• Title/Summary/Keyword: identification of lactic acid bacteria

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Identification and Characterization of Hydrogen Peroxide-generating Lactobacillus fermentum CS12-1

  • Kang, Dae-Kyung;Oh, H.K.;Ham, J.-S.;Kim, J.G.;Yoon, C.H.;Ahn, Y.T.;Kim, H.U.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.1
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    • pp.90-95
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    • 2005
  • Lactic acid bacteria were isolated from silage, which produce high level of hydrogen peroxide in cell culture supernatant. The 16S rDNA sequences of the isolate matched perfectly with that of Lactobacillus fermentum (99.9%), examined by a 16S rDNA gene sequence analysis and similarity search using the GenBank database, thus named L. fermentum CS12-1. L. fermentum CS12-1 showed resistance to low pH and bile acid. The production of hydrogen peroxide by L. fermentum CS12-1 was confirmed by catalase treatment and high-performance liquid chromatography. L. fermentum CS12-1 accumulated hydrogen peroxide in culture broth as cells grew, and the highest concentration of hydrogen peroxide reached 3.5 mM at the late stationary growth phase. The cell-free supernatant of L. fermentum CS12-1 both before and after neutralization inhibited the growth of enterotoxigenic Escherichia coli K88 that causes diarrhea in piglets.

Isolation and Identification of Lactic Acid Bacteria from Traditional Dairy Products in Baotou and Bayannur of Midwestern Inner Mongolia and q-PCR Analysis of Predominant Species

  • Wang, Dan;Liu, Wenjun;Ren, Yan;De, Liangliang;Zhang, Donglei;Yang, Yanrong;Bao, Qiuhua;Zhang, Heping;Menghe, Bilige
    • Food Science of Animal Resources
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    • v.36 no.4
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    • pp.499-507
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    • 2016
  • In this study, traditional culture method and 16S rRNA gene analysis were applied to reveal the composition and diversity of lactic acid bacteria (LAB) of fermented cow milk, huruud and urum from Baotou and Bayannur of midwestern Inner Mongolia. Also, the quantitative results of dominant LAB species in three different types of dairy products from Baotou and Bayannur were gained by quantitative polymerase chain reaction (q-PCR) technology. Two hundred and two LAB strains isolated from sixty-six samples were identified and classified into four genera, namely Enterococcus, Lactococcus, Lactobacillus, Leuconostoc, and twenty-one species and subspecies. From these isolates, Lactococcus lactis subsp. lactis (32.18%), Lactobacillus plantarum (12.38%) and Leuconosto mesenteroides (11.39%) were considered as the dominated LAB species under the condition of cultivating in MRS and M17 medium. And the q-PCR results revealed that the number of dominant species varied from samples to samples and from region to region. This study clearly shows the composition and diversity of LAB existing in fermented cow milk, huruud and urum, which could be considered as valuable resources for LAB isolation and further probiotic selection.

Biotransformation of major ginsenosides in ginsenoside model culture by lactic acid bacteria

  • Park, Seong-Eun;Na, Chang-Su;Yoo, Seon-A;Seo, Seung-Ho;Son, Hong-Seok
    • Journal of Ginseng Research
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    • v.41 no.1
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    • pp.36-42
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    • 2017
  • Background: Some differences have been reported in the biotransformation of ginsenosides, probably due to the types of materials used such as ginseng, enzymes, and microorganisms. Moreover, most microorganisms used for transforming ginsenosides do not meet food-grade standards. We investigated the statistical conversion rate of major ginsenosides in ginsenosides model culture during fermentation by lactic acid bacteria (LAB) to estimate possible pathways. Methods: Ginsenosides standard mix was used as a model culture to facilitate clear identification of the metabolic changes. Changes in eight ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, Rg1, and Rg2) during fermentation with six strains of LAB were investigated. Results: In most cases, the residual ginsenoside level decreased by 5.9-36.8% compared with the initial ginsenoside level. Ginsenosides Rb1, Rb2, Rc, and Re continuously decreased during fermentation. By contrast, Rd was maintained or slightly increased after 1 d of fermentation. Rg1 and Rg2 reached their lowest values after 1-2 d of fermentation, and then began to increase gradually. The conversion of Rd, Rg1, and Rg2 into smaller deglycosylated forms was more rapid than that of Rd from Rb1, Rb2, and Rc, as well as that of Rg1 and Rg2 from Re during the first 2 d of fermentation with LAB. Conclusion: Ginsenosides Rb1, Rb2, Rc, and Re continuously decreased, whereas ginsenosides Rd, Rg1, and Rg2 increased after 1-2 d of fermentation. This study may provide new insights into the metabolism of ginsenosides and can clarify the metabolic changes in ginsenosides biotransformed by LAB.

