• Title/Summary/Keyword: Bacteriocin activity

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Heterologous Expression of Human $\beta$-Defensin-1 in Bacteriocin-Producing Laetoeoeeus lactis

  • CHOI HAK JONG;SEO MYUNG JI;LEE JUNG CHOUL;CHEIGH CHAN ICK;PARK HOON;AHN CHEOL;PYUN YU RYANG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.330-336
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    • 2005
  • Lactococcus lactis A164 is a nisin Z-producing strain isolated from kimchi. Its antimicrobial spectrum has been found to be active against most Gram-positive bacteria tested, yet inactive against Gram-negative bacteria [3]. Accordingly, to overcome this drawback, the current study attempted to express human $\beta$-defensin-l (hBD-l), which kills both Gram-positive and Gram-negative bacteria in L. lactis AI64. When the hBD-l cDNA was introduced using a nisin Z-controlled expression cassette, the L. lactis A164 transformants grew very poorly, due to the bactericidal effect of the expressed hBD-l against the transformants. Therefore, a gene fusion system was designed to reduce the toxicity of the expressed heterologous protein against the host cells. As such, the hBD-l gene was fused to the DsbC- Tag of pET -40b(+), then introduced to L. lactis A 164. The transformants expressed an intracellular 35.6-kDa DsbC-hBD-l fusion protein that exhibited slight activity against the host cells, yet not enough to strongly inhibit the cell growth. To obtain the recombinant hBD-l, the DsbC-hBD-l fusion protein was purified by nickel-affinity column chromatography, and the DsbC-Tag removed by cleaving with enterokinase. The cleaved mature hBD-l exhibited strong bactericidal activity against E. coli JM109, indicating that the recombinant L. lactis A 164 produced a biologically active hBD-I. In addition, the recombinant L. lactis A 164 was also found to produce the same level of nisin Z as the wild-type.

Characterization of Antibacterial Compounds from Bacillus polyfermenticus CJ6 and Its Growth Inhibition Effect on Food-Borne Pathogens (Bacillus polyfermenticus CJ6가 생산하는 항세균 물질의 특성 및 병원성 식중독 미생물의 성장 억제 효과)

  • Jung, Ji-Hye;Chang, Hae-Choon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.6
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    • pp.903-911
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    • 2011
  • In this study, Bacillus polyfermenticus CJ6 harboring antibacterial activity was isolated from meju. The antibacterial activity of Bacillus polyfermenticus CJ6 was stable in the pH range of 3.0~9.0, but it disappeared after culture at $70^{\circ}C$ for 24 hr. Antibacterial activity was inactivated by proteinase K, protease, and ${\alpha}$-chymotrypsin, indicating its proteinaceous nature. The growth inhibitory effects of B. polyfermenticus CJ6 culture on food-borne pathogens such as Staphylococcus aureus, Salmonella Typhi, Listeria monocytogenes, and Escherichia coli O157:H7 were examined in this study. Approximately 6~6.2 log CFU/mL of each pathogen was co-cultured with B. polyfermenticus CJ6 in a 50 mL culture volume for 24 hr. Growth of S. aureus and L. monocytogenes was completely inhibited after 3 hr of incubation. Growth of S. Typhi and E. coli O157:H7 was also completely inhibited after 6 hr of incubation. The antibacterial compounds from B. polyfermenticus CJ6 were purified by solid phase extraction (C18 Sep-pak cartridge), recycling preparative HPLC, and analytical HPLC. Ultra-high performance liquid chromatography and electrospray ionization tandem mass spectrometry analysis were used to identify the purified antibacterial compounds, which were confirmed to be five peptides (757.4153 Da, 750.3444 Da, 1024.5282 Da, 1123.6083 Da, and 1617.8170 Da).

