Correlation between cellular fluorescence and NAD(P)H levels in Alcaligenes eutrophus JMP 134

Simon, D.; Müller, Roland; Bley, Thomas; Babel, Wolfgang

doi:10.1007/s004490050183

Cited by 2 publications

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“…Culture broths are known to contain extracellular¯uoro-phores [16,18,19,28,29]. Figure 11 shows the time pro®le of NAD(P)H-like¯uorescence and¯avin-like¯uorescence measured on-line in the cell-free medium during nutristat cultivation at a phenol concentration of 550±650 mg/l.…”

Section: Fluorescence Of Cell-free Mediummentioning

confidence: 99%

Response of Ralstonia eutropha JMP 134 to long-term exposure to toxic substrates in nutristat cultivation as indicated by on-line fluorescence measurements

Simon

Müller

Grosse

et al. 2000

Bioprocess Engineering

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Ralstonia eutropha JMP 134 was continuously grown on phenol and 2,4-dichlorophenoxyacetate under nutristatic conditions at elevated stationary concentrations of 90±650 mg phenol/l and 25±100 mg 2,4-D/l, respectively, in order to study the response of the bacterial population to long-term exposure to these potentially toxic substrates. The course of the cells' response over time was observed by determining distinctive growth parameters and by the on-line measurement of¯uorescence spectra of intracellular and extracellular¯uorophores. The latter were monitored using a modi®ed¯uorescence spectrophotometer. The results of the nutristat experiments indicate that the adaptation of the culture to long-term exposure to phenol and 2,4-D exhibited dynamic characteristics of the growth pattern determined by the individual substrates and their concentration, including enforced and reduced levels of substrate conversion. This growth pattern is interpreted as an expression of superimposing cellular events in order to withstand unfavorable environmental conditions. Finally, the growth rate attained retarded levels under stationary conditions, slowing down to almost zero for example in the case of about 100 mg 2,4-D/l.The growth rate pro®le within the various phases of adaptation was well re¯ected by the¯uorescence signals. The NAD(P)H¯uorescence was almost exclusively emitted by the cellular pool of NADPH and behaved inversely to the growth rate. A similar relationship was obtained for the cellular¯uorescence of a¯avin-containing compound. Sharply reduced growth was additionally accompanied by a rapid rise of the background¯uorescence. These data indicate that¯uorescence-derived signals provide a useful re¯ection of cellular events in inhibited growth situations.

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Section: Fluorescence Of Cell-free Mediummentioning

confidence: 99%

Response of Ralstonia eutropha JMP 134 to long-term exposure to toxic substrates in nutristat cultivation as indicated by on-line fluorescence measurements

Simon

Müller

Grosse

et al. 2000

Bioprocess Engineering

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“…Duysens and Amesz first reported the use of on-line fluorescence measurement technique in suspensions of baker's yeast and alga [15]. Thereafter it has been used by several researchers for in situ characterization of biomass and cellular metabolic state using different fermentation processes [13,[16][17][18][19]. The correlation between culture fluorescence and biomass concentration could be made on a reasonable assumption that during the balanced growth of the microorganism the intracellular concentration of NADH+H + remains constant.…”

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confidence: 98%

On-line Characterization of Metabolic State in Batch Cultivation of Clostridium diolis for 1,3-Propanediol Production Using NADH+H+ Fluorescence

Kaur

Sharma

Srivastava

et al. 2011

Appl Biochem Biotechnol

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NADH is a coenzyme which plays a central role in cellular growth and metabolism. It is an intracellular fluorophore which fluoresces at 460 nm when cells are irradiated by 340 nm wavelength of light. The application of NADH+H(+) fluorescence measurement for characterization of biomass and its metabolic activity during batch fermentation of 1,3-propanediol (1,3-PD) using Clostridium diolis was investigated in this study. A linear correlation between net fluorescence and biomass concentration was observed during both the initial and final phases of 1,3-PD fermentation. This could be used as an on-line indicator of biomass concentration inside the bioreactor thereby eliminating the need for sampling and off-line analysis for establishing biomass concentration during these phases. Also a sharp decline in the NADH+H(+) fluorescence value was obtained towards the end of fermentation which could be a significant on-line, in situ signal of substrate depletion in the bioreactor and therefore possible fresh nutrient feed for enhanced production of 1,3-PD by repetitive and/or various fed-batch cultivation(s). This is the first report on the use of NADH + H(+) fluorescence measurement technique for 1,3-PD fermentation.

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Correlation between cellular fluorescence and NAD(P)H levels in Alcaligenes eutrophus JMP 134

Cited by 2 publications

References 0 publications

Response of Ralstonia eutropha JMP 134 to long-term exposure to toxic substrates in nutristat cultivation as indicated by on-line fluorescence measurements

Response of Ralstonia eutropha JMP 134 to long-term exposure to toxic substrates in nutristat cultivation as indicated by on-line fluorescence measurements

On-line Characterization of Metabolic State in Batch Cultivation of Clostridium diolis for 1,3-Propanediol Production Using NADH+H+ Fluorescence

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