• Title/Summary/Keyword: Lactobacillus identification

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Isolation and Identification of Lactobacillus sp. Produced r-Aminobutyric Acid(GABA) from Traditional Salt Fermented Anchovy (멸치 젓갈로부터 r-Aminobutyric Acid(GABA)를 생성하는 Lactobacillus 속의 분리.동정)

  • 전재호;김현대;이홍수;류병호
    • The Korean Journal of Food And Nutrition
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    • v.17 no.1
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    • pp.72-79
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    • 2004
  • This study was conducted to investigate the identification of lactic acid bacteria produced ν-aminobutyric acid(GABA) from traditional salt fermented anchovy. There was no appreciable difference in the number of lactic acid bacteria from fermented anchovy. Among the types of lactic acid bacteria, three strains of lactic acid bacteria produced ν-aminobutyric acid from those sample were identified temporary as name of Lactobacillus brevis BH-21, Lactobacillus rhamnosus BH-32 and Lactobacillus plantarum BH-38 by using gram positive identification(GPI) card and API 50 kit, respectively. 3 strains of Lactobacillus sp. were found to produce GAB A in the culture of filtrate. Lactobacillus brevis BH-21 produced GABA, some of which yielded 43.2 mg/mL GABA in the medium of 0.1% glucose, 0.1% yeast extract, 0.05% polypeptone, 0.002% MgSO$_4$$.$4H$_2$O, 0.001% FeSO$_4$$.$7H$_2$O, 0.01% NaCl, 0.1% monosodium glutamate, pH 6.0. This result suggests that Lactobacillus brevis BH-21 has the potential to be developed as a strain of GABA production.

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.

Identification of Lactobacillus spp. associated with nematodes in peach farm soil (복숭아 농장 토양에서 Nematodes와 연관된 Lactobacillus spp.의 분리 및 동정)

  • Lee, Woo-Hyun;Choi, Jae Im;Lee, Jin Il;Lee, Won-Pyo;Yoon, Sung-Sik
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.163-169
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    • 2017
  • Strains D4 and D5 were isolated from peach-rotten soil during the peach harvest season. The isolates were identified based on morphological and biochemical characterization, and identification was determined by 16S rRNA gene sequencing. Results showed that D4 has high similarity to Lactobacillus plantarum ATCC $14917^T$ and Lactobacillus pentosus ATCC $8041^T$ at 99.05% and 98.98%, respectively. D5 was also similar to Lactobacillus pentosus ATCC $8041^T$ and Lactobacillus plantarum ATCC $14917^T$ at 98.71% and 98.64%, respectively. In contrast, isolates showed differences in carbohydrate utilization in comparison to Lactobacillus plantarum ATCC $14917^T$ and Lactobacillus pentosus ATCC $8041^T$. In view of this we performed VITEK MS matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis, multiplex PCR fingerprinting, and random amplified polymorphic DNA (RAPD)-PCR to further confirm the identification of D4 and D5. The results of these analyses showed that both strains were most similar to Lactobacillus plantarum.

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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Genetic Identification of the Kimchi Strain Using PCR-based PepN and 16S rRNA Gene Sequence (PepN과 16S rRNA Gene Sequence 및 PCR 방법을 이용한 김치 젖산균의 동정)

  • Lee, Myung-Ki;Park, Wan-Soo;Lee, Byong-H.
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1331-1335
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    • 2000
  • The WL6 strain isolated from Kimchi could not be made scientific name because it was identified as three species, i.e., Leuconostoc mesenternides ssp cremoris, Leu. mesenteroides ssp. dextranicum or Lactobacillus bifermentans when it was tested by API kit or Biolog system methods. The unidentifiable WL6 strain was finally reclassified as Lactobacillus bifermentans by genetic identification using two PCR-based specific sequence primer sets which were originated from homologous pepN and 16S rRNA genes.

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Rapid Identification of Lactobacillus and Bifidobacterium in Probiotic Products Using Multiplex PCR

  • Sul, Su-Yeon;Kim, Hyun-Joong;Kim, Tae-Woon;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.17 no.3
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    • pp.490-495
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    • 2007
  • Lactic acid bacteria (LAB) are beneficial for the gastrointestinal tract and reinforce immunity in human health. Recently, many functional products using the lactic acid bacteria have been developed. Among these LAB, Lactobacillus acidophilus, Lactobacillus rhamnosus, Bifidobacterium longum, and Bifidobacterium bifidum are frequently used for probiotic products. In order to monitor these LAB in commercial probiotic products, a multiplex PCR method was developed. We designed four species-specific primer pairs for multiplex PCR from the 16S rRNA, 16S-23S rRNA intergenic spacer region, and 23S rRNA genes in Lactobacillus acidophilus, Lactobacillus rhamnosus, Bifidobacterium longum, and Bifidobacterium bifidum. Using these primer pairs, 4 different LAB were detected with high specificity in functional foods. We suggest that the multiplex PCR method developed in this study would be an efficient tool for simple, rapid, and reliable identification of LAB used as probiotic strains.

