• Title/Summary/Keyword: Bacteriocin activity

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Screening and Characterization of Pro biotic Lactic Acid Bacteria Isolated from Korean Fermented Foods

  • Lim, Sung-Mee;Im, Dong-Soon
    • Journal of Microbiology and Biotechnology
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    • v.19 no.2
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    • pp.178-186
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    • 2009
  • To examine their potential as probiotics, acid and bile tolerance, antibiotics resistance, adhesion capacity to Caco-2 and HT-29, and antibacterial activity, of LAB isolated from Korean fermented foods such. as dongchimi, kimchi, Meju, and doenjang were assayed against foodborne pathogenic bacteria. DC 55, DC 136, DC 222, KC 21, KC 24, KC 34, KC 43, KC 117, MJ 54, MJ 301, SP 33, and SP 170 strains were resistant to acid and bile conditions. In particular, DC 55, DC 136, KC 24, KC 43, and MJ 301 strains were highly resistant to higher than 20 ${\mu}g/ml$ concentrations of vancomycin, streptomycin sulfate, or amoxicillin, whereas, DC 222, KC 21, KC 34, KC 117, MJ 54, and SP 33 strains were susceptible to lower than 2 ${\mu}g/ml$ concentrations of those antibiotics. The adhesion to HT-29 and Caco-2 cells varied with the strains tested in a strain-dependent manner. The highest level of adhesion was observed with DC 55, KC 21, KC 24, and MJ 301 strains, having higher than 50% of adhesion to HT-29 or Caco-2 cells. In addition, Staphylococcus aureus was the most sensitive to KC 21, showing an inhibition of about 70%, and the antibacterial activity of KC 21 against S. aureus resulted most likely from both organic acids and bacteriocin. Based on its phenotypic characteristics and utilization of various sugars, the KC 21 strain was identified as Lactobacillus plantarum.

Bacteriocin ("Vulnificin") Typing of Vibrio vulnificus

  • Ha, Tai-You;Lee, Jeong-Ho;Lee, Hern-Ku;Whang, Hee-Sung;Park, Jae-Seung
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.3
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    • pp.225-237
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    • 2000
  • Vibrio vulnificus, a halophilic vibrio is an estuarine gram-negative bacteria that is associated with severe and frequently fatal wound infections and life-threatening septicemia. Bacteriocins are defined as antibacterial substance produced by various species of bacteria which are usually active against closely related organisms. Bacteriocins have found widespread application in epidemiological studies as specific markers of bacteria. It was proposed by Ha et al. (1990. J. Korean. Soc. Microbiol. 25: 586.) to give the bacteriocins produced by V. vulnificus the name "vulnificins". In the present study, a total of 72 strains of V. vulnificus isolated from patients and oysters were subjected to screen potential producers and indicators of vulnificin, applying ultraviolet induction method. Sensitivity of several strains of Serratia marcesans, Pseudomonas aeruginosa, Shigella flexneri, Salmonella typhi and Yersinia enterocolitica to vulnificins were also examined out. All the tested strains of V. vulnificus produced vulnificins active against indicator strains with various different inhibitory patterns. The spectrum of vulnificin activity and sensitive spectrum of indicator strains were considerably broad. Interestingly, almost all strains of S. marcescens, P. aeruginosa, Salmonella sp., Shigella sp. and Y. enterocolitica tested were sensitive to 1-7 vulnificin(s). Taken together, the present study demonstrated that all of the isolates of V. vulnificus produced vulnificins and that 8 good vulnificin producers and 10 good indicators were detected. These strains can be employed efficiently for establishing vulnificin typing scheme of V. vulnificus and for the detection of bacteriocinogeny and sensitivity in V. vulnificus. Biological role of vulnificin remains to be further elucidated.

