• Title/Summary/Keyword: Lactobacillus identification

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Biochemical and Molecular Identification of Antibacterial Lactic Acid Bacteria Isolated from Kimchi (김치에서 항균활성 유산균의 분리 및 동정)

  • Kim, Soo-Young;Kim, Jong-Doo;Son, Ji-Soo;Lee, Si-Kyung;Park, Kab-Joo;Park, Myeong-Soo
    • Korean Journal of Food Science and Technology
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    • v.43 no.4
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    • pp.446-452
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    • 2011
  • Total 480 lactic acid-producing bacteria were isolated from five kinds of kimchi, and their antibacterial activity was tested against Salmonella enterica serovar Typhimurium, Bacillus subtilis, and Pseudomonas aeruginosa using an agar diffusion assay. Among them, 340 isolates showed a halo on MRS agar against one or more indicator strains, which were identified using multiplex PCR, an API 50CHL kit, and a 16S rDNA sequence analysis. As a result, 169 Lactobacillus plantarum, 20 Lactobacillus fermentum, two Lactobacillus paracasei ssp. paracasei, two Lactobacillus sp., and 15 Pediococcus sp. were identified. This may be the first report on the isolation of antibacterial Lactobacillus fermentum from kimchi.

Identification of Bacteriocin-producing Lactic Acid Bacteria from Kimchi and Partial Characterization of their Bacteriocin

  • Ha, Duk-Mo;Cha, Dong-Soo
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.305-315
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    • 1994
  • Nineteen strains of bacteriocin-producing lactic acid bacteria were isolated from 432 Kimchi samples, and identified by the comprehensive biochemical and morphological tests verifying their cellular fatty acid composition. Using partially purified bacteriocins from these isolates, their inhibitory activities against other lactic acid bacteria and some pathogens, and sensitivity to enzyme and heat treatments were tested. The isolates were identified as Lactobacillus plantarum (2 strains), L curvatus (2 starins), L brevis (2 strains), Enterococcus faecium (6 strains), Leuconostoc mesenteroides subsp. mesenteroides (1 strain) and Lactobacillus sp. (6 strains). The bacteriocins produced by E. faecium strains provided the broadest spectrum of inhibition, affecting against other Gram-positive bacteria including lactic acid bacteria and health-threatening bacteria such as Clostridium perfringens and Listeria monocytogenes. The bacteriocins of Lactobacillus sp., L plantarum and L brevis strains were capable of inhibiting many strains of the lactic acid bacteria, whereas those of L curvatus and L mesenteroides subsp. mesenteroides strains were only inhibitory to a few strains. Generally, the inhibitory activities of both E. faecium and Lactobacillus sp. strains were greater than those of other producer strains. The bacteriocins from the isolates were sensitive to several proteolytic enzymes, and those of L curvatus and L mesenteroides subsp. mesenteroides were also sensitive to lipase and $\alpha$-amylase as well as to proteolytic enzymes. The bacteriocins from the strains of Lactobacillus sp. and a strain of L. brevis were resistant to autoclaving.

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Molecular Identification of Vaginal Lactobacillus spp. Isolated from Korean Women

  • CHANG, CHUNG EUN;SYLVIA I. PAVLOVA;LIN TAO;EUN-KI KIM;SEUNG CHUL KIM;HYUN SHIK YUN;JAE-SEONG SO
    • Journal of Microbiology and Biotechnology
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    • v.12 no.2
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    • pp.312-317
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    • 2002
  • Indigenous lactobacilli were isolated from vaginas of Korean women for possible use in ecological treatment of bacterial vaginosis. Vaginal swab samples were obtained from a gynecological clinic and streaked on Rogosa SL agar plates to select the most predominant lactobacilli in each sample. The preliminary identification of the isolates as lactobacilli was based on microscopic observation of Gram-positive rod-shaped cell morphology. The initial characterization was performed on 108 isolates in terms of their cell surface hydrophobicity (CSH), antimicrobial activity, and hydrogen peroxide (H₂O₂) production capability, and 10 isolates were then selected for further molecular identification. For a rapid procedure to identify lactobacilli, polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses of the l6S rRNA genes were applied. The 10 selected lactobacilli and 9 different reference strains of Lactobacillus spp. were characterized by PCR-RFLP where the amplified l6S rDNA was digested with 7 different restriction endonucleases prior to analysis. DNA sequencing of the 16S rRNA gene of one particular isolate, KLB 46, that had been identified as L. crispatus by the PCR-RFLP analysis, further confirmed its identity as L. crispatus.

