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A possible reason for the complexity of the signals produced by bioluminescent biosensors might be self-organization of the cells. In order to verify this possibility, bioluminescence images of cultures of lux gene reporter Escherichia coli were recorded for several hours after being placed into 8-10 mm diameter cylindrical containers. It was found that luminous cells distribute near the three-phase contact line, forming irregular azimuthal waves. As we show, space-time plots of quasi-one-dimensional bioluminescence measured along the contact line can be simulated by reaction-diffusion-chemotaxis equations, in which the reaction term for the cells is a logistic (autocatalytic) growth function. It was found that the growth rate of the luminous cells (~0.02 s(-1)) is >100 times higher than the growth rate of E. coli. We provide an explanation for this result by assuming that E. coli exhibits considerable respiratory flexibility (the ability of oxygen-induced switching from one metabolic pathway to another). According to the simple two-state model presented here, the number of oxic (luminous) cells grows at the expense of anoxic (dark) cells, whereas the total number of (oxic and anoxic) cells remains unchanged. It is conjectured that the corresponding reaction-diffusion-chemotaxis model for bioluminescence pattern formation can be considered as a model for the energy-taxis and metabolic self-organization in the population of the metabolically flexible bacteria under hypoxic conditions.
A possible reason for the complexity of the signals produced by bioluminescent biosensors might be self-organization of the cells. In order to verify this possibility, bioluminescence images of cultures of lux gene reporter Escherichia coli were recorded for several hours after being placed into 8-10 mm diameter cylindrical containers. It was found that luminous cells distribute near the three-phase contact line, forming irregular azimuthal waves. As we show, space-time plots of quasi-one-dimensional bioluminescence measured along the contact line can be simulated by reaction-diffusion-chemotaxis equations, in which the reaction term for the cells is a logistic (autocatalytic) growth function. It was found that the growth rate of the luminous cells (~0.02 s(-1)) is >100 times higher than the growth rate of E. coli. We provide an explanation for this result by assuming that E. coli exhibits considerable respiratory flexibility (the ability of oxygen-induced switching from one metabolic pathway to another). According to the simple two-state model presented here, the number of oxic (luminous) cells grows at the expense of anoxic (dark) cells, whereas the total number of (oxic and anoxic) cells remains unchanged. It is conjectured that the corresponding reaction-diffusion-chemotaxis model for bioluminescence pattern formation can be considered as a model for the energy-taxis and metabolic self-organization in the population of the metabolically flexible bacteria under hypoxic conditions.
Under adjusted experimental conditions, open-to-air cultures of lux gene-engineered Ralstonia eutropha (wholecell biosensors of copper) exhibit bioconvection, which accounts for fluctuating bioluminescence. The power spectrum of bioluminescence intensity fluctuations recorded from a cylindrical sample 9 mm in diameter and approximately 10 mm in height is characterized by a dominant low-frequency oscillation (with a characteristic period of approximately 8-12 min), which is occasionally accompanied by a few weaker oscillations. The corresponding spectral peaks emerge on a high-noise background. The spectra of bioluminescence intensity fluctuations qualitatively resemble the spectra of temperature or fluid velocity fluctuations in an appropriate turbulent thermal convection system. It has been suggested that in a bioconvective system, like in thermal convection systems, the emergence of oscillation reflects the large-scale convective circulation that spans the height of the cylindrical cell. The velocity of large-scale bioconvective circulation was estimated to be 37-48 microm/s. The occasional emergence of weaker-than-dominant oscillations was explained through the coexistence and interaction of the large-scale circulation with, presumably, a gene-expression-related cyclic process (with a characteristic period of approximately 25-50 min).
'Noisy' bioluminescence of an unstirred and exposed-to-air microculture of lux -gene fused Escherichia coli was interpreted, assuming two oscillatory processes: cyclic induction of the lac promoter, resulting in oscillatory reporter lux -gene expression, and oscillatory respiration in the culture. Biological support for the theoretical argument, that two rhythms can interact in cells to produce additional rhythms, is provided. These results, along with previously obtained data on oscillatory bioluminescence, indicate the deterministic nature of observed noise-like fluctuations and promising perspectives of lux gene reporter systems in investigations of ultradian rhythm.
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