strain A55, Stenotrophomonas sp. strain C21 and Arthrobacter sp. strain O4 are highlighted (black circles). Vertical bar represents 0.02 units of Ku-0059436 datasheet evolutionary distance. PCR detection of heavy metal determinants in genomic DNA from bacterial isolates The presence of the copA gene encoding a multi-copper oxidase in the bacterial isolates was

studied by PCR using the Coprun primers. The bacterial strains O12, A32, A55, C21 and O4 possess the copA genes. The PCR products varied between https://www.selleckchem.com/products/Fedratinib-SAR302503-TG101348.html 1000–1200 bp. The CopA protein sequences were aligned with CopA sequences belonging to Cu-resistant bacteria and were used to construct a phylogenetic tree (Figure 4). Sequence analyses indicate that the copA genes of the isolates encode multi-copper oxidases that are involved in Cu resistance but that are not associated to degradation of phenolic compounds or polymers. The CopA MAPK Inhibitor Library manufacturer protein of Sphingomonas sp. strains O12, A32 and A55 are closely related to CopA of other α-Proteobacteria, sharing high similarity (93%) with CopA from Sphingomonas sp. S17. The CopA from Stenotrophomonas sp. strain C21 belongs to the Stenotrophomonas and Xanthomonas CopA branch of the γ-Proteobacteria

and is closely related to CopA from Stenotrophomonas maltophilia R551-3 (67% similarity). The CopA of strain Arthrobacter sp. O4 is closely related to the CopA of Actinobacteria and possess a 68% similarity with CopA from Arthrobacter sp. strain FB24. Figure 4 Phylogenetic C1GALT1 tree showing the relatedness of multi-copper oxidase CopA of the bacterial isolates. The phylogenetic tree was constructed using neighbor-joining method. Values of 1000 bootstrap

replicates above 50% are given at the branching point. Sequences of CopA proteins of the bacterial isolates Sphingomonas sp. strain O12, Sphingomonas sp. strain A32, Sphingomonas sp. strain A55, Stenotrophomonas sp. strain C21 and Arthrobacter sp. strain O4 are highlighted (black circles). Other heavy metal determinants were studied by PCR using specific primers for merA (Hg2+ resistance), merB (organomercurial resistance) and chrB (CrO4 2- resistance) genes based on C. metallidurans CH34 sequences. Using these specific primers, the merA, merB and chrB genes were not detected in the five Cu-resistant bacterial strains. Detection of plasmids in bacterial isolates Sphingomonas sp. strain O12, Sphingomonas sp. strain A32, Sphingomonas sp. strain A55 and Stenotrophomonas sp. strain C21 possessed plasmids (Figure 5). Plasmids were no detected in Arthrobacter sp. strain O4. The plasmids of these four bacterial isolates contained the copA gene encoding a multi-copper oxidase (Figure 5). Figure 5 Detection of plasmids encoding copA genes in copper-resistant bacterial isolates. A. Agarose gel electrophoresis of plasmids isolated from Sphingomonas sp. strain O12 (lane 2) Sphingomonas sp. strain A32 (lane 3), Sphingomonas sp. strain A55 (lane 4) and Stenotrophomonas sp. strain C21 (lane 5). No plasmid was observed in Arthrobacter sp.