Bioluminescent Bioassays Based on Luminous Bacteria

K., Rodicheva Emma; M., Kuznetsov Alexander; A., Tyulkova Nataliya; E., Medvedeva Svetlana

doi:10.17516/1997-1389-0222

Cited by 14 publications

References 127 publications

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The use of bioluminescent enzyme bioassay for the analysis of human saliva: Advantages and disadvantages

Kolenchukova,

Dedora,

Stepanova

et al. 2024

Luminescence

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The purpose of the work was to find optimal conditions for bioluminescent enzymatic analysis of saliva (based on the use of NADH:FMN oxidoreductase + luciferase) and then to determine the biological effect of using bioluminescence assay of saliva to study the physiological state of the body under normal and pathological conditions. The saliva of snowboarders and students were studied in the “rest–training” model. The saliva of patients diagnosed with acute pharyngitis was examined in the “sick–healthy” model. Bioluminescence assay was performed with a lyophilized and immobilized bi‐enzyme system using cuvette, plate, and portable luminometers. The concentrations of secretory immunoglobulin A (sIgA) and cortisol were determined by enzyme immunoassay, and the total protein content was measured by spectrophotometric method. The activity of the bioluminescent system enzymes increased as the amount and volume of saliva in the sample was decreased. The cuvette and plate luminometers were sensitive to changes in the luminescence intensity in saliva assay. Luminescence intensity correlated with the concentrations of sIgA and cortisol. The integrated bioluminescent index for saliva was reduced in the “rest–training” model and increased in the “sick–healthy” model. Thus, the non‐invasive bioluminescent saliva analysis may be a promising tool for assessing the health of the population.

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The use of bioluminescent enzyme bioassay for the analysis of human saliva: Advantages and disadvantages

Kolenchukova,

Dedora,

Stepanova

et al. 2024

Luminescence

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Application of Enzyme Bioluminescence in Ecology

Esimbekova

Kratasyuk

Shimomura

2014

Advances in Biochemical Engineering/Biotechnology

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: This review examines the general principles of bioluminescent enzymatic toxicity bioassays and describes the applications of these methods and the implementation in commercial biosensors. Bioluminescent enzyme system technology (BEST) has been proposed in the bacterial coupled enzyme system, wherein NADH:FMN-oxidoreductase-luciferase substitutes for living organisms. BEST was introduced to facilitate and accelerate the development of cost-competitive enzymatic systems for use in biosensors for medical, environmental, and industrial applications. For widespread use of BEST, the multicomponent reagent "Enzymolum" has been developed, which contains the bacterial luciferase, NADH:FMN-oxidoreductase, and their substrates, co-immobilized in starch or gelatin gel. Enzymolum is the central part of Portable Laboratory for Toxicity Detection (PLTD), which consists of a biodetector module, a sampling module, a sample preparation module, and a reagent module. PLTD instantly signals chemical-biological hazards and allows us to detect a wide range of toxic substances. Enzymolum can be integrated as a biological module into the portable biodetector-biosensor originally constructed for personal use. Based on the example of Enzymolum and the algorithm for creating new enzyme biotests with tailored characteristics, a new approach was demonstrated in biotechnological design and construction. The examples of biotechnological design of various bioluminescent methods for ecological monitoring were provided. Possible applications of enzyme bioassays are seen in the examples for medical diagnostics, assessment of the effect of physical load on sportsmen, analysis of food additives, and in practical courses for higher educational institutions and schools. The advantages of enzymatic assays are their rapidity (the period of time required does not exceed 3-5 min), high sensitivity, simplicity and safety of procedure, and possibility of automation of ecological monitoring; the required luminometer is easily available.

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