University of North Florida
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Stuart Chalk, Ph.D.
Department of Chemistry
University of North Florida
Phone: 1-904-620-1938
Fax: 1-904-620-3535
Website: @unf

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Classification: Cell -> saccharomyces cerevisiae

Citations 2

"Determination Of Intracellular Trehalose And Glycogen In Saccharomyces Cerevisiae"
Anal. Biochem. 1995 Volume 228, Issue 1 Pages 143-149
Schulze U., Larsen M. E. and Villadsen J.

Abstract: Saccharomyces cerevisiae cells were suspended in 1 mL 40 mM potassium acetate of pH 4.8. The cells were disintegrated in a bead mill at a vibration frequency of 150-1800/min for 30 min at 4°C. After centrifugation, the supernatant solution was hydrolyzed with either acid trehalase or amyloglycosidase for trehalose (I) and glycogen (II), respectively. The glucose produced was determined by FIA using a glucose oxidase method (Benthin et al., Anal. Chim. Acta, 1992, 261, 145). The results obtained compared well with those obtained by traditional methods and the RSD were 1.6% and 1.8% for I and II, respectively. A simple, sensitive and non-laborious enzyme-based method has been developed for determination of both trehalose and glycogen in yeast cells. The method is based on extraction of trehalose and glycogen into a 40 mM acetate buffer (pH 4.8) by mechanical disintegration of the cells in a bead mill. Subsequently, trehalose and glycogen can be hydrolyzed to glucose by the enzymes trehalase and amyloglycosidase, respectively. The formed glucose is quantified by a flow injection analyzer based on the enzyme glucose oxidase. The method gives results comparable to traditional methods but the simplicity of the analysis results in a much lower relative standard deviation. The excellent sensitivity of the glucose analyzer means that as little as 1 µg trehalose or glycogen can be determined which reduces the required sample volume. This makes the method ideal for physiological studies, e.g., of transients in continuous cultures of Saccharomyces cerevisiae. In addition, a consistent procedure has been derived for pretreatment and storage of samples.
Trehalose Glycogen Electrode Biotechnology Immobilized enzyme

"Application Of Biosensors With An Electrolyte Isolator Semiconductor Capacitor (EIS-CAP) Transducer For Process Monitoring"
Process Biochem. 1998 Volume 33, Issue 2 Pages 175-180
C. Menzel, T. Lerch, K. Schneider, R. Weidemann, C. Tollnick, G. Kretzmer, T. Scheper and K. Schüger*

Abstract: A fluoride sensitive (pF), buffer capacity insensitive electrolyte isolator semiconductor capacitor (pF-EIS-CAP) chip was used as a transducer for biosensors with co-immobilized enzymes. Hydrogen peroxide forms fluoride ions in stoichiometric amounts in the presence of p-fluoraniline and peroxidase (POD). This was measured by the pF-EIS-CAP-sensor. The combination of this sensor with various oxidases allows the measurement of the concentration. of several analytes. Sugar syrup (glucose, maltose, maltotriose and other oligosaccharide) concentrations. were analyzed with co-immobilized β-amylase, amyloglucosidase, mutarotase, glucose oxidase and peroxidase. The glucose concentration. was analyzed with co-immobilized glucose oxidase and peroxidase, the ethanol concentration. with co-immobilized alcohol oxidase and peroxidase and the phosphate concentration. by co-immobilized nucleoside phosphorylase, xanthine oxidase and peroxidase. These (Bio-pF-EIS-CAP) sensors were integrated in flow injection analysis systems. The medium components were monitored online during Saccharomyces cerevisiae, Acremonium chrysogenum and recombinant baby hamster kidney (rBHK) cell cultivations. The agreement between the off- and online concentrations. of the medium components was within 10%.
Hydrogen peroxide Glucose Maltose Maltotriose Oligosaccharides Sensor Sensor Method comparison