ISOLATION AND CHARACTERIZATION OF MICROORGANISMS THAT DEGRADE 4-CHLOROBIPHENYL TO 4-CHLOROBENZOIC ACID

Abstract

We designed a simple method that assesses the ability of aerobic microorganisms in soils to degrade 4-chlorobiphenyl (4-CB) to 4-chlorobenzoic acid. We screened bacteria in 46 soil samples collected from fields, paddy fields and gardens for strains that grew on biphenyl and degraded 4-CB. Sixty-five strains that utilized biphenyl were isolated from 21(46%) of 46 soil samples, a high frequency. A simple screening method that used high-pressure liquid chromatography showed that 62 (95%) of the 65 strains that used biphenyl also degraded 4-CB. The results of metabolite analysis indicated that each isolate, without exception, produced 4-chlorobenzoic acid in a, culture medium with a transitory yellow tinge. Metabolites of 4-CB were further investigated in an assay that used resting cells of Bacillus sp. strain TH 1171. The yellow metabolite was found to be 2-hydroxy-6-oxo-6-(4′-chlorophenyl)hexa-2, 4-dienoic acid from the mass spectrum of its trimethylsilyl derivative. We examined the ability of the resting cells of strain TH 1171 incubated for 24h with a mixture of PCB congeners to degrade PCB by gas chromatography-mass spectrometry. Congeners 2-, 3-, and 4-chlorobiphenyl completely disappeared during incubation. 2, 2′-, 2, 3′-, and 2, 4′-Dichlorobiphenyl and 2, 4-, 2, 5-, and 2, 6-dichlorobiphenyl were transformed into mono- and dichlorobenzoic acids, respectively. The resulting degradation of 2, 2′-, 2, 3′-, and 2, 4′-dichlorobiphenyl indicates that strain TH 1171 degrades chlorophenyl moieties of the PCB congeners. A possible biodegradation pathway of 4-CB via 2, 3-dihydroxy-4′-chlorobiphenyl is proposed.