Identification and Monitoring of Lactobacillus delbrueckii Subspecies Using Pangenomic-Based Novel Genetic Markers

  • Kim, Eiseul;Cho, Eun-Ji;Yang, Seung-Min;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.31 no.2
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    • pp.280-289
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    • 2021
  • Genetic markers currently used for the discrimination of Lactobacillus delbrueckii subspecies have low efficiency for identification at subspecies level. Therefore, our objective in this study was to select novel genetic markers for accurate identification and discrimination of six L. delbrueckii subspecies based on pangenome analysis. We evaluated L. delbrueckii genomes to avoid making incorrect conclusions in the process of selecting genetic markers due to mislabeled genomes. Genome analysis showed that two genomes of L. delbrueckii subspecies deposited at NCBI were misidentified. Based on these results, subspecies-specific genetic markers were selected by comparing the core and pangenomes. Genetic markers were confirmed to be specific for 59,196,562 genome sequences via in silico analysis. They were found in all strains of the same subspecies, but not in other subspecies or bacterial strains. These genetic markers also could be used to accurately identify genomes at the subspecies level for genomes known at the species level. A real-time PCR method for detecting three main subspecies (L. delbrueckii subsp. delbrueckii, lactis, and bulgaricus) was developed to cost-effectively identify them using genetic markers. Results showed 100% specificity for each subspecies. These genetic markers could differentiate each subspecies from 44 other lactic acid bacteria. This real-time PCR method was then applied to monitor 26 probiotics and dairy products. It was also used to identify 64 unknown strains isolated from raw milk samples and dairy products. Results confirmed that unknown isolates and subspecies contained in the product could be accurately identified using this real-time PCR method.

Identification of Lactic Acid Bacteria and Changes of Organic Acid during Aging of Traditional Kyungsando Squid sikhe (경상도 전통 마른 오징어 식해로부터 젖산균의 분리, 동정 및 숙성과정 중 유기산의 변화)

  • Lee, Hee-Duck;Choi, Hee-Jin;Kim, Sung;Seong, Tae-Su;Choi, Cheong
    • Applied Biological Chemistry
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    • v.44 no.3
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    • pp.167-172
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    • 2001
  • The objective of this study is to investigate identification of lactic acid bacteria and changes of organic acid during aging of traditional Kyungsando Squid sikhe. Lactobacillus brevis SH-1, Lactobacillus plantarum SH-2, and Leuconostoc mensenteroides SH-3, which were isolated from Kyungsando Squid sikhe were selected for fermentation. The viable cell and lactic acid bacteria counts of squid sikhe was increased on fermentation days, and slowly decreased after 10 days of fermentation. The viable cell and lactic acid bacteria counts of standard sikhe at 6 days fermentation was $3.7{\times}10^9$ and $8.2{\times}10^8\;CFU/g$, respectively. The organic acid of squid sikhe consisted of 7 kinds including lactic acid during fermentation. The contents of citric acid, succinic acid, and lactic acid gradually increased during fermentation, and especially, increasing of lactic acid remarkably increased than the different organic acid. The contents of acetic acid and tartaric acid slowly increased after 7 days of fermentation.

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Isolation and identification of Lactobacillus inhibiting the production of halitosis by anaerobic bacteria (구취를 유발하는 혐기성 세균의 증식을 억제하는 유산 간균의 분리 및 동정)

  • Kim, Mi-Hyung;Kim, Seon-Mi
    • Journal of Korean society of Dental Hygiene
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    • v.4 no.2
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    • pp.153-163
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    • 2004
  • There are normal inhabitants doing medically useful functions in the body. There are many kinds of bacteria performing specific functions in the oral cavity. Two strains of lactic acid bacteria were isolated from normal inhabitants of children 's oral cavity, which inhibited the the production of halitosis by anaerobic bacteria. The authors identified the isolates by the lest using API 50 CHL medium kit. 1. Two isolates were Gram-positive bacilli and produced hydrogen peroxide. 2. The optical density was 1.286 in the supernatant of Fusobacterium nucleatum after vortexing for 30 minutes, whereas in the supernatant of combined Fusobacterium nucleatum and each isolate, they were reduced to 0.628 and 0.497, which the percentages of coaggregation between them were 29.4% and 57.8%, respectively. 3. The optical density of Fusobacterium nucleatum precipitate was 1.794 in the culture media containing cysteine and $FeSO_4$, being reduced to 1.144 and 0.915 in the coaggregated precipitates of Fusobacterium nucleatum and each isolate. 4. The optical density of Porphyromonas gingivalis precipitate was 1.932 in the culture media, being reduced to 1.170 and 1.266 in the coaggregated precipitates of Porphyromonas gingivalis and each isolate. 5. When two isolates were tested with API 50 CHL medium kit, those were identified as Lactobaciallius salivarius and Lactobacillus delbrueckii subsp. lactis.

