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*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

49 Downloads (Pure)

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.

Original languageEnglish
Article numbere976
JournalMicrobiologyOpen
Volume9
Issue number3
Number of pages11
DOIs
Publication statusPublished - Mar 2020

Keywords

  • ammonia
  • high throughput
  • inhibitor
  • plate reader assay
  • urease

Fingerprint Dive into the research topics of 'Development of a M9-based urea medium (M9U) for sensitive and real-time monitoring of ureolytic activity of bacteria and cell-free urease'. Together they form a unique fingerprint.

Cite this