Properties of Antimicrobial Substances Produced by Bacillus amyloliquefaciens CJW15 and Bacillus amyloliquefaciens SSD8 (Bacillus amyloliquefaciens CJW15와 SSD8이 만드는 항균물질들의 특성)

  • Liu, Xiaoming;Shim, Jae Min;Yao, Zhuang;Lee, Jae Yong;Lee, Kang Wook;Kim, Hyun-Jin;Ham, Kyung-Sik;Kim, Jeong Hwan
    • Microbiology and Biotechnology Letters
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    • v.44 no.1
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    • pp.9-18
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    • 2016
  • Two Bacillus strains, CJW15 and SSD8, with strong antibacterial activities were isolated from cheonggukjang. Both were identified as B. amyloliquefaciens strains after gene sequencing of rRNA and recA. CJW15 strongly inhibited the growth of B. cereus (ATCC14579), Listeria monocytogenes (ATCC19111), and Lactococcus lactis (ATCC11454). In comparison, SSD8 inhibited the growth of B. cereus (ATCC14579) and Enterococcus faecium (ATCC19953). The antibacterial activities of the two strains were not affected when exposed to a temperature of $100^{\circ}C$ for 15 min and were quite stable in acidic (pH 3) and alkaline (pH 12) pH conditions. Enzymatic treatments (trypsin, pepsin, proteinase K, and protease) had no effect on the activity of CJW15, but reduced the activity of SSD8 by half. Both isolates possess genes involved in the synthesis of lipopeptides (e.g. surfactin, fengycin, iturin, and iturin A), and genes encoding subtilin, a bacteriocin. Moreover, both isolates have fibrinolytic activities as well.

Application of Functional Microbial Strains Isolated from Traditional Rice Wine in Korea (막걸리 유래 미생물의 활용을 위한 연구 동향)

  • Lee, Youngsuk;Seol, Jeongman;Jeong, Deokyeol;Kim, Soo Rin
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.229-235
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    • 2016
  • Korea has a long history of brewing traditional rice wine using a authentic starter culture called nuruk, which contains natural microbial flora. Because rice wine is consumed fresh without filtration, its viable cells contribute to the biological activities of the wine. In numerous studies, microbial strains isolated from rice wine have been screened for their functionalities, which were mainly probiotic properties and antimicrobial activities. Indeed, some lactic acid bacteria (LAB) were confirmed to have strong probiotic activities as well as other health-promoting effects. Moreover, some of the isolated probiotic strains produced functional compounds, such as exopolysaccharides and γ-aminobutyric acid. For antimicrobial activities, some LAB and yeast strains were identified to produce bacteriocins and killer toxins, respectively, with significantly broad spectrum of antimicrobial activity. These functional strains originating from traditional rice wine and their metabolites can be used directly for the production of value-added food products.

Potential Application of Bacteriocin Produced from Lactic Acid Bacteria

  • El Issaoui, Kaoutar;Senhaji, Nadia Skali;Zinebi, Sanae;Zahli, Rajae;Haoujar, Imane;Amajoud, Nadia;Abrini, Jamal;Khay, El Ouardy
    • Microbiology and Biotechnology Letters
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    • v.48 no.3
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    • pp.237-251
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    • 2020
  • Lactic acid bacteria prevent the contamination of food products by inhibiting proliferation of pathogenic bacteria. This is done mainly by the production of lactic acid and antimicrobial peptides (AMPS) known as bacteriocins. The interest in these molecules resides in both their antimicrobial spectrum and safety for human health. The application of bacteriocins or producer strains has been considered to avoid the development of pathogenic bacteria, as most bacteriocins have significant inhibitory activity against food pathogens, such as Listeria monocytogenes. This article describes the classification, structure, mode of action, biosynthesis, and main applications of bacteriocins in different fields: agri-food, aquaculture, and medicine.

