urease activity leads to loss of fertilizer through NH 3 emission, which has a negative impact on the environment and human health. Urease is also an essential virulence factor for several pathogenic bacteria. To screen for potential urease inhibitors, efficient, sensitive, and accurate urease activity assays are needed. However, most urease activity assays are labor‐intensive and become time‐consuming when used to screen multiple samples. Based on systematic optimization, we have developed a urea‐containing growth medium and method for continuous real‐time monitoring and screening of urease activity from both bacterial cells and pure urease in a plate reader setup. The defined M9‐based urea (M9U) medium was found to be more sensitive and suitable for a plate reader setup than both Christensen's urea broth (CUB) and Stuart's urea broth (SUB), which are established and well‐known complex urea media that formed the principle foundation of M9U. Furthermore, we show that urease activity measurements using the M9U medium in our plate reader‐based method allow reliable high‐throughput screening of urease inhibitors. |
author2 |
The Green Development and Demonstration Programme, GUDP, under The Danish Ministry of Environment and Food. |
format |
Article in Journal/Newspaper |
author |
Sigurdarson, Jens Jakob Svane, Simon Karring, Henrik |
spellingShingle |
Sigurdarson, Jens Jakob Svane, Simon Karring, Henrik Development of a M9‐based urea medium (M9U) for sensitive and real‐time monitoring of ureolytic activity of bacteria and cell‐free urease |
author_facet |
Sigurdarson, Jens Jakob Svane, Simon Karring, Henrik |
author_sort |
Sigurdarson, Jens Jakob |
title |
Development of a M9‐based urea medium (M9U) for sensitive and real‐time monitoring of ureolytic activity of bacteria and cell‐free urease |
title_short |
Development of a M9‐based urea medium (M9U) for sensitive and real‐time monitoring of ureolytic activity of bacteria and cell‐free urease |
title_full |
Development of a M9‐based urea medium (M9U) for sensitive and real‐time monitoring of ureolytic activity of bacteria and cell‐free urease |
title_fullStr |
Development of a M9‐based urea medium (M9U) for sensitive and real‐time monitoring of ureolytic activity of bacteria and cell‐free urease |
title_full_unstemmed |
Development of a M9‐based urea medium (M9U) for sensitive and real‐time monitoring of ureolytic activity of bacteria and cell‐free urease |
title_sort |
development of a m9‐based urea medium (m9u) for sensitive and real‐time monitoring of ureolytic activity of bacteria and cell‐free urease |
publisher |
Wiley |
publishDate |
2020 |
url |
http://dx.doi.org/10.1002/mbo3.976 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fmbo3.976 https://onlinelibrary.wiley.com/doi/pdf/10.1002/mbo3.976 https://onlinelibrary.wiley.com/doi/full-xml/10.1002/mbo3.976 |
genre |
Carbonic acid |
genre_facet |
Carbonic acid |
op_source |
MicrobiologyOpen volume 9, issue 3 ISSN 2045-8827 2045-8827 |
op_rights |
http://creativecommons.org/licenses/by/4.0/ |
op_doi |
https://doi.org/10.1002/mbo3.976 |
container_title |
MicrobiologyOpen |
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9 |
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3 |
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1802643258234372096 |