Home HOME General Physiology and Biophysics 2020 General Physiology and Biophysics Vol.39, No.4, p.301–318, 2020

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Founded: 1982
ISSN  1338-4325 (online)

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General Physiology and Biophysics Vol.39, No.4, p.301–318, 2020

Title: A new type strain of Pseudomonas putida NM-CH-I15-I isolated from a nickel-contaminated soil in southwest Slovakia harbouring the czcA-NM15I resistance determinant
Author: Nikola Mihalíková, Janka Harichová, Marcel Zámocký, Anna Kamlárová, Peter Ferianc

Abstract: A soil bacterium MR-CH-I15-I was on the base of phylogenetic analysis of almost the whole (1,492 bp) 16S rRNA sequence and sequences of selected 9 marker genes identified as Pseudomonas putida strain NM-CH-I15-I. The bacterium exhibited typical morphological features and biochemical properties for this species, the highest resistance to nickel and copper and multidrug resistance to different antibiotic groups. In addition, the whole czcA-NM15I heavy-metal resistance gene sequence (3,126 bp, 1,042 amino acids, MW 112, 138 Da) was obtained and on the base of phylogenetic analysis was assigned to CzcA protein from Pseudomonas reidholzensis with 93% similarity. This gene was significantly induced mainly by the addition of zinc, cadmium and cobalt and in a lesser extent of nickel. Furthermore, an increased expression of the CzcA-NM15I protein was confirmed by immunoblot analysis after heterologous expression of the czcA-NM15I synthetic variant gene in E. coli BL21 (DE3). Finally, the location of amino acids (R83, R673, D402, D408, D619, E415, E568) in the homology model of the CzcA-NM15I protein suggested that these amino acids may play an important role in the transport of cations such as cobalt, zinc or cadmium. This soil bacterium can represent a new type strain of P. putida NM-CH-I15-I.

Keywords: Pseudomonas putida type strain — Molecular marker genes — czcA resistance determinant — RT-qPCR — Molecular modelling
Published online: 17-Aug-2020
Year: 2020, Volume: 39, Issue: 4 Page From: 301, Page To: 318

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