• Title/Summary/Keyword: rDNA ITS sequence

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Characteristics and Partial Purification of a Bacteriocin Produced by Pediococcus damnosus JNU 534 (Pediococcus damnosus JNU 534가 생산하는 박테리오신의 특성 및 정제)

  • Lee, Jae-Won;Han, Su-Min;Yun, Bo-Hyun;Oh, Se-Jong
    • Food Science of Animal Resources
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    • v.31 no.6
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    • pp.952-959
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    • 2011
  • A new bacteriocin-producing lactic acid bacteria (LAB) which has been isolated from kimchi was identified as Pediococcus damnosus by use of API kit and 16S rDNA sequencing, and designated as P. damnosus JNU 534. The bacteriocin produced by P. damnosus JNU 534 markedly inhibited the growth of some of LAB and Listeria monocytogenes, whereas other pathogens including Gram negative bacteria were not susceptible. The production of bacteriocin started at the beginning of exponential phase and reached maximum activity at the early stationary phase. The bacteriocin was stable on the wide pH range of 2-9 and heat treatment up to $100^{\circ}C$ for 15 min. The antimicrobial compound was inactivated by treatments of proteolytic enzymes indicating its proteinaceous in nature. The bacteriocin was purified by 30% ammonium sulfate precipitation followed by hydrophobic interaction column and $C_{18}$ column chromatography. The estimated molecular weight of the bacteriocin using tricine SDS-PAGE was approximately 3.4 kDa and the identified N-terminal amino acid sequence was $NH_2$-ILLEELNV.

Physiological Characteristics and GABA Production of Lactobacillus acidophilus RMK567 Isolated from Raw Milk (원유에서 분리한 Lactobacillus acidophilus RMK567의 GABA 생성 및 생리적 특성)

  • Lim, Sang-Dong;Kim, Kee-Sung;Do, Jeong-Ryong
    • Food Science of Animal Resources
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    • v.29 no.1
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    • pp.15-23
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    • 2009
  • In order to develop a new starter for fermented milk, 2082 bacteria were isolated from raw milk. The strain that showed excellent acid forming and ${\gamma}$-aminobutyric acid (GABA) production ($711.40{\mu}g/g$ D.W) characteristics after incubation at $37^{\circ}C$ for 18 hr was selected and identified as Lactobacillus acidophilus by the result of API carbohydrate fermentation pattern and 16S rDNA sequence. L. acidophilus RMK567 was investigated for its physiological characteristics. RMK67 strain showed good GABA production compared with commercial lactic acid bacteria. The optimum growth temperature of L. acidophilus RMK567 was $40^{\circ}C$ and cultures took 15 hr to reach pH 4.3. L. acidophilus RMK567 showed higher sensitivity to penicillin-G, novobiocin, as compared to other 14 different antibiotics. However, it showed more resistance to kanamycin, neomycin, streptomycin. It showed higher leucine arylamidase and ${\beta}$-galactosidase activities compared to 16 other enzymes. It was comparatively tolerant to bile juice and able to survive at pH 2 for 3 hr. It showed resistence to Escherichia coli, Salmonella typhimurium and Staphylococcus aureus with rates of 29.2%, 39.1% and 51.4%, respectively. Based on these and previous results, L. acidophilus RMK567 could be an excellent starter culture for fermented milk with excellent GABA contents.

Occurrence of Leaf Mold Pathogen Fulvia fulva Isolates Infecting Tomato Cf-9 Cultivars in Korea (토마토 Cf-9 저항성 품종에 잎곰팡이병을 일으키는 Fulvia fulva 균주의 국내 발생)

  • Lee, Ji Hyun;Park, Myung Soo;Kim, Jin-Cheol;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Heung Tae;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.31 no.6
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    • pp.740-747
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    • 2013
  • Leaf mold symptoms were found on commercial tomato cultivars carrying the Cf-9, a resistance gene to leaf mold pathogen Fulvia fulva in 2012 at Buyeo, Chungnam in Korea. Fifteen-fungal isolates were obtained from four Cf-9 cultivars of tomato including 'Cutie', 'otaerangdia', 'Unicorn' and 'Rapito'. Due to their same morphological appearances and colony color, nine isolates were selected and identified as F. fulva based on molecular analysis of the internal transcribed spacer rDNA sequence. Pathogenicity of the 15 isolates on five commercial cultivars carrying Cf-4, Cf-5, and Cf-9 were tested. All the isolates showed strong pathogenicity on Cf-9 cultivars, 'Cutie' and 'Dotaerangdia', and Cf-5 cultivar, 'Yoyocaptain'. In contrast, on Cf-4 cultivar, 'Superdotaerang', five isolates were virulent and the other isolates were not. In addition, two fungal isolates, infecting Cf-9 cultivar and non-infecting Cf-4 cultivar, were selected and their pathogenicity was tested on 17 commercial cultivars reported as tomato having Cf-9 resistance gene. Among them, 15 cultivars were susceptible and 2 cultivars were resistant. It is likely that the two cultivars include other resistance gene. To our knowledge, this is the first report on the occurrence of Cf-9 infecting F. fulfva strains in Korea.

