Electrolysis: processes – compositions used therein – and methods – Electrolytic analysis or testing – Involving enzyme or micro-organism
Reexamination Certificate
2000-10-02
2003-01-28
Tung, T. (Department: 1743)
Electrolysis: processes, compositions used therein, and methods
Electrolytic analysis or testing
Involving enzyme or micro-organism
C204S403100, C204S403140
Reexamination Certificate
active
06511592
ABSTRACT:
Pentachlorophenol (PCP) and other polyhalogenated phenols constitute a major group of environmental pollutants. These compounds are widely used as wood preservatives, fungicides, insecticides and bactericides. PCP is a biocide and used primarily for wood preservation. It is ubiquitous in the environment and considered a priority pollutant by the U.S. Environmental Protection Agency. The Dutch proposal for soil contamination limit is ca. 5 mg/kg dry weight. Management of pollution byPCP (or indeed by any man-made chemical) of the environment involves monitoring and where necessary, remediation. For priority pollutants such as PCP, it is therefore highly desirable to have an easy, point-of-use, real-time sensor—the development of such a sensor is the subject of this application.
BACKGROUND
The monooxygenase enzyme pentachlorophenol hydroxylase (PCPH) is responsible for the first oxidation step in the pentachlorophenol degradation pathway in Flavobacterium sp. (ATCC 39723). PCPH catalyses the oxidation of pentachlorophenol (PCP) to 2,3,5,6-tetrachloro-p-hydroquinone (TeCH), a reaction which requires two equivalents of NADPH and one molecule of dioxygen (Scheme 1). The gene encoding this 65 kDa flavoenzyme has been cloned and sequenced, and the enzyme has been over-expressed in
E. coli.
The substrate range of PCPH has been investigated. It was found that the enzyme has an absolute requirement for the phenol group and also a substituent at the ortho position to the phenol. Thus, benzene and halobenzenes are not substrates, nor does PCPH attack 3,5-dichlorophenol although 2,3-dichlorophenol is a poor substrate.
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Abstract of Xun et al. (“Confirmation of oxdidative dehalogenation of pentachlorophenol by a Flavobacterium pentachlorophenol hydroxylase”, J. Bacteriol. Sep. 1992, 5745-5747, vol. 174, No. 17).*
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K. B. Male et al., “Optimization and Characterization of a Flow Injection Electrochemical System for Pentachlorophenol Assay,”Anal Chem.,vol. 70, pp. 4134-4139 (Oct. 1998).
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Hill Hugh Allen Oliver
Jones Jonathan Peter
Wong Luet-Lok
Xie Weihong
ISIS Innovation Limited
Lahive & Cockfield LLP
Laurentano Anthony A.
Lauro Peter C.
Noguerola Alex
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