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Identification of Bacteriocin-Producing Lactobacillus paraplantarum First Isolated from Kimchi

  • LEE JONG HOON;KIM MUIN;JEONG DO WON;KIM MIN JUNG;KIM JEONG HWAN;CHANG HAE CHOON;CHUNG DAE KYUN;KIM HAE YEONG;KIM KYOUNG HEON;LEE HYONG JOO
    • Journal of Microbiology and Biotechnology
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    • v.15 no.2
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    • pp.428-433
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    • 2005
  • A bacteriocin-producing lactic acid bacterium with inhibitory activity against the growth of Lactobacillus plantarum was isolated from kimchi, a traditional Korean fermented vegetable. For the identification of the isolate, its 16S rDNA was sequenced. As a result, the sequence showed $99\%$ homology with those from Lactobacillus paraplantarum, Lb. plantarum, and Lactobacillus pentosus. For further identification of the isolate, the sequence of its 16S/23S rDNA spacer region was determined, and the sequence matched perfectly with that of Lb. paraplantarum. SDS­PAGE fingerprinting of whole-cell proteins of the isolate was almost identical with that of Lb. paraplantarum. The isolation and identification of Lb. paraplantarum suggest that Lb. paraplantarum is one of the lactic acid bacteria involved in kimchi fermentation.

Optimization of Streptococcus macedonicus MBF10-2 Lysate Production in Plant-based Medium by Using Response Surface Methodology

  • Andyanti, Dini;Dani, Fatin M.;Mangunwardoyo, Wibowo;Sahlan, Muhamad;Malik, Amarila
    • Microbiology and Biotechnology Letters
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    • v.47 no.2
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    • pp.220-233
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    • 2019
  • Bacterial lysates have become a common ingredient for natural health care. Lactic acid bacteria (LAB) could serve as potential candidates for lysate production: the lactic acids produced by LAB have been utilized for their moisturizing, antimicrobial, and rejuvenating effects, while other substances provide topical benefits and health effects for the skin. Our study aimed to obtain lysate from a LAB S. macedonicus MBF 10-2 through an optimized fermentation using the Response Surface Methodology. Strain MBF10-2 was cultivated in a 2L fermenter tank in de Man Rogosa and Sharpe (MRS) medium and in plant-based peptone modified MRS, i.e. Soy-peptone and Vegitone. The duration and the medium composition (dextrose and soy peptone or proteose peptone) were adjusted to obtain an optimum production of cell lysate. Central Composite Design was employed for Design Expert 7.0.0 by adjusting 3 factors: dextrose (1%, 1.5%, 2%, 2.5%, 3%), soy or proteose peptone (0.5%, 0.75%, 1%, 1.25% and 1.5%), and duration of fermentation (8, 10, 12 14, and 16 h for MRS-Soy peptone and 15, 17, 19, 21, and 23 h for MRS Vegitone). Bacteriocin-Like Inhibitor Substance activity of lysate and pH were used as indicators. The optimum condition for lysate production using MRS Soy Peptone and Vegitone are as follows: dextrose concentration 2.5%, plant-based peptone 1.25%, while optimum fermentation duration were 11.18 h (MRS Soy Peptone) and 17 h (MRS Vegitone) with a starter concentration of 10% at $OD_{600nm}$ $0.2{\pm}0.05$. However, the standard MRS medium produced better quality lysate compared to MRS plant-based peptones.

Complete genome sequence of Bacillus halotolerans F41-3 isolated from wild flower in Korea (야생화로부터 분리한 Bacillus halotolerans F41-3 균주의 전체 게놈서열)

  • Heo, Jun;Kim, Soo-Jin;Kim, Jeong-Seon;Hong, Seung-Beom;Kwon, Soon-Wo
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.306-308
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    • 2019
  • A number of Bacillus strains are known to have antimicrobial activity useful in various fields. In order to prevent Propionibacterium acnes, which is one of the factors of acne, we selected Bacillus halotolerans F41-3 which have high antimicrobial activities against P. acnes. We conducted complete genome sequencing of B. halotolerans F41-3 and analyzed genomic characteristics. This genome size is 4,144,458 bp with a G + C content of 43.76%, 4,145 total genes and 3,686 protein coding genes. Among the genes, we found that gene clulster of subtilosin, a kind of bacteriocin, synthesis and gene cluster of nickel transportation. Both of them may influence inhibition of P. acnes.

