Influence of the luxR Regulatory Gene Dosage and Expression Level on the Sensitivity of the Whole-Cell Biosensor to Acyl-Homoserine Lactone
| Autor: | Uliana S Novoyatlova, Vadim Fomin, Vladimir A. Chistyakov, O. E. Melkina, Svetlana Khrulnova, Ilya V. Manukhov, Ekaterina Scheglova, Sergey V. Bazhenov |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Aliivibrio
Clinical Biochemistry Homoserine lac operon Biosensing Techniques Acyl-Butyrolactones autoinducer Article 03 medical and health sciences chemistry.chemical_compound 4-Butyrolactone Genes Regulator Escherichia coli Aliivibrio fischeri Promoter Regions Genetic Gene 030304 developmental biology Regulator gene 0303 health sciences biology 030306 microbiology Chemistry food and beverages whole-cell biosensor quorum sensing General Medicine biochemical phenomena metabolism and nutrition biology.organism_classification Quorum sensing Biochemistry Trans-Activators bacteria Autoinducer LuxR TP248.13-248.65 Biotechnology |
| Zdroj: | Biosensors, Vol 11, Iss 166, p 166 (2021) Biosensors Volume 11 Issue 6 |
| ISSN: | 2079-6374 |
| Popis: | Aliivibrio fischeri LuxR and Aliivibrio logei LuxR1 and LuxR2 regulatory proteins are quorum sensing transcriptional (QS) activators, inducing promoters of luxICDABEG genes in the presence of an autoinducer (3-oxo-hexanoyl-l-homoserine lactone). In the Aliivibrio cells, luxR genes are regulated by HNS, CRP, LitR, etc. Here we investigated the role of the luxR expression level in LuxI/R QS system functionality and improved the whole-cell biosensor for autoinducer detection. Escherichia coli-based bacterial lux-biosensors were used, in which Photorhabdus luminescensluxCDABE genes were controlled by LuxR-dependent promoters and luxR, luxR1, or luxR2 regulatory genes. We varied either the dosage of the regulatory gene in the cells using additional plasmids, or the level of the regulatory gene expression using the lactose operon promoter. It was shown that an increase in expression level, as well as dosage of the regulatory gene in biosensor cells, leads to an increase in sensitivity (the threshold concentration of AI is reduced by one order of magnitude) and to a two to threefold reduction in response time. The best parameters were obtained for a biosensor with an increased dosage of luxRA. fischeri (sensitivity to 3-oxo-hexanoyl-l-homoserine lactone reached 30–100 pM). |
| Databáze: | OpenAIRE |
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