RESUMO
The design and performance characteristics of a small volume (20 ml) continuous culture device for the cultivation of luminous bacteria are described. This simply constructed device can be used to supply luminescent bacteria with constant properties for either laboratory use or the assay of environmental pollutants. Furthermore, bacteria can be deployed to make sensitive (<1 nM) oxygen measurements. The culture device may be configured, alone or in combination, as a chemostat, turbidostat or a "bioluminostat" where bacterial bioluminescence becomes the controlling variable. Over extended periods (>1 week) it was possible to maintain steady-state luminescence within 5% of a pre-set value, although occasionally a non-bioluminescent "mutant" became dominant; in this case light emission was irreversibly lost. A secondary chamber provided additional flexibility and easy manipulation of the cultivated bacteria for testing. The continuous culture system is also suitable for the growth of recombinant microorganisms that either constitutively express luciferase, or do so in response to stress promoter activity. The non-standard control methodologies reported may have important research and industrial applications, for example in providing immediate process control or as an inferential method to optimize biomass: product yield ratios.
Assuntos
Técnicas de Laboratório Clínico/instrumentação , Photobacterium , Aliivibrio fischeri , Técnicas Biossensoriais , Técnicas de Cultura de Células/instrumentação , Técnicas de Cultura de Células/métodos , Técnicas de Laboratório Clínico/métodos , Oxigênio/metabolismo , Photobacterium/metabolismo , Fatores de TempoRESUMO
The biological functions of light emission in bacterial bioluminescence are not always obvious, especially if the bacteria are in a free-living mode. Experimental evidence suggests that light emission confers benefit to the bacteria themselves such as through photoreactivation and involves as much as 20% of cell energy metabolism. A theoretical model shows if the effect is mediated solely by light then cells should be luminescent at both high and low cell densities, therefore raising doubt over the photoreactivation hypothesis and suggesting that another cofactor is involved. It has been postulated that bioelectromagnetics may be involved in biological processes and be involved with coordinated activity in quorate cells. The cell densities associated with autoinduction coincide with a large change in coupling efficiency in the millimeter and submillimeter spectral region. In this paper it is suggested that one function of bioluminescence is as a pump, involving millimeter and submillimeter wave coupling that is of benefit to the quorum. This may be related to the observation that millimeter wave radiation exposure has been reported to induce changes in DNA conformation and possibly gene expression. Agents that change DNA conformation in bioluminescent bacteria can cause increases in light emission. This work may have implications for electromagnetic fields as quorum-quenching agents.