Influence of soymilk and skim milk on growth and antibacterial activity of lactic acid bacteria (유산균의 증식과 항균 활성에 관한 탈지유 및 두유의 영향)

  • Lim, Eun-Seo
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.258-267
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    • 2019
  • The purpose of this study was to investigate the effect of lactic acid bacteria (LAB) on the growth and biogenic amines (BA) formation of Enterobacter aerogenes CIH05 in skim milk and soymilk. Lactobacillus acidophilus GK20, Lactobacillus paracasei GK74, and Lactobacillus plantarum GK81 isolated from mustard kimchi did not produce BA in the decarboxylation broth. L. paracasei GK74 exhibited the highest cell viability and antimicrobial compounds producing ability in fermented skim milk and soymilk samples, while the lowest producer was L. plantarum GK81. The production yield of lactic acid, hydrogen peroxide, and bacteriocin was dependent on the species of Lactobacillus and the type of culture medium. As LAB the number of viable cells of E. aerogenes CIH05 were higher in skim milk than in soymilk. When mixed culture with L. acidophilus GK20 and L. paracasei GK74 and treated with bacteriocin solution (300 AU/ml) obtained from these strains in milk media, the cell growth and cadaverine and histamine contents of E. aerogenes CIH05 were significantly (P < 0.05) lower than the respective values in control sample.

Effects of Fermentation to Improve Hygienic Quality of Powdered Raw Grains and Vegetables Raw Grains and Vegetables Using Lactobacillus sp. Isolated from Kimchi (김치분리균주 Lactobacillus sp.를 Starter로 한 발효생식 제조에서의 위생미생물 살균효과)

  • 김동호;송현파;변명우;차보숙;신명곤
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.5
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    • pp.765-769
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    • 2002
  • Improvement of hygienic quality of powdered raw grains and vegetables by fermentation was investigated. Luc-tobacillus sp. isolated from kimchi was used as a starter. The cell counts of coliform group and SS enteric bacteria on the SS agar plate in raw grains and vegetables were 2.3$\times$103 cfu/mL and 8.6$\times$10$^3$ cfu/mL, respectively. The starlet, Lactobacillu sp., reached 10$^{7}$ cfu/mL after 48 hr in fermentation. At that time, the coliform group and enteric bacteria on the SS agar plate were gradually inactivated and eliminated after 60 hr of fermentation. During the fermentation process, pH of the suspension was lowered and acidity increased. Antimicrobial activity of the acidic supernatant of fermented raw grains and vegetal]les against the E. coli sp. and Salmonella sp. was higher than that of lactic acid solution or neutralized supernatant. Therefore, it was considered that antimicrobial effect of the fermented raw grains and vegetal]les might be accelerated by tile synergic effect of acid and bacteriocin, and liquid fermentation of powdered raw grains and vegetables will be effective for inactivation of hygienic microorganisms.

Isolation and Identification of Bacteriocin-Producing Lactic Acid Bacteria (유용 박테리오신을 생산하는 유산균의 분리와 동정)

  • Hong, Sung Wook;Bae, Hyo Ju;Chang, Jin Hee;Kim, So-Young;Choi, Eun-Young;Park, Beom Young;Chung, Kun Sub;Oh, Mi-Hwa
    • Journal of Dairy Science and Biotechnology
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    • v.31 no.2
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    • pp.153-159
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    • 2013
  • Lactic acid bacteria are microorganisms that are closely associated with human and/or animal environments, and are categorized as generally recognized as safe (GRAS) organisms due to their ubiquitous appearance in foods and their contribution to the healthy microflora of mucosal surfaces. This study was performed to isolate and identify lactic acid bacteria with antagonistic effects against food-borne pathogens. A total of 3,000 acid-producing bacteria were isolated from infant feces, cattle feces, goat feces, dog feces, pig feces, vaginal tracts, vegetables, fruits, Kimchi, Jeotgal, fermented sausages, raw milk, cheese, yogurt, Cheonggukjang, Meju, and Makgeolli cultured on MRS agar with 0.05% bromocresol purple. For the isolation of bacteriocin-producing bacteria, the diameter of the clear zone was measured on MRS agar plates. Twenty-six isolates exhibited strong antibacterial activity against indicator strains such as Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella enterica serovar Enteritidis. Lactic acid bacteria were identified as Enterococcus faecalis, Enterococcus faecium, Enterococcus hirae, Lactobacillus acidophilus, Lactobacillus amylovorus, Lactobacillus curvatus, Lactobacillus plantarum, and Pediococcus acidilactici by 16S rDNA gene sequence analysis. The results of this study suggest that the isolates could be used as potential probiotic starters for functional food applications.

