Ultrasonic velocity assay of extracellular invertase in living yeasts.
The use of a low-intensity ultrasonic technique (noninvasive, nondestructive, on-line, and able to assess opaque samples) to monitor the kinetics of invertase hydrolysis is presented. Adiabatic compressibility has been shown to be sensitive to sugar species: ultrasonic velocity increasing as saccharose is transformed into glucose and fructose. The influence of initial sucrose mass concentration (2-60%), temperature (25-55 degrees C), pH (3.5-6.5), and number of microorganisms (10(5)-10(9) yeasts/ml) on the reaction rate, catalyzed by the extracellular invertases of intact Saccharomyces cerevisiae cells, has been measured. The results were proven to be in strict agreement with the optimal kinetic parameters of the enzyme. Ultrasonic velocity variations are explained in terms of changes of the solute concentrations in the mixture water-saccharose-glucose/fructose and calculated from the velocity of ultrasound in the corresponding pure sugar solutions. A linear relationship between the initial rate of ultrasonic velocity and the number of yeasts (enzymes) is pointed out.
Departamento de Señales, Sistemas y Tecnologías Ultrasónicos, Instituto de Acústica (CSIC), Serrano 144, 28006 Madrid, Spain. blopa@ia.cetef.csic.es
Pablo
P
Resa
Luis
L
Elvira
Carlos
C
Sierra
Francisco Montero de
FM
Espinosa
D005110:Extracellular Space / Q000201:enzymology* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000378:metabolism; D013696:Temperature; D014465:Ultrasonics; D043324:beta-Fructofuranosidase / Q000378:metabolism*
D043324:beta-Fructofuranosidase