• Journal of Microbiology and Biotechnology
• /
• 제8권6호
• /
• pp.588-594
• /
• 1998

Lactic acid bacteria were isolated from Kimchi and screened for bacteriocin production. Strain SE1, identified as Lactobacillus curvatus sp., showed the strongest inhibitory activity against Lactobacillus delbrueckii subsp. delbrueckii. The bacteriocin was inactivated by amyloglucosidase, trypsin, or protease K treatment. However, it maintained its activity under heat treatment at $100^{\circ}C$ for 60 min. The production of the bacteriocin had a growth-related mode and decreased around the early-stationary phase. The optimum temperature for the growth of L. curvatus SE1 was $37^{\circ}C$; however, the optimum temperature for bacteriocin production was $30^{\circ}C$. The bacteriocin activity was decreased by treatment with methanol, butanol, acetone, or chloroform, however, it was not affected by treatment with ethanol, iso-propanol, or cyclohexane. The inhibitory activity of bacteriocin was stable over a wide range of pHs (2 to 11). The bacteriocin from L. curvatus SE1 killed the indicator strain by a bactericidal mode of action. The bacteriocin from L. curvatus SE1 was partially purified by ethanol precipitation and ion exchange chromatography. SDS-polyacrylamide gel electrophoresis was used to determine the molecular weight of the bacteriocin by the bacteriocin activity test. The apparent molecular mass of the bacteriocin produced by L. curvatus SE1 was about 14 kDa.

• 한국축산식품학회지
• /
• 제38권5호
• /
• pp.1064-1079
• /
• 2018

In this study, the antimicrobial activity of a bacteriocin produced by Enterococcus faecalis KT11, isolated from traditional Kargı Tulum cheese, was determined, and bacteriocin KT11 was partially characterized. The results showed that bacteriocin KT11 was antagonistically effective against various Gram-positive and Gram-negative test bacteria, including vancomycin- and/or methicillin-resistant bacteria. The activity of bacteriocin KT11 was completely abolished after treatment with proteolytic enzymes (proteinase K, ${\alpha}$-chymotrypsin, protease and trypsin), which demonstrates the proteinaceous nature of this bacteriocin. Additionally, bacteriocin KT11 remained stable at pH values ranging from 2 to 11 and after autoclaving at $121^{\circ}C$ for 30 min. In addition, the activity of bacteriocin KT11 was stable after treatment with several surfactants (EDTA, SDS, Triton X-100, Tween 80 and urea) and organic solvents (chloroform, propanol, methanol, ethyl alcohol, acetone, hexane and ethyl ether). Cell-free supernatant of E. faecalis KT11 was subjected to ammonium sulfate precipitation and then desalted by using a 3.5-kDa cut-off dialysis membrane. The bacteriocin activity was determined to be 711 AU/mL in the dialysate. After tricine-SDS-PAGE analysis, one peptide band, which had a molecular weight of ~3.5 kDa, exhibited antimicrobial activity. Because the bacteriocin KT11, isolated from E. faecalis KT11, exhibits a broad antimicrobial spectrum, heat stability and stability over a wide pH range, this bacteriocin can be used as a potential bio-preservative in foods. Additionally, bacteriocin KT11 alone or in combination with conventional antibiotics may provide a therapeutic option for the treatment of multidrug-resistant clinical pathogens after further in vivo studies.

• 한국축산식품학회지
• /
• 제35권4호
• /
• pp.473-478
• /
• 2015

The aim of this study was to screen and In vitro characterize the properties of bacteriocin produced by lactic acid bacteria isolated from Vietnamese fermented pork (Nem chua). One hundred and fifty LAB were isolated from ten samples of Nem chua and screened for bacteriocin-producing lactic acid bacteria. Antimicrobial activity of bacteriocin was carried out by spot on lawn method against both gram positive and gram negative bacteria. One isolate, assigned as KL-1, produced bacteriocin and showed inhibitory activity against Lactobacillus sakei, Leuconostoc mesenteroides and Enterococcus faecalis. To characterize the bacteriocin-producing strain, optimum temperature, incubation period for maximum bacteriocin production and identification of bacteriocin-producing strain were determined. It was found that the optimum cultivation temperature of the strain to produce the maximum bacteriocin activity (12,800 AU/mL) was obtained at 30℃. Meanwhile, bacteriocin production at 6,400 AU/mL was found when culturing the strain at 37℃ and 42℃. The isolate KL-1 was identified as L. plantarum. Antimicrobial activity of cell-free supernatant was completely inhibited by proteolytic enzyme of trypsin, alpha-chymotrypsin and proteinase K. Bacteriocin activity was stable at high temperature up to 100℃ for 10 min and at 4℃ storage for 2 d. However, the longer heating at 100℃ and 4℃ storage, its activity was reduced.

