Open AccessEvidence for more than one form of toluene cis ‐glycol dehydrogenase from Pseudomonas putida NCIB11767
Author(s) -
Jenkins Richard O.,
Stephens Gillian M.,
Dalton Howard
Publication year - 1987
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1987.tb02269.x
Subject(s) - pseudomonas putida , mutant , strain (injury) , mutagenesis , dehydrogenase , biochemistry , reversion , biology , wild type , toluene , chemistry , gene , enzyme , microbiology and biotechnology , phenotype , organic chemistry , anatomy
A number of mutants which were unable to grow on toluene were isolated after transposon mutagenesis of Pseudomonas putida NCIB11767. All of these mutants lacked functional toluene dioxygenase and our inability to isolate other classes of mutant suggested the possible functional or structural duplication of other genes involved in toluene oxidation. However, a mutant lacking toluene cis ‐glycol (TCG) dehydrogenase (strain NG1) had previously been isolated by NTG mutagenesis. This strain was apparently a double mutant since it reverted to produce two classes of revertant characterised by strains NGL and NGS. Both revertants displayed phenotypic differences from the wild‐type strain and further reversion of strain NGS to wild‐type was demonstrated. Two protein bands (A and B) possessing TCG‐dependent dehydrogenase activity were demonstrated after non‐denaturing gel electrophoresis of extracts of strain 11767. Extracts of NGL and NGS displayed only the A or B band, respectively. The TCG dehydrogenase activities of crude extracts of NGL and NGS differed in their sensitivities to inhibitors, substrate affinities and pH optima. We suggest that strain 11767 contains two forms of TCG dehydrogenase which were the products of two different genes.