Purification of bacteriocins by chromatographic methods

Surovtsev, V. I.; Borzenkov, V. M.; Levchuk, Vladimir P.

doi:10.1134/s0003683815090069

Cited by 6 publications

(5 citation statements)

References 8 publications

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“…Thus, it is interesting to conduct a comparative study on the stability between crude, partially purified, and purified forms of bacteriocin from P. acidilactici kp10 to further characterise the compound prior to commercial applications. Purified or partially purified bacteriocins of LAB origins can be obtained from various techniques of purification based on their affinity to organic solvents and solubility variations in concentrated salt solution and pH [ 58 – 62 ].…”

Section: Discussionmentioning

confidence: 99%

Stability of Bacteriocin-Like Inhibitory Substance (BLIS) Produced byPediococcus acidilacticikp10 at Different Extreme Conditions

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Tan

et al. 2018

BioMed Research International

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Nowadays, bacteriocin industry has substantially grown replacing the role of chemical preservatives in enhancing shelf-life and safety of food. The progress in bacteriocin study has been supported by the emerging of consumer demand on the applications of natural food preservatives. Since food is a complex ecosystem, the characteristics of bacteriocin determine the effectiveness of their incorporation into the food products. Among four commercial media (M17 broth, MRS broth, tryptic soy broth, and nutrient broth) tested, the highest growth of Pediococcus acidilactici kp10 and bacteriocin-like-inhibitory substance (BLIS) production were obtained in the cultivation with M17. BLIS production was found to be a growth associated process where the production was increased concomitantly with the growth of producing strain, P. acidilactici kp10. The antimicrobial property of BLIS against three indicator microorganisms (Listeria monocytogenes, Escherichia coli, and Staphylococcus aureus) remained stable upon heating at 100°C but not detectable at 121°C. The BLIS activity was also observed to be stable and active at a wide pH range (pH 2 to pH 7). The BLIS activity remained constant at -20°C and -80°C for 1 month of storage. However, the activity dropped after 3 and 6 months of storage at 4°C, -20°C, and -80°C with more than 80% reduction. The ability of bacteriocin from P. acidilactici kp10 to inhibit food-borne pathogens while remaining stable and active at extreme pH and temperature is of potential interest for future applications in food preservatives.

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Section: Discussionmentioning

confidence: 99%

Stability of Bacteriocin-Like Inhibitory Substance (BLIS) Produced byPediococcus acidilacticikp10 at Different Extreme Conditions

Sidek

Halim

Tan

et al. 2018

BioMed Research International

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“…By a combination of precipitation with ammonium sulfate at 60% saturation followed by hydrophobic chromatography on SepPak C18 with elution with a stepwise gradient of isopropanol in 25 mM potassium phosphate buffer at pH 6.5, fractions of bacteriocins from P. acidilactici ST3522BG, P. pentosaceus ST3633BG, and E. faecium ST7319ea were obtained predominantly in the 60% and 80% isopropanol fractions. The combination of ammonium sulfate precipitation and hydrophobic chromatography on SpePakC 18 can be considered a highly successful approach for the partial purification of bacteriocins produced by LAB and was applied formerly in purification protocols by Todorov et al [ 22 ], Metivier et al [ 41 ], Bughaloo-Vial et al [ 42 ], Song et al [ 43 ], and Surovtsev et al [ 44 ]. By nature, all bacteriocins are proteinaceous, and ammonium sulfate precipitation combined with the level of saturation gives excellent results for the preliminary extraction of these proteinaceous molecules with a specific range of molecular sizes [ 45 ].…”

Section: Resultsmentioning

confidence: 99%

“…The use of hydrophobic SepPakC 18 columns exploits the hydrophobic character of most bacteriocins. By further application of an isopropanol gradient, these antimicrobial peptides can be eluted at a specific point [ 44 ]. These approaches were previously applied in the partial purification of several bacteriocins [ 22 , 41 , 42 , 43 , 44 ].…”

Section: Resultsmentioning

confidence: 99%

“…By further application of an isopropanol gradient, these antimicrobial peptides can be eluted at a specific point [ 44 ]. These approaches were previously applied in the partial purification of several bacteriocins [ 22 , 41 , 42 , 43 , 44 ]. The obtained active fractions can be applied for the purpose of analytical experiments or can be subjected to further purification, depending on the objective of the research.…”

