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. 2023 Dec 19;13(1):10.
doi: 10.3390/foods13010010.

Simple Purification and Antimicrobial Properties of Bacteriocin-like Inhibitory Substance from Bacillus Species for the Biopreservation of Cheese

Jong-Hui Kim  1 , Eun-Seon Lee  1 , Bu-Min Kim  1 , Jun-Sang Ham  1 , Mi-Hwa Oh  1
Affiliations

Simple Purification and Antimicrobial Properties of Bacteriocin-like Inhibitory Substance from Bacillus Species for the Biopreservation of Cheese

Jong-Hui Kim et al. Foods. .

Abstract

Bacteriocins may be used as natural preservatives and antibiotic substitutes in various foods. However, the multistep purification process of bacteriocins results in high production costs, which is an obstacle to their commercial use and consumer accessibility. In this study, a bacteriocin-like inhibitory substance (BLIS) from Bacillus spp. isolated from Korean fermented foods was partially purified using the aqueous two-phase system (ATPS). The maximum activity of the BLIS was achieved for ATPS composed of PEG 1000 (15% [w/w])/ammonium sulfate (20% [w/w])/sodium chloride (2% [w/w]), which caused BLIS activity to increase by 3 times with a 99% recovery rate. In particular, B. amyloliquefaciens Y138-6 BLIS exhibited broad antibacterial activity, high resistance to acid-base stress, and excellent thermal stability. This antibacterial substance inhibited the growth of aerobic bacteria and fungi on the walls of cheese and ripening rooms. These antibacterial properties have been shown to increase food safety and have the potential for use as biopreservatives. Moreover, considering that the execution of the ATPS requires only salts and PEG, it is a simple, environmentally friendly, and cost-effective process and may have industrial applications in the recovery of BLIS from fermentation broth.

Keywords: Bacillus; aqueous two-phase system; bacteriocin-like inhibitory substance.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Distribution of Bacillus species diversity as determined in different Korean fermented foods. Kimchi, n = 111; pickles, n = 96; sauces, n = 79; and salted fish, n = 9.
Figure 2
Figure 2
(a) Optimal temperature and (b) pH for BLIS production in Bacillus isolates in tryptic soy broth medium.
Figure 3
Figure 3
Cytotoxicity of partially purified BLIS from Bacillus isolates on Caco-2 cells in vitro. Data were expressed as the mean ± standard deviation (n = 5). N.S. = no significance when compared with the negative control group (DW); Student’s t-test. *** p < 0.001.
Figure 4
Figure 4
Changes in the number of (a) aerobic bacteria and (b) fungi on the surface of Gouda cheese and the number of (c) aerobic bacteria and (d) fungi in the cheese-ripening room walls after exposure to partially purified Y138-6 BLIS.
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