Diverse Antibacterial Activity of Pectobacterium carotovorum subsp. carotovorum Isolated in Korea

  • Roh, Eun-Jung;Lee, Seung-Don;Lee, Yong-Hoon;Ra, Dong-Su;Choi, Jae-Hyuk;Moon, Eun-Pyo;Heu, Sung-Gi
    • Journal of Microbiology and Biotechnology
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    • v.19 no.1
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    • pp.42-50
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    • 2009
  • Fifty-four Pectobacterium carotovorum subsp. carotovorum strains isolated in Korea were characterized by a spectrum of antibacterial activities against 7 indicator strains chosen to represent various regions and host plants. All P. carotovorum subsp. carotovorum isolates tested could be grouped into 4 classes depending on the pattern of antibacterial substance production. All tested strains had DNA fragment(s) homologous to the genes encoding carotovoricin and 21 of them had genes homologous to DNA invertase. Sixteen strains had genes homologous to the genes encoding carocin S1. Several isolates produced antibacterial substances active against strains in Brenneria, Pantoea, and Pectobacterium genera that belonged formerly to the genus Erwinia. Strains in Pseudomonas or Xanthomonas sp. were not sensitive to the antibacterial substances produced by P. carotovorum subsp. carotovorum, except for X. albilineans that was sensitive to antibacterial substances produced by most strains in P. carotovorum subsp. carotovorum and P. betavasculorum KACC10056. These results demonstrated the diverse patterns of antibacterial substance production and the possibility of the existence of new antibacterial substance(s) produced by P. carotovorum subsp. carotovorum isolated in Korea.

Prevention of Fungal Contamination during Cheese Ripening - Current Situation and Future Prospects (치즈 숙성 중의 곰팡이 오염 방제 - 현황과 전망)

  • Jung, Hoo Kil;Choi, Ha Nuel;Oh, Hyun Hee;Huh, Chang Ki;Yang, Hee Sun;Oh, Jeon Hui;Park, Jong Hyuk;Choi, Hee Young;Kim, Kyoung Hee;Lee, Seung Gu
    • Journal of Dairy Science and Biotechnology
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    • v.30 no.2
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    • pp.75-81
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    • 2012
  • Molds cause severe cheese deterioration, even though some white and blue molds are used for the manufacture of Camembert and Blue cheese, respectively. The species of Geotrichum, Moniliella, Aspergillus, Penicillium, Mucor, Fusarium, Phoma, and Cladosporium are the main fungi that affect contamination during cheese ripening. Once deteriorated by fungal spoilage, cheese becomes toxic and inedible. Fungal deterioration of cheese decreases the nutritional value, flavor profiles, physicochemical and organoleptic properties, and increases toxicity and infectious disease. Fungal contamination during cheese ripening is highly damaging to cheese production in Korean farmstead milk processing companies. Therefore, these companies hesitate to develop natural and ripened cheese varieties. This article discusses the recent and ongoing developments in the removal techniques of fungal contamination during cheese ripening. There are 2 categories of antifungal agents: chemical and natural. Major chemical agents are preservatives (propionic acid, sodium propionate, and calcium propionate) and ethanol. Among the natural agents, grapefruit seed extract, phytoncide, essential oils, and garlic have been investigated as natural antifungal agents. Additionally, some studies have shown that antibiotics such as natamycin and Delvocid$^{(R)}$, have antifungal activities for cheese contaminated with fungi. Microbial resources such as probiotic lactic acid bacteria, Propionibacterium, lactic acid bacteria from Kimchi, and bacteriocin are well known as antifungal agents. In addition, ozonization treatment has been reported to inhibit the growth activity of cheese-contaminating fungi.

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Characteristics of Pichia anomala K15 Producing Killer Toxin Isolated from Traditional Nuruk (전통 누룩으로부터 분리된 Killer Toxin 생산 균주 Pichia anomala K15의 특성)

  • Jung, Hee-Kyoung;Park, Chi-Duck;Lee, Gee-Dong;Park, Seung-Chun;Park, Hwan-Hee;Hong, Joo-Heon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.8
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    • pp.1077-1082
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    • 2007
  • In this study, killer yeasts were isolated from traditional Nuruk to improve storage and suppress contaminant in food industry. Among killer yeasts, yeast K15 showed strong killer toxin activity and inhibited growth of Salmonella Typhimurium and Vibrio parahaemolyticus. Killer yeast K15 was identified with Pichia anomala by the Microlog TM 4.0 identification system and homology of the ITS sequence. Killer toxin generated from P. anomala K15 was inactivated by pronase E and suggested to be a protein. Therefore killer toxin of P. anomala K15 was thought to be safe in human such as bacteriocin. P. anomala K15 was sufficient for growth in 50% glucose and could be used to prevent contaminant in initial stages of alcohol beverage fermentation.