Section: Resultsmentioning

confidence: 99%

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Combined Action of Antibiotics and Bacteriocins against Vancomycin-Resistant Enterococci

et al. 2022

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Antibiotics have been one of the most important discoveries in the area of applied medical microbiology; however, as a result of various factors, we are currently facing a dramatic and relatively dangerous increase in the number of cases of antibiotic resistance, and the need for new types of antimicrobials continues to grow. New approaches are needed to combat antibiotic-resistant pathogens. Bacteriocins, as part of the group of antimicrobial peptides, can be considered as alternatives and/or complements to known antibiotics. Their narrow spectra of activity can be explored for the control of various pathogens, such as vancomycin-resistant enterococci (VRE), as single therapies or in combination with known antibiotics. In the present study, we isolated bacteriocins from different lactic acid bacteria (LAB) strains, including Enterococcus and Pediococcus, and explored the possible synergistic inhibition of growth by bacteriocins and vancomycin. It was observed in the growth dynamics with previously selected VRE strains that the bacteriocins had a high specificity and a promising inhibitory effect against the VRE strains, and these results were validated by a propidium iodide viability test using flow cytometry. The data obtained indicate that the selected bacteriocins can be used to control VRE in the food industry or even as an alternative treatment to combat infections with antibiotic-resistant bacteria.

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“…For the partial purification of the produced by studied LAB antimicrobial proteins, recommendations from Bughaloo-Vial et al [ 31 ], Song et al [ 32 ], and Surovtsev et al [ 33 ] were followed. Two LAB strains selected in the screening tests for activity against C. albicans 1281 ( E. mundtii CRL35 and E. faecium ST88Ch) were cultured in 500 mL MRS broth for 24 h at 37 °C.…”

Section: Methodsmentioning

confidence: 99%

In Vitro Anti-Candida albicans Mode of Action of Enterococcus mundtii and Enterococcus faecium

et al. 2023

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Candida albicans is an important vaginosis causative agent, affecting several women worldwide each year. This study reports on two strains of lactic acid bacteria (Enterococcus mundtii CRL35 and Enterococcus faecium ST88Ch) expressing bacteriocin-like inhibitor substances (BLIS) active against C. albicans 1281. Both strains were γ-hemolytic and not affected by numerous antibiotics, contraceptives, and commercial drugs, suggesting safety for human use. The recorded antimicrobial activity of semi-purified BLIS was 25,600 AU/mL for E. mundtii CRL35 and 800 AU/mL for E. faecium ST88Ch. Treatment of BLIS with 1 mg/mL proteinase K resulted in complete loss of antimicrobial activity against Listeria monocytogenes ATCC 15313 and partial loss of activity against C. albicans 1281. The killing effect of the semi-purified BLIS on cell suspensions of C. albicans 1281 after 9 h of contact was dose-dependent: for E. mundtii CRL35, 400 AU/mL to 25,600 AU/mL caused 63.61% to 79.35% lysis, while for E. faecium ST88Ch, 200 AU/mL to 800 AU/mL caused 29.32% to 31.25% cell lysis. The effects of temperature, pH, and presence of the contraceptive Nordette-28 on the adsorption levels of the BLIS to C. albicans 1281 were also evaluated. Nordette-28 (10% or 20%) promoted increased adsorption of both studied BLIS to the cells of C. albicans 1281 at pH 5.0, while a minor effect was observed at pH 3.0. Different levels of aggregation between C. albicans 1281 and E. mundtii CRL35 or E. faecium ST88Ch were recorded, and optimal adsorption levels were recorded at 37 °C. Appropriate BLIS-producing strains can effectively contribute to the equilibrium of vaginal microbial status quo and reduce negative consequences from the development of C. albicans infections.

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Purification of bacteriocins by chromatographic methods

Cited by 6 publications

References 8 publications

Stability of Bacteriocin-Like Inhibitory Substance (BLIS) Produced byPediococcus acidilacticikp10 at Different Extreme Conditions

Stability of Bacteriocin-Like Inhibitory Substance (BLIS) Produced byPediococcus acidilacticikp10 at Different Extreme Conditions

Combined Action of Antibiotics and Bacteriocins against Vancomycin-Resistant Enterococci

In Vitro Anti-Candida albicans Mode of Action of Enterococcus mundtii and Enterococcus faecium

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