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.

Identification of Coccoidal Bacteria in Traditional Fermented Milk Products from Mongolia, and the Fermentation Properties of the Predominant Species, Streptococcus thermophilus

  • Ren, Yan;Liu, Wenjun;Zhang, Heping
    • Food Science of Animal Resources
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    • v.35 no.5
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    • pp.683-691
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    • 2015
  • The objective of this study was to identify the coccoidal bacteria present in 188 samples of fermented yaks’, mares’ and cows’ milk products collected from 12 different regions in Mongolia. Furthermore, we evaluated the fermentation properties of ten selected isolates of the predominant species, Streptococcus (S.) thermophiles, during the process of milk fermentation and subsequent storage of the resulting yoghurt at 4℃. Overall, 159 isolates were obtained from 188 samples using M17 agar. These isolates were presumed to be lactic acid bacteria based on their gram-positive and catalase-negative properties, and were identified to species level using 16S rRNA gene sequence analysis. These coccoid isolates were distributed in four genera and six species: Enterococcus (E.) durans, Enterococcus (E.) faecalis, Lactococcus (Lac.) subsp. lactis, Leuconostoc (Leuc.) lactis, Leuconostoc (Leuc.) mesenteroides. subsp. mesenteroides and S. thermophilus. Among these S. thermophilus was the most common species in most samples. From evaluation of the fermentation characteristics (viable counts, pH, titratable acidity [TA]) of ten selected S. thermophilus isolates we could identify four isolates (IMAU 20246, IMAU20764, IMAU20729 and IMAU20738) that were fast acid producers. IMAU20246 produced the highest concentrations of lactic acid and formic acid. These isolates have potential as starter cultures for yoghurt production.

Identification of the Cell-envelope Proteinase of Lactic Acid Bacteria Isolated from Kimchi. (김치 유래 젖산균의 Cell-envelope Proteinase 존재 확인)

  • 이유진;최재연;이형주;장해춘;김정환;정대균;김영석;김소미;이종훈
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.116-122
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    • 2002
  • The partial 165 rDNA sequences of 6 lactic acid bacterial strains isolated from Kimchi were determined. Two strains were Leuconostoc mesenteroides and the rest were incorrectly classified and turned out to be Lactobacillus. As the case of dairy lactic acid bacteria, the strains isolated from Kimchi also had cell-envelope proteinase (CEP) activity. As the result of partial CEP gene amplification with CEP-specific primers, the expected 1.2-kb amplificate was obtained not from Leu. mesenteroides but from Lactobacillus strains. The deduced amino acid sequence of PCR product amplified from the genomic DNA of Lactobacillus pentosus KFR1821 showed 95% and 92% homology with those of PrtPs from Lactococcus lactis subsp. cremoris and Lactobacillus paracasei subsp. paracasei, respectively. The PCR amplificate was used as a probe and the result of Southern hybridization illuminated the location of CEP gene in chromosomal DNA of Lb. pentosus KFR1821.

Classification of Isolates Originating from Kimchi Using Carbon-source Utilization Patterns

  • LEE, JUNG-SOOK;CHANG OUK CHUN;MIN-CHUL JUNG;WOO-SIK KIM;HONG-JOONG KIM;MARTIN HECTOR;SAM-BONG KIM;CHAN-SUN PARK
    • Journal of Microbiology and Biotechnology
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    • v.7 no.1
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    • pp.68-74
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    • 1997
  • One hundred and eighty two lactic acid bacteria, isolated mainly from kimchi, including reference strains were examined for their ability to utilize 95 carbon sources. The test strains were assigned to 5 major, 1 minor and 12 single-membered clusters based on the $S_{SM}$, UPGMA algorithm (at similarity of $80{\%}$). These aggregate clusters were equivalent to the genus Leuconostoc (aggregate cluster M and N), the genus Lactobacillus (aggregate cluster Q and R), and the genera Lactobacillus and Leuconostoc (aggregate cluster O and P) according to the database of the Biolog system. This study demonstrates that rapid identification and classification of isolates originating from kimchi can be achieved on the basis of such carbon source utilization tests.

