Application of Enzyme Bioluminescence in Ecology

Abstract

: 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.

Use of Bioluminescent Saliva Testing in Evaluating of Physical Preparedness of Athletes With Different Qualifications

“…Эффективное экспрессное лабораторное диагно-стирование влияния физической нагрузки на организм спортсмена можно проводить по слюне с использовани-ем биолюминесцентного метода. Биолюминесцентный метод, основанный на изменении (уменьшении или уве-личении) интенсивности свечения биолюминесцентной ферментативной реакции в ответ на добавление анали-зируемых образцов, разработан для оценки качества, загрязнения образцов и апробирован нами ранее для выявления степени эндотоксикоза у пациентов при раз-личных заболеваниях [12,13,14,15]. Предварительные исследования показали, что биолюминесцентый метод может быть использован также для мониторинга биохи-мических изменений в слюне спортсменов, как во время физических нагрузок, так и в восстановительный период, и позволяет оптимизировать тренировочный процесс, предупреждать перегрузки для достижения спортсмена-ми рекордных результатов [16].…”

On the mechanism of biological activation by tritium

“…These are the reasons why luminous bacteria have been used as a toxicity bioassay for several decades (Grabert and Kossler, 1997;Roda et al, 2004;Girotti et al, 2008;Thakur and Ragavan, 2013;Esimbekova et al, 2014;Kudryasheva and Tarasova, 2015). It also explains the increasing interest and intensive research into the functions of luminous bacteria, their sensitivity to exogenous compounds (Girotti et al 2008;Deryabin and Aleshina, 2008;Hastings, 2012), and the mechanisms of light emission (Nemtseva and Kudryasheva, 2007;Hou et al, 2014).…”

Contrasting relationship between macro- and microviscosity of the gelatin- and starch-based suspensions and gels

“…Currently biological polymers gelatin and starch are widely used in biotechnology in media for enzyme immobilization [1][2][3][4][5] or in responsive polymer systems for controlled drug delivery [6]. Gelatin and starch gels meet many requirements for media suitable for immobilization of the biological components (high mechanical strength, porosity, etc.).…”

“…Important physico-chemical characteristic of gelatin and starch solutions and gels is the increased viscosity, which affects the rate of diffusion of both the immobilized (entrapped) components and analyte molecules. Viscosity change can influence the functional activity of proteins included into the gel, as well as the rate of drug release [2][3][4][5][6].…”

“…The aim of the current study was to analyze the relationship between macro-and microviscosity of solutions and gels based on potato starch and gelatin that were used for immobilization of enzymes from luminous bacteria [2][3][4][5]. The increasing enzyme activity and stability were revealed at certain concentrations of biopolymers [2][3][4][5].…”