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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 42 (1993), S. 295-302 
    ISSN: 0006-3592
    Keywords: intracellular pH ; bioreactors ; cultivation ; yeast ; 9-aminoacridine ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Intracellular pH (pHi) was measured on-line in a bioreactor using a fluorescent pHi indicator, 9-aminoacridine, and controlled fed-batch cultivations of yeast cells based on pHi (FB-pHi) were performed. In FB-pHi cultivations, automated glucose additions were made to the culture in response to culture pHi. The average ethanol (an-aerobic product) yield was significantly lower [0.12 g g-1 glucose in fed-batch pHi cultivations with 100 ppm glucose additions (FB-pHi-100 cultivation) vs. 0.48 g g-1 glucose in batch] and cell yield was higher (0.54 g g-1 glucose in FB-pHi-100 cultivation vs. 0.3 g g-1 glucose in batch) compared to batch cultivation. An expression has been derived to calculate changes in pHi from measured fluorescence values when the cell concentration increases during growth. Cultivations based on pHi, performed with different magnitudes of glucose addition (100, 50, and 10 ppm additions), showed that lower magnitudes of glucose addition resulted in lower ethanol yields while cell yield remained unaffected. The ratio of specific oxygen uptake rate to specific glucose uptake rate (OUR/GUR) increased with decreased in magnitude of glucose additions in FB-pHi cultivations, suggesting that the culture aerobic state was higher when the magnitude of glucose addition was lower. The average cell productivity in FB-pHi cultivations was 29% higher than in batch cultivation. Cells were also cultivated at high OUR conditions, and the results are compared with other cultivations. © 1993 John Wiley & Sons, Inc.
    Additional Material: 11 Ill.
    Type of Medium: Electronic Resource
    Library Location Call Number Volume/Issue/Year Availability
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 41 (1993), S. 118-128 
    ISSN: 0006-3592
    Keywords: intracellular pH ; 9-aminoacridine ; bioreactor ; on-line measurement ; yeast ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: A method has been developed to continuously measure the intracellular pH (pHi) 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 pHi indicator 9-aminoacridine (9A A) which is a week base. An expression has been derived to calculate changes in pHi from measured 9AA-fluorescence changes. The indicator 9AA was found t be nontoxic to yeast cells at concentrations used to measure pHi (7 μM). The fluorescence of nicotinamide adenine dinucleotide (NADH) molecules did not interfere significantly with the measurement of 9AA-fluorescence. The pHi 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 pHi, indicator (which is however unsuitable for on-line applications in a bioreactor) bis-carboxyethyl carboxy fluorescein (BCECF). The pHi of yeast under substrate starved conditions was 6.4 units. The responses of pHi of yeast cells to induced metabolic transitions were studied. Under aerobic condition, pHi 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, pHi increased by 0.1 unit following a 500-ppm glucose pulse addition. Comparison of pHi with other indicators of cellular metabolic state suggests that pHi is a cellular metabolic state indicator. © 1993 John Wiley & Sons, Inc.
    Additional Material: 14 Ill.
    Type of Medium: Electronic Resource
    Library Location Call Number Volume/Issue/Year Availability
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