• 한국미생물·생명공학회지
• /
• 제22권1호
• /
• pp.7-12
• /
• 1994

One bacterial strain, that had made the largest inhibition zone at the antagonism assay and also that lost the inhibition activity by the protease treatment, was isolated from raw milk. That strain was identified as Streptococcus salivarius subsp. thermophilus. The specific growht rate of this strain was maximum at 45$\circ $C. However, at this temperature the strain produced no bacteriocin. The bacteriocin activity was quite stable even at high temperature. Moreover, the activity of the vacteriocin was sensitive to proteases. but not to $\alpha $-amylase, DNase I, or RNase.

• 한국미생물·생명공학회지
• /
• 제17권2호
• /
• pp.94-99
• /
• 1989

1. L. acidophilus 균주에 작용하는 bacteriocin을 생산하는 Lactobacillus속 균 7주를 발견하였다. bacteriocin 생산균주는 1주가 L. gasseri였고, 그 외에는 모두 L. acidophilus였다. 2. 조사한 bacteriocin생산 6균주 중 5균주는 대수발육 중기 혹은 후기 내지 정상기의 초기에 최대의 bacteriocin 활성을 나타내었으나, 그 후 급격히 활성이 감소하였다. 3. 이들의 bacteriocin은 단백분해효소 처리로 용이하게 실활하는 단백질성임을 나타내었으나, 핵산분해 효소에는 저항성이었다. 또 이들의 bacteriocin은 지금 까지 보고되고 있는 Lactobacillus속의 bacteriocin과 다르며, 정도의 차는 있으나 전부 10$0^{\circ}C$ 가열에 감수성이 있었다. 4. L. acidophilus ATCC 9857 및 L. acidophilus ATCC 4357이 생산하는 bacteriocin은 diethylether 처리에 활성이 감소하므로 지질을 함유함을 시사하였다. 5. L. acidophilus A4가 생산하는 bacteriocin은 지시균의 세포벽 외층의 규칙적 배열구조에 흡착함을 시사하였다. 또 L. acidophilus ATCC 4355가 생산하는 bacteriocin은 지시균의 세포벽에는 흡착하지 않음을 알 수 있었다.

• 한국식품위생안전성학회지
• /
• 제10권2호
• /
• pp.65-71
• /
• 1995

Bacteriocins from lactic acid bacteria have attracted much attention in recent years because of their useful worth in increasing safety and extending shelf life of foods. These substances show an inhibitory effect against some food spoilage bacteria and food-borne pathogens. The inhibitory effect fo the bacteriocin produces by lactic acid bacteria against Listeria monocytogenes(L. monocytogenes) was examined in this study. The culture supernatants of 5 kinds of bacteria among the 10 kinds of testes lactic acid bacteria had the inhibitory activity against Listeria sp., various Gram positive and Gram negative bacteria. Bacteriocin produced by Lactobacillus plantarum(Lact. plantarum) LMG 7945 was the most active toward L. monocytogenes. Bacteriocin production of the Lact. plantarum LMG 7945 cultured on MRS broth was increased late logarithmic phase over early stationary phase. This bacteriocin was stable at heat treatment and acidic pH relatively; The activity was retained after heating at 121$^{\circ}C$ for 15min and was active in the pH range of 2~4 but was lost above pH 5.

• Journal of Microbiology and Biotechnology
• /
• 제7권6호
• /
• pp.409-416
• /
• 1997

Leuconostoc sp. LAB145-3A isolated from kimchi produced a bacteriocin which was active against food pathogens, such as Listeria monocytogenes, Enterococcus faecalis, and E. faecium. Bacteriocin production occurred during the early exponential phase of growth and was stable upto the late stationary phase of growth. Optimum conditions for bacteriocin production were $37^{\circ}C$ with an initial pH of 7.0. The bacteriocin of LAB145-3A was sensitive to proteases, but stable for solvents, pH change and heat treatment. It was stable even at autoclaving temperature for 15 min. The bacteriocin exhibited a bactericidal mode of action against Lactobacillus curvatus LAB170-12. The bacteriocin produced by Leuconostoc sp. LAB145-3A was purified by CM-cellulose cation exchange column chromatography and Sephadex G-50 gel filtration. The purification resulted in an approximate 10,000-fold increase in the specific activity. Approximately 4% of the initial activity was recovered. Purified bacteriocin exhibited a single band on the SDS-PAGE with an apparent molecular weight of 4,400 daltons. This bacteriocin was named leucocin K. Leuconostoc sp. LAB145-3A had two residential plasmids with molecular sizes of 23 kb and 48 kb. A comparison of plasmid profiles between LAB145-3A and its mutants revealed that the 23 kb plasmid (pCA23) was responsible for bacteriocin production and immunity to the bacteriocin in Leuconostoc sp. LAB145-3A.

