Analysis of the genes encoding the MBL-fold metallohydrolase superfamily proteins of the Pseudomonas putida BS3701 petroleum component-degrading strain

Abstract

Objective. To determine whether the genes whose products are annotated as "MBL-fold metallohydrolase superfamily" are related to the proteins of the metallo-β-lactamase group. Materials and Methods. Sequences of the 7 genes annotated as "MBL-fold metallohydrolase superfamily" were analyzed using the following resources: ClustalW, IQ-TREE, iTOL. Selection of the oligonucleotides for real-time PCR was performed using the Primer-BLAST resource. The level of gene expression was assessed using real-time PCR. MIC and MBC measuring was performed using cefepime and meropenem. The double-disc method with EDTA was used to determine the presence of MBL in the strain. Results. Analysis of the nucleotide sequences of the studied genes revealed that all of them were not included in the clade containing sequences of metallo-β-lactamase. In the exponential growth phase, mRNAs corresponding to the studied proteins were found. Determination of MIC and MBC revealed a low level of resistance to antibiotics of the β-lactamase group. The phenotypic test was negative for MBL in P. putida strain BS3701. Conclusions. The investigated genes and corresponding proteins are not related to metallo-β-lactamases. They are not involved in the resistance of P. putida BS3701 to antibiotics of the metallo-β-lactamase group.

Keywords

• resistance
• metallo-β-lactamase genes
• p putida bs3701