Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Microbead-encapsulated luminescent bioreporter screening of P. aeruginosa via its secreted quorum sensing molecules.

Version 1 : Received: 14 June 2024 / Approved: 17 June 2024 / Online: 18 June 2024 (11:54:36 CEST)

A peer-reviewed article of this Preprint also exists.

Paul, A.A.; Kadosh, Y.S.; Kushmaro, A.; Marks, R.S. Microbead-Encapsulated Luminescent Bioreporter Screening of P. aeruginosa via Its Secreted Quorum-Sensing Molecules. Biosensors 2024, 14, 383. Paul, A.A.; Kadosh, Y.S.; Kushmaro, A.; Marks, R.S. Microbead-Encapsulated Luminescent Bioreporter Screening of P. aeruginosa via Its Secreted Quorum-Sensing Molecules. Biosensors 2024, 14, 383.

Abstract

Pseudomonas aeruginosa is an opportunistic Gram-negative bacterium that remains a prevalent clinical and environmental challenge. Quorum sensing (QS) molecules are effective biomarkers in pinpointing the presence of P. aeruginosa. This study aimed to develop a convenient-to-use whole-cell biosensor using P. aeruginosa reporters individually encapsulated within alginate-poly-L-lysine (alginate-PLL) microbeads to specifically detect the presence of bacterial autoinducers. The PLL-reinforced microbeads were prepared using a two-step method involving ionic crosslinking and subsequent coating with thin layers of PLL. The alginate-PLL beads showed good stability in the presence of a known cation scavenger (sodium citrate), which typically limits the widespread applications of calcium alginate. In media containing synthetic autoinducers, such as N-(3-oxo dodecanoyl) homoserine lactone (3-oxo-C12-HSL) and N-butanoyl-L-homoserine lactone (C4-HSL) or cell-free supernatants of planktonic or flow cell biofilm effluent of wild P. aeruginosa (PAO1), the encapsulated bacteria enabled a dose-dependent detection of the presence of these QS molecules. The prepared bioreporter beads remained stable during prolonged storage at 4 and -80 °C and were ready for on-the-spot sensing without the need for recovery. This whole-cell biosensor demonstrates the practicality of the encapsulated bioreporter for bacterial detection based on specific QS molecules.

Keywords

Alginate; Hydrogels; quorum sensing; , C4-HSL; whole-cell biosensors; Furanone C-30; autoinducers; Pseudomonas aeruginosa,; bioencapsulation

Subject

Biology and Life Sciences, Biology and Biotechnology

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