• Title/Summary/Keyword: Lactobacillus species

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Development of Species-Specific Primers for PCR Identification of Lactobacillus hilgardii and Lactobacillus farciminis in Kimchi

  • Lee, Myung-Ki;Ku, Kyung-Hyung;Kim, Young-Jin;Kim, Kyung-Hee;Kim, Yu-Ri;Yang, Hye-Jung
    • Preventive Nutrition and Food Science
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    • v.15 no.2
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    • pp.159-166
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    • 2010
  • The aim of this study was to develop species-specific primer sets for kimchi Lactobacillus. Known gene sequences of Lactobacillus 16S rRNA were collected from the NCBI Gene bank, and 69 primer sets were designed using the homologous gene sequence. Six species of kimchi Lactobacilli were used as reference strains: Lactobacillus brevis KCTC3102, Lactobacillus farciminis KCTC3681, Lactobacillus fermentum KCTC3112, Lactobacillus hilgardii KCTC3500, Lactobacillus plantarum KCTC3099, and Lactobacillus sanfranciscensis KCTC3205. PCR amplification and gel electrophoresis were performed to identify the accuracy and specificity of the developed primer set. The results show that the primer set of 5'-aagcctgcgaaggcaag-3' & 5'-aggccaccggctttg-3', 5'-acatactatgcaaatctaagagattagacg-3' & 5'-actgagaatggctttaagagattagcttac-3' resulted in a specific PCR band on L. hilgardii, and primer set of 5'-ctaataccgcataacaactactttcacat-3' & 5'-aacttaataaaccgcctacattctctttac-3' on L. farciminis. The results indicate that the developed primer sets can provide a useful tool for the identification and differentiation of L. hilgardii and L. farciminis from other Lactobacillus species of kimchi.

Analysis of Lactobacillus Species Isolated from the Vagina of Korean Women (한국여성의 질내 Lactobacillus 균종분포)

  • Kim, Kack-Kyun;Kook, Yoon-Hoh;Chang, Woo-Hyun
    • The Journal of the Korean Society for Microbiology
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    • v.16 no.1
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    • pp.35-38
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    • 1981
  • Lactobacillus species isolated from posterior fornix of the vagina of 410 Korean women, who visited several local clinics and the Family Health Clinic of the Institute of Reproductive Medicine and Population, Seoul National University in Seoul were identified by their physiological and biochemical characteristics. And the results were as follows. 1) Of the 410 women 297(72.4%) gave the positive culture for Lactobacillus. 2) Isolated 200 strains were differentiated into following species. L. salivarius(43 strains, 18.7%) and L. leichmannii(42, 18.3%) were the most common species followed by L. plantarum(34, 14.8%) L.brevis(22, 9.6%), L. fermenti(19, 8.3%), L. cellobiosus(15, 6.5%), L. acidophilus(14, 6.1%), L. delbrueckii(10, 4.3%) and L. casei(1, 0.4%) and there were 30 strains(13%) which could not have been identified by the methods employed in this study. 3) Age seems to have no influence on the distribution of Lactobacillus species in vagina and it seems like that the overall distribution of Lactobacillus species in vagina does not change during the active reproductive period of women.

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Reevaluation of Isolation and Identification of Gram-positive Bacteria in Kimchi (김치에 서식하는 Gram 양성세균의 분리 및 동정의 재평가)

  • 임종락;박현근;한홍의
    • Korean Journal of Microbiology
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    • v.27 no.4
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    • pp.404-414
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    • 1989
  • Attempts were made to isolate and identify Gram-positive or lactic acid bacteria in Kimchi fermentation. Species diversity depended on isolation media and temperatures, and diversity tended to be reduced with decrease of temperature. MRS and KM (natural medium prepared from Kimchi materials) were suitable respectively for isolation and present number of species. Identification of isolates was performed by dichotomous identification schemes arranged on the basis of Bergey's manual of Systematic Bacteriology (1986). Gram-positive bacteria isolated at different temperatures (5, 15, $25^{\circ}C$) were 5 species of Leuconostoc, 4 species of Streptococcus, 3 species of Pediococcus, 2 species of Bacillus and 18 species of Lactobacillus. Species with high frequency of appearance were Lactobacillus plantarum, Streptococcus raffinolactis, Leuconostoc mesenteroides subsp. mesenteroides at $25^{\circ}C$, L. plantarum, Lactobacillus fructosus, L. mesenteroides subsp. mesenteroides at $15^{\circ}C$ and L. mesenteroides subsp. mesenteroides, Leuconosotoc paramesenteroides, Lactobacillus maltaromicus at $15^{\circ}C$. In general, Kimchi fermentation was achieved by Lactobacillus spp. (59.7% frequency) at $25^{\circ}C$ and Leuconostoc spp. (65.2% frequency) at $5^{\circ}C$. Pediococcus cerevisiae and Streptococcus faecalis which have been so far known as bacteria of Kimchi fermentation were not isolated.

