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Proceedings Paper

Investigation of the dynamics of enzymatic cytolytic processes with high-speed spectroturbidimetry
Author(s): Alexander I. Sirota; Marina N. Shatalaeva; Nikolai G. Khlebtsov
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Paper Abstract

To determine enzymatic cytolytic activities several methods, based on the change in turbidity of a suspension of microbial cells sensitive to a given enzyme, are in common use. We have developed a method based on the measurement of turbidity spectra with a high-speed turbidimeter and on the algorithms of solution of the inverse spectroturbidimetric problem. The method was evaluated using the bacterium M.lisodeicticus and the enzyme muramidase (lysozyme) as a model system. The cytolytic activity of the lysozyme was examined with a high-speed turbidimeter developed by us, which permits measuring light extinction spectra on 12 discrete wavelengths in the range of 350 to 1000 nm practically simultaneously. This enables the kinetics of change of the light extinction spectra during lysis to be investigated in more detail. In solving the inverse spectroturbidimetric problem we calculated the mean size, number and mass-volume concentrations of M. lisodeicticus in the process of lysis. The activity of the lysozyme correlated well with the rate of change of the disperse particles' mean size in the range from the second to the fifth minute of lysis. We compared the kinetics of change of the disperse particles' mean size in the process of lysis either by the pure lysozyme or by the lysozyme-containing serum. In both cases the process proceeded similarly up to the fifth minute with the activity of the serum being equivalent to the activity of the pure lysozyme in a concentration of up to 1.0 (mu) g/ml.

Paper Details

Date Published: 31 January 1995
PDF: 7 pages
Proc. SPIE 2326, Photon Transport in Highly Scattering Tissue, (31 January 1995); doi: 10.1117/12.200858
Show Author Affiliations
Alexander I. Sirota, Samara Medical Univ. (Russia)
Marina N. Shatalaeva, Samara Medical Univ. (Russia)
Nikolai G. Khlebtsov, Institute of Biochemistry and Physiology of Plants and Microorganisms (Russia)

Published in SPIE Proceedings Vol. 2326:
Photon Transport in Highly Scattering Tissue
Sigrid Avrillier; Britton Chance; Gerhard J. Mueller; Alexander V. Priezzhev; Valery V. Tuchin, Editor(s)

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