Mikrobiol. Z. 2019; 81(6):97-109.
Low Molecular Weight Pseudomonas aeruginosa Bacteriocins
Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine
The aim of the work was to study Pseudomonas aeruginosa ability to produce bacteriocins with low molecular weight, establishing their nature and belonging to a certain group of antimicrobial substances. Methods. The objects of the study were substances synthesized by six Pseudomonas aeruginosa strains – highly active bacteriocin producers. Isolation of substances with low molecular weight was carried out by dialysis through a semipermeable membrane with molecular weight cut-off (MWCO) 15 kDa. The belonging of the obtained substances to bacteriocins was checked by the influence on own producer strains, the treatment with trypsin (1 mg/ml), the detection of their absorption spectrum. The investigated substances were concentrated by ammonium sulphate, and the molecular weight was determined by SDS-PAGE. Results. The results of the dialysis of P. aeruginosa lysates for 3 and 10 days showed that the studied cultures are able to produce low-molecular substances that penetrate through the pores of dialysis membrane with MWCO 15 kDa. The duration of penetration and activity of these substances for each strain were different. But in all cases, dialysates of the 1st day were characterized by the maximum activity, and then the tendency to decrease these parameters in subsequent periods of sampling was observed. The detected substances did not penetrate through the dialysis membrane with MWCO 6-8 kDa, did not affect their own producer strains, and lost activity after treatment with trypsin (1 mg/ml). The peaks of their absorption were 205-210 nm, which is characteristic of peptide bonds of proteins. The received data allowed characterizing the detected substances as low molecular weight bacteriocins. The results of SDS-PAGE revealed a single peptide with molecular weight of 9 kDa. Conclusions. The investigated P. aeruginosa strains were characterized by production of low molecular weight microcin II-like bacteriocins with molecular weight about 9 kDa. These substances may be associated with S-type pycins, which are also synthesized by the same strains.
Keywords: low molecular weight pyocins, microcin II-like bacteriocins, Pseudomonas aeruginosa, dialysis purification, absorption peak 205-210 nm.
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