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Isolation, In vitro Antibacterial Activity, Bacterial Sensitivity and Plasmid Profile of Lactobacilli

  • Lonkar, P.;Harne, S.D.;Kalorey, D.R.;Kurkure, N.V.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.9
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    • pp.1336-1342
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    • 2005
  • The present research work was conducted to evaluate the beneficial effects as well as the safety aspects of lactobacilli as probiotic. Lactobacilli were isolated from poultry faecal samples, feed samples and from some known preparations procured from poultry feed manufacturers. L. acidophilus and L. sporogenes were tested for the antibacterial activity against four poultry pathogens viz. Escherichia coli, Salmonella spp., Proteus spp. and Pseudomonas aeruginosa. Cell free supernatant (CFS) of L. acidophilus exhibited significantly higher antibacterial activity against Salmonella spp. at original pH (4.50${\pm}$0.02). At the adjusted pH (6.50${\pm}$0.02) significantly higher antibacterial activity was recorded against indicator organism except for P. aeruginosa. Likewise, L. sporogenes exhibited similar antibacterial activity at original as well as adjusted pH except for E. coli. Antibacterial activity against E. coli was significantly higher at adjusted pH than at original pH of CFS. The competitive exclusion of E. coli by lactobacilli over the intestinal epithelial cells (IEC) was checked. L. acidophilus strain I, which was of poultry origin, exhibited maximum attachment over IEC as compared to other three strains of non-poultry origin viz. L. acidophilus strain II, L. sporogenes strain I and II. Overall, L. acidophilus exhibited higher competitive exclusion as compared to L. sporogenes. All the lactobacilli of poultry origin were most sensitive to penicillin G, amoxycillin, ampicillin and chloramphenicol, least sensitive to sulphamethizole, ciprofloxacin, neomycin, norfloxacin and pefloxacin and resistant to metronidazole and nalidixic acid. The isolates from probiotic preparations were most sensitive to ampicillin, amoxycillin and tetracycline, least sensitive to sulphamethizole, norfloxacin, neomycin and ceftriazone and resistant to nalidixic acid and metronidazole. Eight of the multiple drug resistant lactobacilli isolates were studied for the presence of plasmids. Plasmids could be extracted from six isolates of lactobacilli. These plasmids could be responsible for bacteriocin production or for antibiotic resistance of the strains. The lactobacilli need further studies regarding their safety for use in the probiotic preparations.

Identification and Antimicrobial Activity Detection of Lactic Acid Bacteria Isolated from Corn Stover Silage

  • Li, Dongxia;Ni, Kuikui;Pang, Huili;Wang, Yanping;Cai, Yimin;Jin, Qingsheng
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.5
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    • pp.620-631
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    • 2015
  • A total of 59 lactic acid bacteria (LAB) strains were isolated from corn stover silage. According to phenotypic and chemotaxonomic characteristics, 16S ribosomal DNA (rDNA) sequences and recA gene polymerase chain reaction amplification, these LAB isolates were identified as five species: Lactobacillus (L.) plantarum subsp. plantarum, Pediococcus pentosaceus, Enterococcus mundtii, Weissella cibaria and Leuconostoc pseudomesenteroides, respectively. Those strains were also screened for antimicrobial activity using a dual-culture agar plate assay. Based on excluding the effects of organic acids and hydrogen peroxide, two L. plantarum subsp. plantarum strains ZZU 203 and 204, which strongly inhibited Salmonella enterica ATCC $43971^T$, Micrococcus luteus ATCC $4698^T$ and Escherichia coli ATCC $11775^T$ were selected for further research on sensitivity of the antimicrobial substance to heat, pH and protease. Cell-free culture supernatants of the two strains exhibited strong heat stability (60 min at $100^{\circ}C$), but the antimicrobial activity was eliminated after treatment at $121^{\circ}C$ for 15 min. The antimicrobial substance remained active under acidic condition (pH 2.0 to 6.0), but became inactive under neutral and alkaline condition (pH 7.0 to 9.0). In addition, the antimicrobial activities of these two strains decreased remarkably after digestion by protease K. These results preliminarily suggest that the desirable antimicrobial activity of strains ZZU 203 and 204 is the result of the production of a bacteriocin-like substance, and these two strains with antimicrobial activity could be used as silage additives to inhibit proliferation of unwanted microorganism during ensiling and preserve nutrients of silage. The nature of the antimicrobial substances is being investigated in our laboratory.