The gene of sp. strain LB400 has previously been shown to be located within the locus, which specifies the degradation of biphenyl (BP) and chlorobiphenyls, and to encode a glutathione -transferase (GST) which accepts 1-chloro-2,4-dinitrobenzene (CDNB) as substrate. The specific physiological role of this gene is not known. It is now shown that the gene is expressed in the parental organism and that GST activity is induced more than 20-fold by growth of the strain on BP relative to succinate when these compounds serve as sole carbon source. Approximately the same induction factor was observed for 2,3-dihydroxybiphenyl 1,2-dioxygenase activity, which is encoded by the 5′-adjacent gene. This suggests that the expression of is coregulated with the expression of genes responsible for the catabolism of BP. A probe detected only a single copy of the gene in strain LB400. A spontaneous BP mutant of the organism neither gave a signal with the probe nor showed CDNB-accepting GST activity, suggesting that this activity is solely encoded by . Complementation of the mutant with a gene cluster devoid of restored the ability to grow on BP, indicating that is not essential for utilization of this carbon source. BphK activity proved to be almost unaffected by up to 100-fold differences in proton concentration or ionic strength. The enzyme showed a narrow range with respect to a variety of widely used electrophilic GST substrates, accepting only CDNB. A number of established laboratory strains as well as novel isolates able to grow on BP as sole carbon and energy source were examined for BphK activity and the presence of a analogue. CDNB assays, probe hybridizations and PCR showed that several, but not all, BP degraders possess this type of GST activity and/or a closely related gene. In all bacteria showing BphK activity, this was induced by growth on BP as sole carbon source, although activity levels differed by up to 10-fold after growth on BP and by up to 60-fold after growth on succinate. This resulted in a variation of induction factors between 2 and 30. In the majority of bacteria examined, the gene appeared to be part of LB400-like gene clusters. DNA sequencing revealed almost complete identity of genes from five different gene clusters. These results suggest that genes, although not essential, fulfil a strain-specific function related to the utilization of BPs by their host organisms. The usefulness of BphK as a reporter enzyme for monitoring the expression of catabolic pathways is discussed.


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