Bioluminescent System of Luminous Bacteria for Detection of Microbial Contamination
Uloženo v:
| Název: | Bioluminescent System of Luminous Bacteria for Detection of Microbial Contamination |
|---|---|
| Autoři: | Maria A. Kirillova, Elena N. Esimbekova, Rajeev, Ranjan, Irina G. Torgashina, Valentina A. Kratasyuk |
| Přispěvatelé: | Институт фундаментальной биологии и биотехнологии, Кафедра биофизики |
| Zdroj: | Журнал Сибирского федерального университета: Серия Биология, Vol 11, Iss 2, Pp 174-180 (2018) |
| Informace o vydavateli: | Siberian Federal University, 2018. |
| Rok vydání: | 2018 |
| Témata: | bacterial luciferase, QH301-705.5, бактериальная люцифераза, 02 engineering and technology, 01 natural sciences, 6. Clean water, 0104 chemical sciences, 3. Good health, bacterial contamination, NADH:FMN-oxidoreductase, Escherichia coli, NADH:FMN-оксидоредуктаза, бактериальное загрязнение, Biology (General), 0210 nano-technology, TP248.13-248.65, 34.17, Biotechnology |
| Popis: | Microbial contamination is usually analyzed using luciferin-luciferase system of fireflies by the detection of adenosine-5’-triphosphate (ATP). There is an opportunity to assess the bacterial contamination of various objects based on a quantitative analysis of other nucleotides. In the present study, a bioluminescent enzyme system of luminous bacteria NADH:FMN-oxidoreductase (Red) and luciferase (BLuc) was investigated to understand if it can be used for quantitative measurements of bacterial cells by nicotinamide adenine dinucleotide (NADH) and flavin mononucleotide (FMN) detection. To increase the sensitivity of bioluminescent system to FMN and NADH, optimization of assay conditions was performed by varying enzymes and substrates concentrations. The lowest limits of detection were 1.2 nM FMN and 0.1 pM NADH. Escherichia coli cells were used as a model bacterial sample. FMN and NADH extraction was made by destructing cell membrane by ultrasonication. Cell suspension was added into the reaction mixture instead of FMN and NADH, and light intensity depended on number of bacterial cells in the reaction mixture. Centrifugation of sonicated sample as an additional step of sample preparation did not improve the sensitivity of method. The experimental results showed that Red and BLuc system could detect at least 800 thousand bacterial cells mL-1 by determining concentration of NADH extracted from lysed cells, while 3.9 million cells mL-1 can be detected by determining concentration of FMN |
| Druh dokumentu: | Article |
| ISSN: | 2313-5530 1997-1389 |
| DOI: | 10.17516/1997-1389-0060 |
| Přístupová URL adresa: | http://elib.sfu-kras.ru/bitstream/2311/71720/1/09_Kirillova.pdf https://doaj.org/article/1bd2c35643e5496d854f512fcf825375 https://openrepository.ru/article?id=455695 http://elib.sfu-kras.ru/bitstream/2311/71720/1/09_Kirillova.pdf http://elib.sfu-kras.ru/handle/2311/71720 |
| Rights: | CC BY NC |
| Přístupové číslo: | edsair.doi.dedup.....3120e66b2fc3c59e2461962d86ce996c |
| Databáze: | OpenAIRE |
Full Text Finder 
Nájsť tento článok vo Web of Science | Abstrakt: | Microbial contamination is usually analyzed using luciferin-luciferase system of fireflies by the detection of adenosine-5’-triphosphate (ATP). There is an opportunity to assess the bacterial contamination of various objects based on a quantitative analysis of other nucleotides. In the present study, a bioluminescent enzyme system of luminous bacteria NADH:FMN-oxidoreductase (Red) and luciferase (BLuc) was investigated to understand if it can be used for quantitative measurements of bacterial cells by nicotinamide adenine dinucleotide (NADH) and flavin mononucleotide (FMN) detection. To increase the sensitivity of bioluminescent system to FMN and NADH, optimization of assay conditions was performed by varying enzymes and substrates concentrations. The lowest limits of detection were 1.2 nM FMN and 0.1 pM NADH. Escherichia coli cells were used as a model bacterial sample. FMN and NADH extraction was made by destructing cell membrane by ultrasonication. Cell suspension was added into the reaction mixture instead of FMN and NADH, and light intensity depended on number of bacterial cells in the reaction mixture. Centrifugation of sonicated sample as an additional step of sample preparation did not improve the sensitivity of method. The experimental results showed that Red and BLuc system could detect at least 800 thousand bacterial cells mL-1 by determining concentration of NADH extracted from lysed cells, while 3.9 million cells mL-1 can be detected by determining concentration of FMN |
|---|---|
| ISSN: | 23135530 19971389 |
| DOI: | 10.17516/1997-1389-0060 |