Use of NAD(P)H-fluorescence for monitoring the response of starved cells of Catharanthus roseus in suspension to metabolic perturbations

Asali, E. C.; Mutharasan, Raj; Humphrey, Arthur E.

doi:10.1016/0168-1656(92)90101-e

Cited by 15 publications

(1 citation statement)

References 15 publications

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“…Online monitoring of culture fluorescence from intrinsic fluorophores such as NAD(P)H, ) or recombinant fluorescent reporters such as GFP, can provide valuable information of the culture metabolic states, allowing development of improved process control strategies to increase protein production. Asali et al [93] used NAD(P)H fluorescence to monitor the response of starved Catharanthus roseus cells to metabolic perturbations. Choi et al [94] used a fibre-optic probe for on-line sensing of NAD(P)H culture fluorescence of tobacco suspension culture and correlated the fluorescence signal to biomass concentration.…”

Section: Advances In Process Monitoringmentioning

confidence: 99%

Bioreactor Engineering For Recombinant Protein Production Using Plant Cell Suspension Culture

Plan Tissue Culture Engineering

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Section: Advances In Process Monitoringmentioning

confidence: 99%

Bioreactor Engineering For Recombinant Protein Production Using Plant Cell Suspension Culture

Plan Tissue Culture Engineering

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Bioreactors, Stirred Tank

Doran

2000

Encyclopedia of Cell Technology

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Intracellular pH based controlled cultivation of yeast cells: I. Measurement methodology

Sureshkumar

Mutharasan

1993

Biotech & Bioengineering

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A method has been developed to continuously measure the intracellular pH (pH(i)) of cells cultivated in a bioreactor in an on-line fashion over extend time periods. The methods is attractive in its simplicity and involves the use of a fluorescent pH(i) indicator 9-aminoacridine (9A A) which is a week base. An expression has been derived to calculate changes in pH(i) from measured 9AA-fluorescence changes. The indicator 9AA was found t be nontoxic to yeast cells at concentrations used to measure pH(i) (7 microM). The fluorescence of nicotinamide adenine dinucleotide (NADH) molecules did not interfere significantly with the measurement of 9AA-fluorescence. The pH(i) change in yeast cell following the addition of a proton ionophore carbonyl cyanide m-chlorophenyl hydrazone (CCCP) measured by 9AA compared favorably with that measured by the well-established pH(i), indicator (which is however unsuitable for on-line applications in a bioreactor) bis-carboxyethyl carboxy fluorescein (BCECF). The pH(i) of yeast under substrate starved conditions was 6.4 units. The responses of pH(i) of yeast cells to induced metabolic transitions were studied. Under aerobic condition, pH(i) increased by 0.12 unit following a 100-ppm glucose pulse addition and by 0.25 unit following a 300-ppm ethanol pulse addition. Under anaerobic condition, pH(i) increased by 0.1 unit following a 500-ppm glucose pulse addition. Comparison of pH(i) with other indicators of cellular metabolic state suggests that pH(i) is a cellular metabolic state indicator.

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Use of NAD(P)H-fluorescence for monitoring the response of starved cells of Catharanthus roseus in suspension to metabolic perturbations

Cited by 15 publications

References 15 publications

Bioreactor Engineering For Recombinant Protein Production Using Plant Cell Suspension Culture

Bioreactor Engineering For Recombinant Protein Production Using Plant Cell Suspension Culture

Bioreactors, Stirred Tank

Intracellular pH based controlled cultivation of yeast cells: I. Measurement methodology

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