Antimicrobial Effects of Lactic Acid Bacteria Isolated from Tibetan Yogurt against Foodborne Pathogenic Bacteria (티베트 요거트에서 분리한 유산균의 병원성 세균 항균 효과 연구)

  • Gho, Ju Young;Lee, Jiyeon;Choi, Hanhee;Park, Sun Woo;Kang, Seok-Seong
    • Journal of Dairy Science and Biotechnology
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    • v.39 no.3
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    • pp.121-127
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    • 2021
  • Yogurt is produced by bacterial fermentation of milk and contains lactic acid bacteria (LAB), which produce various metabolites such as organic acid, hydrogen peroxide, and bacteriocin. This study aimed to investigate cell-free supernatants (CFS) of LAB isolated from Tibetan yogurt. CFS (TY1, TY2, TY3, TY4, TY5, TY6, and TY7) from selected strains of LAB were co-incubated with four different foodborne pathogenic bacteria, namely E. coli O157:H7, Listeria monocytogenes, Salmonella typhimurium, and Staphylococcus aureus. Inhibition of foodborne pathogenic bacterial growth was not affected in the presence of CFS (pH 6.5). In contrast, CFS without neutralization completely inhibited the growth of the bacteria. Furthermore, when the concentration of CFS (without neutralization) was changed to 1:4 and 1:8, a difference in inhibition was observed between Gram-positive and Gram-negative bacteria. CFS more effectively inhibited the growth of Gram-negative E. coli O157:H7 and S. Typhimurium than Gram-positive L. monocytogenes and S. aureus. These results suggest that organic acids in LAB may inhibit the growth of foodborne pathogenic bacteria, particularly Gram-negative bacteria.

Functional evaluation of sourdough containing lactic acid bacteria isolated from sliced radish kimchi (깍두기로부터 분리된 유산균으로 제조한 사워도우의 기능성 평가)

  • Lim, Eun-Seo;Kim, Young-Mog;Lee, Eun-Woo
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.180-192
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    • 2017
  • The purpose of this study is to investigate the antioxidative and antimicrobial activities of sourdough fermented with the lactic acid bacteria (LAB) isolated from sliced radish kimchi. According to 16S rRNA gene sequence analysis, the isolated lactic acid bacteria were categorized as Leuconostoc dextranicum SRK03, Lactobacillus brevis SRK15, Pediococcus halophilus SRK22, Lactobacillus acidophilus SRK30, Lactobacillus plantarum SRK38, Leuconostoc citreum SRK 42, and Lactobacillus delbrueckii SRK60 with sequence similarity of 99%. After fermentation with L. dextranicum SRK03, L. acidophilus SRK30, L. plantarum SRK38 or L. delbrueckii SRK60 and Saccharomyces cerevisiae KCTC 7246 at $30^{\circ}C$ for 24 h LAB and yeast in sourdough were present at levels of $10^9$ and $10^7CFU/g$, respectively. In particular, the titratable acidity and ethanol and exopolysaccharide contents of sourdough fermented with L. dextranicum SRK03 were also significantly (P < 0.05) higher than those of sourdough fermented with L. acidophilus SRK30, L. plantarum SRK38, or L. delbrueckii SRK60. The sourdough fermented with L. dextranicum SRK03 and L. acidophilus SRK30 showed not only good DPPH radical-scavenging capacity but anti-lipid peroxidation activity. In addition, the viable counts of Bacillus cereus ATCC 11778 and Staphylococcus aureus ATCC 6538 in sourdough during storage for 5 days at $25^{\circ}C$ were significantly (P < 0.05) lower than those of pathogenic bacteria in the control group due to the organic acids and bacteriocin produced by L. acidophilus SRK30 strain.