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Chloromuconolactone dehalogenase ClcF of actinobacteria
Authors:Inna P Solyanikova  Elena G Plotnikova  Ekaterina S Shumkova  Irina V Robota  Natalya V Prisyazhnaya
Institution:1. Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino, Moscow Region, Russia;2. Institute of Ecology and Genetics of Microorganisms, Ural Branch, Russian Academy of Sciences, Perm, Russia;3. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia;4. Pushchino State Natural Science Institute, Pushchino, Moscow Region, Russia
Abstract:This work investigated the distribution of the clcF gene in actinobacteria isolated from different ecotopes. The gene encodes chloromuconolactone dehalogenase (CMLD) ClcF, the enzyme found to date in only one representative of Gram‐positive bacteria, Rhodococcus opacus 1CP, adapted to 2‐chlorophenol (2CP). Using primers specific to the clcF gene, from the DNA matrix of rhodococcal strains closely related to species Rhodococcus wratislaviensis (P1, P12, P13, P20, G10, KT112, KT723, BO1) we obtained PCR products whose nucleotide sequences were 100% identical to that of the clcF gene from strain R. opacus 1CP. CMLDs isolated from the biomass of strains Rhodococcus spp. G10 and P1 grown on 2CP did not differ by their subunit molecular mass deduced from the known amino acid sequence of the clcF gene from the ClcF of strain R. opacus 1CP. Matrix‐assisted laser dissociation/ionization time‐of‐flight mass spectrometry showed the presence of a peak with m/z 11,194–11,196 Da both in whole cells and in protein solutions with a ClcF activity. Thus, we have first time shown the distribution of ClcF among actinobacteria isolated from geographically distant habitats.
Keywords:Chloromuconolactone dehalogenase  actinobacteria  degradation  2‐chlorophenol  3‐chlorobenzoate
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