A New Method for Determination of Enzyme Reaction and Activity of Lysozyme with UV-Spectrophotometer

UV-분광광도계를 이용한 새로운 Lysozyme의 효소반응 및 활성측정법 연구

  • Kim, Woon-Soo (Department of Chemical Engineering, Kyunghee University) ;
  • Kim, Yong-Wook (Department of Chemical Engineering, Kyunghee University) ;
  • Kim, Woo-Sik (Institute of Materials Science & Technology, Kyunghee University)
  • 김운수 (경희대학교 화학공학과) ;
  • 김용욱 (경희대학교 화학공학과) ;
  • 김우식 (경희대학교 재료과학연구소)
  • Received : 1998.05.07
  • Accepted : 1998.07.30
  • Published : 1998.11.10

Abstract

A simple and new experimental method for determination of lysozyme-M. lysodeikticus cell lysis reaction and lysozyme activity was suggested using Beer's law. The UV transmittance of the solution changed with the concentration of M. lysodeikticus and the relationship between the UV transmittance and M. lysodeikticus cell concentration followed Beer's Law. In addition, it was experimentally proven that the UV transmittance of the solution was not influenced by the lysozyme concentration and product of the lysis reaction. During the lysozyme-M. lysodeikticus cell lysis reaction, thus, M. lysodeikticus cell concentration in the solution could be measured in-situ by UV-spectrophotometer. By using these experimental data, kinetic Parameters of the Michaelis-Menten equation for the lysozyme-M. lysodeikticus cell 1ysis reaction was simply determined The maximum reaction rate constant ($k_3$) and Michaelis-Menten constants were $0.1734sec^{-1}$ and $9.83{\times}10^{-6}M$ respectively. The activity of the lysozyme could also be obtained with this experiment because the lysis reaction rate of the 1ysozyme depended on its activity.

Acknowledgement

Supported by : 한국과학재단

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