• 한국축산식품학회지
• /
• 제41권1호
• /
• pp.164-171
• /
• 2021

Listeria monocytogenes is a representative foodborne pathogen and causes listeriosis. Enterococcus faecium CJNU 2524 was confirmed to produce a bacteriocin with anti-listerial activity. To establish optimal culture conditions for the production of the bacteriocin from E. faecium CJNU 2524, different media (MRS and BHI broth) and temperatures (25℃, 30℃, and 37℃) were investigated. The results showed that the optimal culture conditions were MRS broth and 25℃ or 30℃ temperatures. The crude bacteriocin was stable in a broad range of pH conditions (2.0-10.0), temperatures (60℃-100℃), and organic solvents (methanol, ethanol, acetone, acetonitrile, and chloroform). The bacteriocin activity was abolished when treated with protease but not α-amylase or lipase, indicating the proteinaceous nature of the bacteriocin. Finally, the bacteriocin showed a bactericidal mode of action against L. monocytogenes. Therefore, it can be a biopreservative candidate for controlling L. monocytogenes in dairy and meat products.

• 한국미생물·생명공학회지
• /
• 제26권6호
• /
• pp.523-528
• /
• 1998

본 연구에서는 토마토에서 bacteriocin 생산균주를 분리하여 균주동정 및 bacteriocin 정제를 행하였다. 분리균주는 Gram 염색, catalase test, 포자생성유무, API 20 STREP. kit 등을 통하여 Enterococus faecium으로 잠정 동정하였다. 분리균주가 생산하는 항균성물질은 Gram 양성 세균인 Listeria manocytogmes, Leuonostoc mesenteroides, Lactobacillus plantarum, Streptococus agalartiae, Streptococus pyrogenes, Gram 음성 세균인 Pseudomonas aeruginosa등에 대해서 항균력을 나타내었다. 단백질 분해 효소인 proteinase K에 의해서 그 활성이 소실되고, catalase에 의해서는 항균력이 유지되므로 단백질성 bacteriocin임을 확인할 수 있었다. Bacteriocin 정제를 위하여 ethanol침전, CM Sepharose CL-6B 양이온교환 chromatography, Sephacryl S-100 HR gel filtration의 순으로 정제하여 초기 배양액의 bacteriocin활성 대비 35.8배의 정제도를 얻었다. 13% SDS-polyacrylamide gel전기 영동을 통하여 단일 band를 확인할 수 있었으며 분자량은 약 51 kDa 정도_로 추정되었다. Crude bacteriocin의 열 안정성은 10$0^{\circ}C$에서 1시간 처리하였을 때의 잔존활성이 3.3%로 비교적 낮았으며, pH에 대해서는 pH2~7에서 비교적 안정하였다. Bacteriocin의 항균작용을 알아보기 위하여 피검균인 Leu. mesenteroides를 초기농도 1.0$\times$$10^{9}$ CFU/$m\ell$로 접종한 배지에 bacteriocin을 첨가하여 배양하였을 때 4시간 이후에 완전히 사멸되므로 bacteriocidal 작용을 하는 것으로 추정되었다.

• Journal of Microbiology and Biotechnology
• /
• 제14권3호
• /
• pp.503-508
• /
• 2004

The antimicrobial substance produced by Lactobacillus bulgaricus was inactivated by protease. It showed inhibitory activity against Staphylococcus aureus ATCC6538, Streptococcus agalactiae ATCC14364, some Gram-positive and Gram-negative bacteria, and characteristics of a bacteriocin. The optimal temperature and culture time for the production of bacteriocin were $30^{\circ}C$ and 10 h, respectively, in the culture of L. bulgaricus. The bacteriocin production started in the exponential phase and reached a maximum at the early stationary phase. Using Staph. aureus ATCC6538 and Strep. agalactiae ATCC14364, known as common bovine mastitis pathogens, as indicator strains for determination of the bacteriocin activity, the antimicrobial activity of the bacteriocin was found to be stable in acidic and neutral pH's (2- 7) even at lOOT, whereas it was lost at high pH (10- 11) and $100^{\circ}C$. The mode of action for the antimicrobial activity was bacteriocidal, and the molecular weight determined by SDS-PAGE and overlay method was 14 kDa.