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SCREENING OF LACTOBACILLUS STRAINS FOR KNOWN AND SUGGESTED BACTERIOCIN GENES BY QUANTITATIVE POLYMERASE CHAIN REACTION
Abstract
The antibiotic resistance is one of biosphere scale problem created by not only the medicine, but veterinary, food industry and animal husbandry. Bacteriocins are defined as ribosomally synthesized peptides with narrow or in some cases broad antimicrobial activity producing by various bacteria, especially against closely-related species. Among bacteriocin producers food-grade lactic acid bacteria (LAB) provide an increase interest due to their safety, preservative ability and potential probiotic properties. Unlike traditional antibiotics, bacteriocins are active at nanomolar concentration, have less toxicity and completely metabolized in the human body, that makes the use of bacteriocins more preferable in some cases, including treatment of infections caused by multidrug-resistant bacteria, food processing, agricultural production. The aim of this study was to identify bacteriocins structural genes in Lactobacillus species based on known and predicted bacteriocins gene sequences in the NCBI GenBank database using quantitative polymerase chain reaction (qPCR). Twenty primer pairs were designed for identification chromosome or plasmid encoded bacteriocins of Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus sakei and Lactobacillus salivarius strains from the Russian National Collection of Industrial Microorganisms (VKPM). Our research was focused on the detection of class II bacteriocins such as plantaricins of L. plantarum, sakacins of L. sakei, acidocins of L. acidophilus and a number of III class bacteriocins such as helveticin J and its homologs. qPCR technique was optimized for identification of bacteriocin genes. Structural genes coding for class II (heat-stable unmodified bacteriocins) and III (heat-labile proteins) bacteriocins of Lactobacillus were successfully amplified, which was also confirmed by melting curves and agarose gel electrophoresis. The results suggest that bacteriocinogenic potential of probiotic Lactobacillus strains from VKPM has been confirmed.
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