Vývoj a konstrukce biosensorů pro detekci toxických halogenovaných látek v prostředí

Bc. Šárka Bidmanová, Ph.D.

Master's thesis

Development and construction of biosensors for detection of halogenated compounds in the environment

Abstract:

Haloalkane dehalogenases (EC 3.8.1.5) are bacterial enzymes able to remove halogen from halogenated aliphatic compounds by a hydrolytic replacement. Halogenated compounds are dangerous for living organisms due to toxic, genotoxic and teratogenic effects. There is growing need for sensitive, rapid and in situ methods for monitoring of these hazardous pollutants. Whole-cell fiber optic biosensor for detection of 1,2 dichloroethane has been developed by Campbell et al. (2006). This project was focused on development of cell-free enzymatic fiber optic biosensor with the aim to increase selectivity and reproducibility of biosensors for detection of halogenated compounds. Fluoresceinamine based pH optode was prepared and effects of different factors, e.g. way of storage or amount of fluorescence dye, were investigated to attain stable signal. Developed pH optode provided linear response in the pH range from 3.0 to 6.6. Three different immobilization methods were tested with haloalkane dehalogenase LinB from Sphingobium japonicum UT26 to ensure tight proximity of enzyme and transducer: immobilization in glutaraldehyde, sol-gel and ORMOCER. Cross-linking with glutaraldehyde was selected for immobilization of LinB in biosensors and additional parameters, e.g., composition of reaction mixture and cross-linking time, were optimized. The most optimal immobilized variant achieved 62% activity of enzyme in solution. Reference: Campbell D.W., Müller C., Reardon K.F. (2006). Biotechnol. Lett. 28: 883-887.

Keywords

haloalkane dehalogenase, fluorescence fiber optic biosensor, enzyme immobilization, environmental monitoring

Language used: English

Date on which the thesis was submitted / produced: 24. 5. 2007

Thesis defence

• Date of defence: 15. 6. 2007
• Supervisor: prof. Mgr. Jiří Damborský, Dr.

Masaryk University

Faculty of Science

Master programme / field:
Biology / General Biology