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Changes in Lactic Acid Bacteria of Squid with Low Salt during Fermentation (저염 오징어젓갈 숙성중 젖산균의 변화)

  • Jo, Jin-Ho;Oh, Se-Wook;Kim, Young-Myoung;Chung, Dong-Hyo;Kim, Joung-Im
    • Korean Journal of Food Science and Technology
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    • v.29 no.6
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    • pp.1208-1212
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    • 1997
  • Isolation and identification of predominant microorganisms in fermented squid with low salt were carried out during fermentation at $10^{\circ}C$. Dominant strains were lactic acid bacteria(80%) including Lactobacillus plantarum, Lactobacillus brevis, Leuconostoc sp., Pediococcus sp. and Streptococcus sp. Leuconostoc, Pediococcus and Streptococcus were shown in the early stage of fermentation and gradually increased until optimum stage of squid fermentation, and then decreased rapidly. Lactobacillus grew lastingly during fermentation. Yeasts were detected in the middle stage of fermentation and shown rapid increase after the last stage of fermentation, suggesting that yeasts participate in putrefaction of fermented squid with low salt.

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Isolation and Identification of Lactic Acid Bacteria from Spent Mushroom Substrate for Silage Making and Determination of Optimal Medium Conditions for Growth

  • Kim, Young-Il;Kwak, Wan-Sup
    • Journal of Animal Science and Technology
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    • v.54 no.6
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    • pp.435-442
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    • 2012
  • This study was conducted to isolate and identify the lactic acid bacteria (LAB) from spent mushroom substrates (SMS) for the effective anaerobic fermentation to utilize SMS as an animal feed and to determine the optimal medium conditions for their growth. At first, a total of 23 strains were isolated from the ensiled SMS based on the LAB counts and pH tested. Then, a total of 16 strains which rapidly produce lactate and decreased the pH, were selected for a screening test. The optical density (OD), pH, and yellow clear zone were tested for the selected 16 strains. Among the strains, KU5 strain had wider yellow clear zone and lower pH and KU13 strain had higher OD at 24 hr of incubation and wider yellow clear zone compared to other strains and control strain (Lactobacillus plantarum KCCM 12116). Accordingly, KU5 and KU13 strains were finally selected. The KU5 and KU13 were identified as Lactobacillus plantarum by the 16S rRNA sequencing. The KU5 strain was named as Lactobacillus plantarum KU5, and the KU13 strain was named as Lactobacillus plantarum KU13. Lactobacillus plantarum KU5 and Lactobacillus plantarum KU13 were registered at the National Center for Biotechnology Information (NCBI). Access number of Lactobacillus plantarum KU5 was HQ542227 and that of Lactobacillus plantarum KU13 was HQ542228. The optimal medium conditions for growth of KU5 and KU13 were soybean meal 2% and formulated feed 2%, respectively.

Rapid Identification of Lactobacillus plantarium in Kimchi Using Polymerase Chain Reaction

  • Kim, Tae-Woon;Min, Sung-Gi;Choi, Dong-Hun;Jo, Jae-Sun;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.881-884
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    • 2000
  • A polymerase chain reaction (PCR) was performed to rapidly identify Lactobacillus plantarum from type strains and kimchi samples. The PCR experiments were carried out using specific oligonucleotide primer sets based on the 16S rRNA gene sequences of L. plantarum. The expected DNA amplificate of 419 bp was obtained when either purified DNA or whole cells of L. plantarum strains reacted with LP primers, yet not with any of the other strains. The PCR product was confirmed by DNA sequencing. Accordingly, since the PCR method used is simple, specific, and rapid, it will be useful for monitoring and evaluation L. plantarum in the mixed microbial population found in kimchi.

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