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.

Isolation and Identification of Lactobacilli from Fermented Sausages (발효소시지로부터 유산생성균의 분리 및 동정)

  • 고명수;이명섭;김창한
    • Microbiology and Biotechnology Letters
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    • v.22 no.5
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    • pp.544-549
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    • 1994
  • Lactobacilli proliferating in fermented sausages of the specific ripening conditions were isolated from fermented sausages, manufactured in the absence of an added starter, during ripening under controlled temperature-humidity conditions. Based on morphological, physiological and bio- chemical characteristics and carbohydrate fermentation of isolated strains, three strains of isolates were identified as Lactobacillus curvatus, two strains as Lactobacillus sake. Optimal temperature and pH for growth of isolated strains were 30$\circ$C and pH 6.0~7.0, respectively. These strains were salt tolerant, multiplying in the presense of 6~8% NaCl.

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Identification and Characteristics of Lactic Acid Bacteria Isolated from Shellfishes (패류로부터 젖산 세균의 분리 및 특성)

  • Kang, Chang-Ho;Jeong, Ho-Geon;Koo, Ja-Ryong;Jeon, Eun-Jin;Kwak, Dae-Yung;Hong, Chae-Hwan;Kim, Si-Hwan;Seo, Ji-Yeon;Han, Do-Suck;So, Jae-Seong
    • KSBB Journal
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    • v.27 no.3
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    • pp.151-156
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    • 2012
  • Lactic acid is an important product arising from the anaerobic fermentation by lactic acid bacteria (LAB). It is used in the pharmaceutical, cosmetic, chemical, and food industries as well as for biodegradable polymer and green solvent production. The poly lactic acid (PLA) is an important material for bio-plastic manufacturing process. For PLA production by new LAB, we screened LAB isolates from shellfish. A total of 28 LAB were isolated from various shellfishes. They were all Gram positive, oxidase and catalase negative. Based on API 50CHL kit, 7 strains among the 28 isolates were identified as Lactobacillus plantarum, 6 strains as Lactobacillus delbrueckii, 5 strains as Leuconostoc mesenteroides, 3 strains as Lactobacillus brevis, 2 strains as Lactococcus lactis, 1 strain as Lactobacillus salivarius, 1 strain as Lactobacillus paracasei, 1 strain as Lactobacillus pentosus, 1 strain as Lactobacillus fermentum and 1 strain as Pediococcus pentosaceu. Also, we examined the amount of total lactic acid produced by these new strains by HPLC analysis with Chiralpak MA column. One strain E-3 from Mytilus edulis was indentified as Lactobacillus plantarum and found to produce 20.0 g/L of D-form lactic acid from 20 g/L of dextrose. Further studies are underway to increase the D-lactic acid production by E-3.

Isolation and Identification of Lactic Acid Bacteria from Commercial Kimchi (시판김치로부터 젖산균의 분리 및 동정)

  • Ko, Jung-Lim;Oh, Chang-Kyung;Oh, Myung-Cheol;Kim, Soo-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.6
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    • pp.732-741
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    • 2009
  • This study was carried out to identify lactic acid bacteria isolated from commercial Kimchi. Twelve lactic acid bacteria strains were isolated from Chinese cabbage kimchi (Baechu kimchi) that was fermented for 4 days at room temperature after making kimchi, 6 strains from pickled ponytail radishes (Chongkak kimchi) that was fermented for 2 days, and 15 strains in radish cube kimchi (Kaktugi) that was fermented for 5 days, and 23 strains were isolated in pickled Wakegi (Pa kimchi) that was fermented for 4 days. Eight strains among the lactic acid bacteria of 12 strains isolated from Baechu kimchi (pH 4.0) were identified as Lactobacillus plantarum, 1 strain as Leuconostoc lactis, 2 strains as Lactobacillus casei subsp. pseudoplantarum, and 1 strain as Lactobacillus sake. Three strains among the lactic acid bacteria of 6 strains isolated from Chongkak kimchi (pH 4.5) were identified as Leuconostoc paramesenteroides, 2 strains as Leuconostoc mesenteroides subsp. mesenteroides, and 1 strain as Lactobacillus plantarum. Two strains among the 15 strains isolated in Kaktugi (pH 4.0) were identified as Leuconostoc lactis, 3 strains as Leuconostoc mesenteroides subsp dextranicum, 4 strains as Lactobacillus casei subsp. pseudoplantarum, and 4 strains as Lactobacillus coryniformis subsp. torquens. Twenty-two strains among the 23 strains isolated from Pa kimchi (pH 4.1) identified as L. plantarum and 1 strain was as Lactobacillus sake. From the results above, the dominant species of Baechu kimchi was confirmed as L. plantarum, Chongkak kimchi as L. paramesenteroides, Kaktugi as L. casei subsp. pseudoplantarum and L. coryniformis subsp. torquens, and Pa kimchi as L. plantarum.