Development of an enzymatic fiber-optic biosensor for detection of halogenated hydrocarbons.

BIDMANOVÁ, Šárka, Radka CHALOUPKOVÁ, Jiří DAMBORSKÝ and Zbyněk PROKOP. Development of an enzymatic fiber-optic biosensor for detection of halogenated hydrocarbons. Analytical and Bioanalytical Chemistry. Springer-Verlag, 2010, vol. 10, No 1007, p. 1891-1898. ISSN 1618-2642.

Basic information

• Original name: Development of an enzymatic fiber-optic biosensor for detection of halogenated hydrocarbons.
• Authors: BIDMANOVÁ, Šárka (203 Czech Republic, belonging to the institution), Radka CHALOUPKOVÁ (203 Czech Republic, belonging to the institution), Jiří DAMBORSKÝ (203 Czech Republic, guarantor, belonging to the institution) and Zbyněk PROKOP (203 Czech Republic, belonging to the institution).
• Edition: Analytical and Bioanalytical Chemistry, Springer-Verlag, 2010, 1618-2642.

Other information

• Original language: English
• Type of outcome: Article in a journal
• Field of Study: 10600 1.6 Biological sciences
• Country of publisher: Germany
• Confidentiality degree: is not subject to a state or trade secret
• WWW: URL
• Impact factor: Impact factor: 3.841
• RIV identification code: RIV/00216224:14310/10:00040599
• Organization unit: Faculty of Science
• UT WoS: 000283244500008
• Keywords (in Czech): Enzyme-based biosensor; Haloalkán dehalogenáza; Halogenované uhlovodíky; Optické biosensory; pH indikátory
• Keywords in English: Enzyme-based biosensor; Haloalkane dehalogenase; Halogenated hydrocarbons; Optical biosensor; pH indicator

Abstract

An enzyme-based biosensor was developed by co-immobilization of purified enzyme haloalkane dehalogenase (EC 3.8.1.5) and a fluorescence pH indicator on the tip of an optical fiber. Haloalkane dehalogenase catalyzes hydrolytic dehalogenation of halogenated aliphatic hydrocarbons, which is accompanied by a pH change influencing the fluorescence of the indicator. The pH sensitivity of several fluorescent dyes was evaluated. The selected indicator 5(6)-carboxyfluorescein was conjugated with bovine serum albumin and its reaction was tested under different immobilization conditions. The biosensor was prepared by cross-linking of the conjugate in tandem with haloalkane dehalogenase using glutaraldehyde vapor. The biosensor, stored for 24 h in 50 mM phosphate buffer (pH 7.5) prior to measurement, was used after 15 min of equilibration, the halogenated compound was added, and the response was monitored for 30 min. Calibration of the biosensor with 1,2-dibromoethane and 3-chloro-2-(chloromethyl)- 1-propene showed an excellent linear dependence, with detection limits of 0.133 and 0.014 mM, respectively. This biosensor provides a new tool for continuous in situ monitoring of halogenated environmental pollutants.

Links

• ED0001/01/01, research and development project: Name: CETOCOEN
• IAA401630901, research and development project: Name: Evoluce substrátové specifity u enzymů aktivních s xenobiotickými látkami

Investor: Academy of Sciences of the Czech Republic, Evolution of substrate specificity in enzymes acting on xenobiotic compounds

• MSM0021622412, plan (intention): Name: Interakce mezi chemickými látkami, prostředím a biologickými systémy a jejich důsledky na globální, regionální a lokální úrovni (INCHEMBIOL) (Acronym: INCHEMBIOL)

Investor: Ministry of Education, Youth and Sports of the CR, Interactions among the chemicals, environment and biological systems and their consequences on the global, regional and local scales (INCHEMBIOL)