Cell envelope fluidity modification for an effective glutamate excretion in Corynebacterium glutamicum 2262.
1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH) was used to assess the cell envelope fluidity of Corynebacterium glutamicum 2262 during a temperature-triggered glutamate producing process. Because the fluorescence lifetime of TMA-DPH was shown to be constant all over the process, fluorescence anisotropy can be considered as a good index of cell envelope fluidity. When the temperature of the fed-batch culture was increased from 33 to 39 degrees C to induce glutamate excretion, the fluorescence anisotropy values decreased from 0.212 +/- 0.002 to 0.186 +/- 0.002 (corresponding to an increase in the cell fluidity), while the specific glutamate production rate reached its maximal value. The increase in fluidity of the C. glutamicum cell envelope was not due to a physical effect related to the temperature elevation, but rather to an alteration of the composition of the cell envelope. Using a mutant devoid of corynomycolates, significant differences in fluorescence anisotropy values were obtained compared to the wild-type strain, suggesting that TMA-DPH is mainly anchored into the corynomycomembrane. Differences in fluorescence anisotropy were also observed when the bacteria were cultivated at 33, 36, 38, and 39 degrees C in batch cultures, and a linear relationship was obtained between the maximum specific glutamate production rate and the measured fluidity. When using the glutamate non-producing variant of C. glutamicum 2262, the fluorescence anisotropy remained constant at 0.207 +/- 0.003 whatever the applied temperature shift. This suggests that the fluidity of the Corynebacteria mycomembrane plays an important role in glutamate excretion during the temperature-triggered process.
Bokas
Dimitrios
D
Laboratoire des Sciences du Génie Chimique, UPR CNRS 6811, ENSAIA, Institut National Polytechnique de Lorraine, 2 Avenue de la Forêt de Haye, BP 172, F-54505, Vandoeuvre-lès-Nancy, France. Jean-Louis.Goergen@ensaia.inpl-nancy.fr
Uy
Davin
D
Grattepanche
Franck
F
Duportail
Guy
G
Guedon
Emmanuel
E
Delaunay
Stéphane
S
Goergen
Jean-Louis
JL
D002462:Cell Membrane / Q000502:physiology; D048230:Corynebacterium glutamicum / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial / Q000235:genetics; D004161:Diphenylhexatriene / Q000031:analogs & derivatives / Q000737:chemistry; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes / Q000737:chemistry; D018698:Glutamic Acid / Q000378:metabolism*; D007700:Kinetics; D008560:Membrane Fluidity / Q000502:physiology*; D016254:Mutagenesis, Insertional; D013696:Temperature
D004269:DNA, Bacterial; D005456:Fluorescent Dyes; D004161:Diphenylhexatriene; D018698:Glutamic Acid; C033628:1-(4-(trimethylamino)phenyl)-6-phenylhexa-1,3,5-triene