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 NH3 emis...
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ftdoajarticles:oai:doaj.org/article:d0c4cb0067c14b7e8247ff8aeebf8c9d 2023-05-15T15:52:48+02:00 Development of a M9‐based urea medium (M9U) for sensitive and real‐time monitoring of ureolytic activity of bacteria and cell‐free urease Jens Jakob Sigurdarson Simon Svane Henrik Karring 2020-03-01T00:00:00Z https://doi.org/10.1002/mbo3.976 https://doaj.org/article/d0c4cb0067c14b7e8247ff8aeebf8c9d EN eng Wiley https://doi.org/10.1002/mbo3.976 https://doaj.org/toc/2045-8827 2045-8827 doi:10.1002/mbo3.976 https://doaj.org/article/d0c4cb0067c14b7e8247ff8aeebf8c9d MicrobiologyOpen, Vol 9, Iss 3, Pp n/a-n/a (2020) ammonia high throughput inhibitor plate reader assay urease Microbiology QR1-502 article 2020 ftdoajarticles https://doi.org/10.1002/mbo3.976 2022-12-31T14:26:55Z 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 NH3 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 Directory of Open Access Journals: DOAJ Articles MicrobiologyOpen 9 3 |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
language |
English |
topic |
ammonia high throughput inhibitor plate reader assay urease Microbiology QR1-502 |
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ammonia high throughput inhibitor plate reader assay urease Microbiology QR1-502 Jens Jakob Sigurdarson Simon Svane Henrik Karring Development of a M9‐based urea medium (M9U) for sensitive and real‐time monitoring of ureolytic activity of bacteria and cell‐free urease |
topic_facet |
ammonia high throughput inhibitor plate reader assay urease Microbiology QR1-502 |
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 NH3 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. |
format |
Article in Journal/Newspaper |
author |
Jens Jakob Sigurdarson Simon Svane Henrik Karring |
author_facet |
Jens Jakob Sigurdarson Simon Svane Henrik Karring |
author_sort |
Jens Jakob Sigurdarson |
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 |
https://doi.org/10.1002/mbo3.976 https://doaj.org/article/d0c4cb0067c14b7e8247ff8aeebf8c9d |
genre |
Carbonic acid |
genre_facet |
Carbonic acid |
op_source |
MicrobiologyOpen, Vol 9, Iss 3, Pp n/a-n/a (2020) |
op_relation |
https://doi.org/10.1002/mbo3.976 https://doaj.org/toc/2045-8827 2045-8827 doi:10.1002/mbo3.976 https://doaj.org/article/d0c4cb0067c14b7e8247ff8aeebf8c9d |
op_doi |
https://doi.org/10.1002/mbo3.976 |
container_title |
MicrobiologyOpen |
container_volume |
9 |
container_issue |
3 |
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1766387901318299648 |