Enzymes activity in a cell is determined by many factors, among which viscosity of the microenvironment plays a significant role. Various cosolvents can imitate intracellular conditions in vitro, allowing to reduce a combination of different regulatory effects. The aim of the study was to analyze the media viscosity effects on the rate constants of the separate stages of the bacterial bioluminescent reaction. Non-steady-state reaction kinetics in glycerol and sucrose solutions was measured by stopped-flow technique and analyzed with a mathematical model developed in accordance with the sequence of reaction stages. Molecular dynamics methods were applied to reveal the effects of cosolvents on luciferase structure. We observed both in glycerol and in sucrose media that the stages of luciferase binding with flavin and aldehyde, in contrast to oxygen, are diffusion-limited. Moreover, unlike glycerol, sucrose solutions enhanced the rate of an electronically excited intermediate formation. The MD simulations showed that, in comparison with sucrose, glycerol molecules could penetrate the active-site gorge, but sucrose solutions caused a conformational change of functionally important Glu175 of luciferase. Therefore, both cosolvents induce diffusion limitation of substrates binding. However, in sucrose media, increasing enzyme catalytic constant neutralizes viscosity effects. The activating effect of sucrose can be attributed to its exclusion from the catalytic gorge of luciferase and promotion of the formation of the active site structure favorable for the catalysis.
Forecasting response of the biosphere and regional ecosystems to observed and expected climate change is the fundamental problem with obvious practical significance. Fundamental non-linearity of the climate system and biosphere makes feasible implementing multiple states and threshold processes in the biosphere-climate system (BCS) in response to gradually increasing influence factor (greenhouse gas concentrations growth). Really time series analysis of global temperature and other global and local parameters indicates the presence of abrupt transitions between stationary states. Identification of the switching mechanisms using general circulation models of the atmosphere and the ocean is associated with the obvious difficulties due to their complexity. Understanding the nature of such switches at qualitative level can be achieved by using a conceptual small-scale models. Some variants of possible mechanisms capable of generating these shifts and simultaneously supporting quasi-stationary periods between them are discussed.
In accordance with the ideas of V.I. Vernadsky, the Earth’s biosphere can exist only because of the high degree of closure of the cyclic matter transformations carried out by all living organisms by using the energy from the Sun. In the course of its evolution, the Earth’s biosphere has undergone a number of cardinal transformations, but, at least for the last 20 million years, the gas composition of the atmosphere, and primarily the concentration of carbon dioxide, has remained practically unchanged. Nevertheless, the high degree of closure of material flows in the Earth’s biosphere seems paradoxical, since closure is not an adaptive feature of an individual undergoing natural selection for traits that give an advantage here and now (the Vernadsky–Darwin paradox). The stages in the formation of the closure of the Earth’s biosphere are considered in the context of four epochs that differ in the energy available to living organisms: (1) geochemical energy; (2) solar energy; (3) energy of oxidative phosphorylation; and (4) consumption of living flesh, predation. The paper considers possible options for resolving the VD paradox using as the example models of closed ecological systems (CES) with low species diversity. The fundamental inapplicability of ecological models with rigid metabolism for the description of CES is shown. Three mechanisms for resolving the VD paradox are proposed and the conditions for their implementation are assessed: (1) a stochastic mechanism: random selection of closing organisms (decomposers) with the corresponding stoichiometric ratios; (2) changing the consumption stoichiometry by switching catabolic pathways to different types of substances (proteins, fats, carbohydrates); and (3) changing the consumption stoichiometry by choosing food, depending on the state of internal nutrient pools. The present study leads to the conclusion that the Vernadsky–Darwin paradox can be resolved in nature by combining the mechanisms that simultaneously provide both a current competitive advantage and the ability to close trophic chains with a wide variation in the composition of material flows.