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Isolation of Leuconostoc and Weissella Species Inhibiting the Growth of Lactobacillus sakei from Kimchi (김치로부터 Lactobacillus sakei 생육저해 Leuconostoc 및 Weissella 속 균주의 분리)

  • Lee, Kwang-Hee;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.39 no.2
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    • pp.175-181
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    • 2011
  • Kimchi is a group of traditional fermented vegetable foods in Korea and known to be the product of a natural mixed-fermentation process carried out principally by lactic acid bacteria (LAB). According to microbial results based on conventional identification, Leuconostoc mesenteroides and Lactobacillus plantarum were considered to be responsible for the good taste and over-ripening of kimchi, respectively. However, with the application of phylogenetic identification, based on 16S ribosomal RNA gene similarities, a variety of Leuconostoc and Lactobacillus species not detected in the previous studies have been isolated, together with a species in the genus Weissella. Additionally, Lactobacillus sakei has been accepted as the most populous LAB in over-ripened kimchi. In this study, Leuconostoc and Weissella species inhibiting the growth of Lb. sakei were isolated from kimchi for future applications to do with kimchi fermentation. From 25 kimchi samples, 378 strains in the genera Leuconostoc and Weissella were isolated and 68 strains identified as Lc. mesenteroides, Lc. citreum, Lc. lactis, W. cibaria, W. confusa, and W. paramesenteroides exhibited growth inhibition against Lb. sakei. Most of the strains also had antagonistic activities against Lb. brevis, Lb. curvatus, Lb. paraplantarum, Lb. pentosus, and Lb. plantarum. Their antagonistic activities against Lb. sakei were more remarkable at lower temperatures of incubation.

PCR-Based Detection of Lactic Acid Bacteria in Korean Fermented Vegetables with recA Gene Targeted Species-Specific Primers (RecA 유전자 특이적 PCR을 이용한 전통 침채류 유래 유산균의 검출)

  • Shim, Sang-Min;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.36 no.2
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    • pp.96-100
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    • 2008
  • Diversity of lactic acid bacteria involved in 5 Korean fermented vegetables (Cot kimchi, Dongchimi, Baechu kimchi, Oisobagi, and Chonggak kimchi) was investigated using PCR-based method. PCR primer pairs targeted the recA gene were used for the detection of 7 species of lactic acid bacteria mainly found in kimchi and Lactobacillus acidophilus involved in dairy fermentation. Lactobacillus plantarum and Lactobacillus sakei were detected in all samples tested but Lactobacillus paraplantarum, Lactobacillus pentosus, and Lb. acidophilus were not detected. Lactobacillus brevis and Leuconostoc citreum were detected only from Baechu kimchi and Leuconostoc mesenteroides was detected from Got kimchi, Dongchimi, Baechu kimchi, and Oisobagi. The difference of detected species from fermented vegetables may be originated from the difference of main materials. Lb. plantarum and Lb. sakei are supposed to be broadly involved in Korean fermented vegetables.

Real-Time PCR Monitoring of Lactobacillus sake, Lactobacillus plantarum, and Lactobacillus paraplantarum during Kimchi Fermentation

  • Um, Sang-Hee;Shin, Weon-Sun;Lee, Jong-Hoon
    • Food Science and Biotechnology
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    • v.15 no.4
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    • pp.595-598
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    • 2006
  • Semi-quantitative monitoring of Lactobacillus sake and Lactobacillus plantarum, major and minor microorganisms in kimchi, respectively, and Lactobacillus paraplantarum, recently shown to be present in kimchi, was carried out by real-time polymerase chain reaction (PCR). Changes in the 3 species during kimchi fermentation were monitored by the threshold cycle ($C_T$) of real-time PCR. As fermentation proceeded at $15^{\circ}C$, the number of L. sake increased dramatically compared to those of L. plantarum and L. paraplantarum. During fermentation at $4^{\circ}C$, the growth rates of the 3 species decreased, but the proportions of L. plantarum and L. paraplantarum in the microbial ecosystem were almost constant. Considering the $C_T$ values of the first samples and the change in the $C_T$ value, the number of L. sake is no doubt greater than those of L. plantarum and L. paraplantarum in the kimchi ecosystem. L. sake seems to be one of the major microorganisms involved in kimchi fermentation, but there is insufficient evidence to suggest that L. plantarum is the primary acidifying bacterium.

