Comparison of the spectral emission of lux recombinant and bioluminescent marine bacteria

Abstract

The purpose of the present paper was to study the influence of bacteria harbouring the luciferase-encoding Vibrio harveyi luxAB genes upon the spectral emission during growth in batch-culture conditions. In vivo bioluminescence spectra were compared from several bioluminescent strains, either naturally luminescent (Vibrio fischeri and Vibrio harveyi) or in recombinant strains (two Gram-negative Escherichia coli::luxAB strains and a Gram-positive Bacillus subtilis::luxAB strain). Spectral emission was recorded from 400 nm to 750 nm using a highly sensitive spectrometer initially devoted to Raman scattering. Two peaks were clearly identified, one at 491-500 nm (+/- 5 nm) and a second peak at 585-595 (+/- 5 nm) with the Raman CCD. The former peak was the only one detected with traditional spectrometers with a photomultiplier detector commonly used for spectral emission measurement, due to their lack of sensitivity and low resolution in the 550-650 nm window. When spectra were compared between all the studied bacteria, no difference was observed between natural or recombinant cells, between Gram-positive and Gram-negative strains, and growth conditions and growth medium were not found to modify the spectrum of light emission.

Simultaneous Evaluation of Chemiluminescence and Bioluminescence in a Phagocytic System

“…This indicated the presence of emission in a longer-wave (up to 600 nm) band. This spectrum is typical of the majority of natural and recombinant fluorescent microorganisms [15], utilizing 4a-hydroxyflavin as the photoemitter; its degradation is associated with release of excessive energy (blue-green light quantum) [12].…”

Reporter Proteins in Whole-Cell Optical Bioreporter Detection Systems, Biosensor Integrations, and Biosensing Applications

“…There is a secondary emission peak at 590 nm, however, this is only detectable using highly sensitive Raman scattering [44]. The natural aldehyde for this reaction is believed to be tetradecanal, however, the enzyme is capable of functioning with alternative aldehydes as substrates [36].…”

New bioreactor for in situ simultaneous measurement of bioluminescence and cell density

“…21 The more toxic the compounds tested, the greater the decrease in bioluminescence. During growth inside the bioreactor, light emitted from the bioluminescent reaction, which corresponds to a broad emission band in the visible range, 22 is collected with the lens assembly and focused into the glass fiber bundle with a numerical aperture compatible with that of the fiber bundle. The entire aperture of the bundle does not collect light since the fiber bundle is made up of small cylindrical fibers packed together.…”

“…At 670 nm, we have T opt ϭ0. 22. The noise power due to the electronic setup was calculated as V ES ϭ5.1ϫ10 Ϫ8 V/Hz 1/2 , with bandwidth ⌬ f ϭ f e /2.…”

“…On the other hand, indirect methods use several reaca͒ Author to whom correspondence should be addressed; electronic mail: pascal.picart@univ-lemans.fr tions such as luminescence, impedance, potentiometry, or the measurement of cell components such as ATP, proteins, lipids, and nucleic acids. [3][4][5][6][7] Rapid reliable quantitative methods for on-line measurement of biomass have been developed over the last decade. Microcalorimetric methods are based on heat evolution during growth and this correlates well to the concentration of the biomass.…”