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Identification of LAB and Fungi in Laru, a Fermentation Starter, by PCR-DGGE, SDS-PAGE, and MALDI-TOF MS

  • Ahmadsah, Lenny S.F.;Kim, Eiseul;Jung, Youn-Sik;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.1
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    • pp.32-39
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    • 2018
  • Samples of Laru (a fermentation starter) obtained from the upper part of Borneo Island were analyzed for their lactic acid bacteria (LAB) and fungal diversity using both a culture-independent method (PCR-DGGE) and culture-dependent methods (SDS-PAGE and MALDI-TOF MS). Pediococcus pentosaceus, Lactobacillus brevis, Saccharomycopsis fibuligera, Hyphopichia burtonii, and Kodamaea ohmeri were detected by all three methods. In addition, Weissella cibaria, Weissella paramesenteroides, Leuconostoc citreum, Leuconostoc mesenteroides, Lactococcus lactis, Rhizopus oryzae/Amylomyces rouxii, Mucor indicus, and Candida intermedia were detected by PCR-DGGE. In contrast, Lactobacillus fermentum, Lactobacillus plantarum, Pichia anomala, Candida parapsilosis, and Candida orthopsilosis were detected only by the culture-dependent methods. Our results indicate that the culture-independent method can be used to determine whether multiple laru samples originated from the same manufacturing region; however, using the culture-independent and the two culture-dependent approaches in combination provides a more comprehensive overview of the laru microbiota.

Identification and Fermentation Characteristics of Lactic Acid Bacteria Isolated from Dongchimi as Starter for Radish Juice (무 쥬스 제조를 위한 starter로써 동치미에서 분리한 유산균의 동정 및 발효 특성)

  • Kim, Jung-Hee;Kim, Jong-Il
    • Korean Journal of Microbiology
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    • v.35 no.4
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    • pp.307-314
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    • 1999
  • This study was aimed at the investigation of the possibility of the addition of lactic acid bacteria as "starter"for the preparation of radish juice. Forty strains of lactic acid bacteria were isolated from dongchimi that was fermented by a traditional method. The isolates were assorted into 5groups, Leuconostoc mesenteroides subsp. mesenteroides (J-9), Lacobacillus brevis (J-12), Lactobacillus fermentum (J-7), Lactobacillus sake (J-20), and Lactobacillus plantarum (J-39). Leuconostoc mesenteroides was predominated in the sample of dongchimi with frequency of 52.5%. Each of the strain, which exhibited the beat growth in the species, was selected in the 5species, and investigation of the fermentation characteristcis was carried out. The fermentation were performed for 9 days at 25${\circ}C$ after the inoculation of 0.3% ($10^{6}$ cfu/㎖) to each ultra-filtrated radish juice. The pH, total acidity, content of non-volatile organic acids were examined during the fermentation period. Lactobacillus plantarum showed the highest growth rate and the growth rate of Lactobacillus sake was the lowest. The pH (6.3-6.36) and total acidity (0.09-1.0 %) fo the ultrafiltrated radish juice before fermentation were changed to 3.2-4.3 and 0.65-1.2% after 9days, respectively. The changes of the pH and total acidity were related with the growth of the lactic acid bacteria; the better growth of lactic acid bacteria, the more rapid decrease of pH and increase of the total acidity. when the amount of non-volatile organic acids were estimated during fermentation, citric acid, malic acid, malonic acid, and succinic acid were decreased in all cases. However, the content of lactic acid increased with the progression of fermentation. L. mesenteroides (J-9), L. brevis (J-12) and L. fermentum (J-7) were chosen for the candidates of the starter for the lactic fermentation of radish juice based on the biochemical analysis and sensory evaluation.

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Screening and Identification of Soy Curd-Producing Lactic Acid Bacteria (두유 커드를 생산하는 김치 유래 젖산균의 동정)

  • Kim, Ro-Ui;Ahn, Soon-Cheol;Yu, Sun-Nyoung;Kim, Kwang-Youn;Seong, Jong-Hwan;Lee, Young-Guen;Kim, Han-Soo;Kim, Dong-Seob
    • Journal of Life Science
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    • v.21 no.2
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    • pp.235-241
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    • 2011
  • The purpose of this study was to isolate soy curd forming bacterial strains. Soy curd forming bacteria were isolated from Kimchi, a traditional Korean vegetable food that is fermented using lactic acid bacteria. Among 196 bacterial strains, ten isolates (strain No. 2-2-2, 2-15-2, 2-18-1, 2-19-2, 3-4-1, 3-4-2, 3-8-1, 3-8-3, 3-17-1, 4-39-5) formed firm soy curd. The isolated bacterial strains were identified by molecular biological and biochemical analyses. The genomic DNAs extracted from the isolated bacterial strains were used as a template for PCR amplification of 16S rDNA region. By comparing the results of the 16s rDNA sequences with GenBank data, the isolated strains were identified as Leuconostoc mesenteroides group and Lactobacillus sakei group. The phylogenetic position of soy curd forming strains and their related taxa were investigated using neighbor-joining method. L. mesenteroides group was further identified as L. mesenteroides subsp. dextranicum based on biochemical properties. L. sakei group was named Lactobacillus sp., because it showed a variety of biochemical properties.