Effect of Low Temperature on the Qualities of Long-term Fermented Kimchi (Korean Pickled Cabbage) (저온이 묵은지의 발효에 미치는 영향)

  • Ji, Seol-Hee;Han, Woo-Cheul;Lee, Jae-Cheol;Cheong, Chul;Kang, Soon-Ah;Lee, Je-Hyuk;Jang, Ki-Hyo
    • Food Science and Preservation
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    • v.16 no.6
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    • pp.804-809
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    • 2009
  • Recently, many Koreans have started to consume aged kimchi fermented long-term at low temperatures. In the present study, the effect of low temperature ($5^{\circ}C$) on pH, titratable acidity (TA), organic acid level, viable microbial cell count, amino acid concentration, and polygalacturonase activity (PG) during long-term fermentation (46 weeks) of kimchi, were evaluated. After 10 weeks of fermentation, kimchi had a pH of 4.1 and a TA of 1.0%, respectively after 46 weeks fermentation, these values were 3.9 and 1.3%, respectively. Lactic acid, the ratio of lactic acid to acetic acid, and the ratio of Lactobacillus species/Leuconostoc species in kimchi increased as fermentation progressed from 10 weeks to 46 weeks. However, total viable cell counts of aerobic bacteria, yeasts, Lactobacillus species, and Leuconostoc species, free amino acid levels, and PG decreased as the fermentation period was extended from 10 weeks to 46 weeks.

Comparative Tests on the Acid Tolerance of Some Lactic-Acid-Bacteria Species Isolated from Lactic Fermented Products (젖산 발효제품에서 분리한 유산균의 내산성 비교)

  • Sim, Jae-Hun;Oh, Se-Jong;Kim, Sang-Kyo;Baek, Young-Jin
    • Korean Journal of Food Science and Technology
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    • v.27 no.1
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    • pp.101-104
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    • 1995
  • We isolated sixty lactic acid bacteria(LAB) from lactic fermented products. Among 60 isolates of LAB, 30 isolates were identified as Lactobacillus casei ssp.(5 strains), Lactobacillus acidophilus(2 strains), Lactobacillus delbrueckii ssp. bulgaricus(6 strains), Lactobacillus plantarum(4 strains), Streptococcus salivarius ssp. thermophilus(11 strains), and Streptococcus faecalis(2 strains). The acid tolerance and bile resistance of 30 LAB were determined. Because the acid tolerance was affected by the initial cell concentrations, the analysis of covariance could be used to remove the effect of initial cells on acid tolerance when testing for differences in acid tolerance among six species. Viability of LAB under acidic condition, pH 3 for 2 hours at $37^{\circ}C$, was significantly different among the species. L. casei and L. acidophilus strains showed great viability, but L. bulgaricus and S. thermophilus strains were very weak in acid tolerance.

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Antioxidant Activity and Tolerance to Reactive Oxygen Species of Lactobacillus spp. (Lactobacill spp. 의 황산화 효과 및 활성산소에 대한 내성)

  • Kim, H.S.;Jeong, S.G.;Chae, H.S.;Ham, J.S.;Ahn, C.N.;Lee, J.M.
    • Journal of Animal Science and Technology
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    • v.46 no.6
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    • pp.1007-1012
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    • 2004
  • The four Lactobacillus spp. were investigated for their antioxidant properties, including antioxidant activity, tolerance to reactive oxygen species, hydroxy radical scavenging activity and ferrous iron che1ating activity. Also, activities of superoxide dismutase and glutathione peroxidase were investigated. From the results of this work, the intact cell and cell lysate of L. casei KCTC 3260 were exhibited highest antioxidant activity. L. casei KCTC 3260 also showed strong tolerance to reactive oxygen species. This strain showed highest glutathione peroxidase activity among the tested strains.

Origin of lactic acid bacteria in mulkimchi fermentation

  • Hwang, Chung Eun;Haque, Md. Azizul;Hong, Su Young;Kim, Su Cheol;Cho, Kye Man
    • Journal of Applied Biological Chemistry
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    • v.62 no.4
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    • pp.441-446
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    • 2019
  • The assortment of endophytic lactic acid bacteria (LAB) in kimchi derives from its raw vegetables, which include Chinese cabbage, radish, welsh onion, onion, garlic, red pepper, and ginger. These vegetables were examined during mulkimchi fermentation using gene-specific multiplex polymerase chain reaction and 16S ribosomal RNA sequence analysis. Sixteen species from five LAB genera (Leuconostoc, Lactobacillus, Lactococcus, Pediococcus, and Weissella) appeared in the raw kimchi materials. Interestingly, nine LAB species were identified in mulkimchi on fermentation day 0 as follows: Leuconostoc carnosum, Leuconostoc citreum, Leuconostoc gelidum, Leuconostoc inhae, Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus sakei, Lactococcus lactis, and Weissella confusa. Seven additional LAB species were present in mulkimchi at fermentation day 9 as follows: Leuconostoc gasicomitatum, Leuconostoc kimchii, Lactobacillus brevis, Lactobacillus curvatus, Lactobacillus pentosus, Pediococcus pentosaceus, and Weissella koreensis. These species corresponded completely with the LAB in kimchi vegetables. Wei. confusa was the predominant LAB during early fermentation (pH 6.20 to 4.98 and acidity 0.20 to 0.64%), while Lac. sakei, Lac. plantarum, and Wei. koreensis became dominant later in fermentation (pH 4.98 to 3.88 and acidity 0.64 to 1.26%). These results collectively demonstrate that the LAB involved in mulkimchi fermentation originates from the raw vegetables examined.