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Rapid evaluation of biocidal activity using a transposon‐encoded catechol 2,3‐dioxygenase from Pseudomonas putida
Author(s) -
Edghill L. A.,
Russell A. D.,
Day M. J.,
Furr J. R.
Publication year - 1999
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1046/j.1365-2672.1999.00801.x
Subject(s) - catechol , pseudomonas putida , chemistry , phenol , chromatography , pseudomonas , cetylpyridinium chloride , biochemistry , microbiology and biotechnology , bacteria , nuclear chemistry , enzyme , biology , organic chemistry , pulmonary surfactant , genetics
Pseudomonas putida (UWC1), containing a genetically‐engineered plasmid (pQM899), that encodes for the production of catechol 2,3‐dioxygenase (C230), was used as a potential means of rapidly estimating bactericidal activity of chlorhexidine diacetate (CHA), phenol, cetylpyridinium chloride (CPC) and phenylmercuric nitrate (PMN). Enzyme C230 converts catechol to 2‐hydroxymuconic semialdehyde (2‐HMS), which is yellow in colour, via a meta cleavage pathway. Ideal conditions for production and measurement spectrophotometrically of 2‐HMS were determined. However, the correlation between this method and viable plate counts was not sufficiently accurate to enable 2‐HMS production to provide a sufficiently sensitive determination of biocidal activity. An alternative method, synchronous scanning fluorimetry, in which the decrease in catechol concentration was measured under standardized conditions, provided a good dose–response histogram for all the biocides tested. Although, in comparison with plate counts, there was an underestimation of the bactericidal effects of phenol and PMN, the results of this study suggest that this method has potential in determining the bactericidal efficacy of agents such as CHA and CPC