Characteristics of Lactic Acid Bacteria Isolated from Kefir Made of Goat Milk (산양유 Kefir 발효물에서 분리한 유산균의 특성)

  • Lim, Young-Soon;Kim, Soo-Young;Lee, Si-Kyung
    • Food Science of Animal Resources
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    • v.28 no.1
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    • pp.82-90
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    • 2008
  • Two strains of pure lactic acid bacteria capable of forming both acid and slime were isolated from the kefir made of goat milk. The isolated strains observed by morphological and physiological properties, and their 16S rDNA partial sequence were identified as Streptococcus salivarius subsp. thermophilus(LFG-1) and Lactococcus lactis subsp. lacits(LFG-2) with over 99% homology. The optimum temperature of Str. salivarius subs. thermophilus LFG-1 for growth was $40-45^{\circ}C$, and its generation time was 40.6 minutes. The final pH of cultured broth by Str. salivarius subsp. thermophilus LFG-1 and the commercial strain Str. thermophilus Body-1 for 24hr at $37^{\circ}C$ were 4.30 and 4.55, respectively. The coagulative activity of Str. salivarius subsp. thermophilus LFG-1 was almost as strong as that of commercial strain Str. thermophilus Body-1. However, the LFG-2 strain showed lower coagulative activity than Str. thermophilus Body-1. The survival rate of lactic acid bacteria were between 22-29% in 0.3% bile extract. At pH 1.0 all of the bacteria were killed, and most of lactic acid bacteria died against pH 3.0. However, all lactic acid bacteria survived well at pH 4.5.

Isolation and Characteristics of Alginate-Degrading Methylobacterium sp. HJM27 (알긴산 분해 Methylobacterium sp. HJM27 균주의 분리 및 특성)

  • Kim, Ok-Ju;Lee, Dong-Geun;Lee, Sung-Mok;Lee, Suck-June;Do, Hyung-Joo;Park, Hye-Jin;Kim, Andre;Lee, Jae-Hwa;Ha, Jong-Myung
    • Microbiology and Biotechnology Letters
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    • v.38 no.2
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    • pp.144-150
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    • 2010
  • This study was aimed to screen bacteria of high alginate-degrading activity, to select the nitrogen source and concentration of NaCl and sodium alginate for the production of alginate-degrading enzyme, and to determine reaction conditions of enzyme. A novel alginate-degrading bacterium was isolated from abalone (Haliotis discus hannai) and named Methylobacterium sp. HJM27 by 16S rDNA sequence analysis. The optimum culture conditions for the production of alginate-degrading enzyme were 1.0% sodium alginate, 0.5% peptone, 0.3% yeast extract, 1.5% NaCl, $25^{\circ}C$ and 48 hours incubation time. The raw enzyme showed the highest activity at $25^{\circ}C$ and pH 9, and produced 1.217 g - reducing sugar per liter in 0.8% (w/v) sodium alginate for 30 minutes. Methylobacterium sp. HJM27 and its alginate-degrading enzyme would be useful for the production of bioenergy and biofunctional oligosaccharides from seaweed.

Isolation of an Agarase-producing Persicobacter sp. DH-3 and Characterization of its β-agarase (Agarase를 생산하는 Persicobacter sp. DH-3의 분리 및 β-agarase의 특성)

  • Heo, Da-Hye;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.29 no.2
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    • pp.158-163
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    • 2019
  • The purpose of this study was to isolate a new marine agarase-producing bacterium. Agarase can hydrolyze agar and agarose to produce agarooligosaccharides or neoagarooligosaccharides, which possess many physiological functions. Strain DH-3 was isolated from seawater collected from the coast of Yeosu at Jeollanam province, Korea. A 16S rDNA sequence analysis showed this strain to be Persicobacter sp. DH-3. Extracellular agarase was prepared from culture media of Persicobacter sp. DH-3 and used for characterization. Relative activities at 20, 30, 40, 50, 60, and $70^{\circ}C$ were 50, 55, 70, 100, 90, and 50%, respectively. Relative activities at pH 5, 6, 7, and 8 were 75, 100, 90, and 75%, respectively. The enzyme showed maximum activity at $50^{\circ}C$ in a 20 mM Tris-HCl buffer at pH 6. This enzyme could be useful, as agar is in liquid state at $50^{\circ}C$. Agarase activities were maintained at 80% or more for 2 hr at 20, 30, and $40^{\circ}C$. Thin layer chromatography analysis suggested that Persicobacter sp. DH-3 produced extracellular ${\beta}$-agarases as it hydrolyzed agarose to produce neoagarohexaose and neoagarotetraose. In addition, zymogram analysis confirmed that Persicobacter sp. DH-3 produces at least three agar-degrading enzymes with molecular weights of 45, 70, and 140 kDa. Therefore, it is expected that agarases from Persicobacter sp. DH-3 could be used to produce functional neoagarooligosaccharides.

Distribution and Species Diversity of Wild Yeasts Isolated from Flowers in Korea (국내에 서식하는 꽃에서 분리한 야생 효모 분포 및 종 다양성)

  • Kim, Jeong-Seon;Lee, Miran;Kim, Jae Yoon;Heo, jun;Kwon, Soon-Wo;Yun, Bong Sik;Kim, Soo-Jin
    • The Korean Journal of Mycology
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    • v.48 no.4
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    • pp.475-484
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    • 2020
  • Various indigenous yeasts were isolated and obtained from flowers in the Republic of Korea, and their distribution and species diversity were studied. Seventy-seven flowers were collected from 25 areas in Korea, and 502 yeast strains were isolated from these flowers. A total of 50 species were identified by comparing large subunit rDNA gene sequence homology with the type strains of yeasts. The analysis of yeast distribution showed that the dominant yeast species were Aureobasidium pullulans, A. leucospermi, and Filobasidium magnum in each region and flower samples. Except for the above three yeast species, no species of yeasts showed any meaningful distribution among the habitat regions and sources. In conclusion, 50 species of indigenous yeasts were obtained from flowers that can be used as industrial resources, and the data could be used for further research on yeast diversity and interactions between yeast and its host.

Occurrence of Soft Rot on Raspberry (Rubus crataegifolius) Caused by Rhizopus oryzae in Korea (Rhizopus oryzae에 의한 산딸기 무름병)

  • Kwon, Jin-Hyeuk;Kang, Dong-Wan;An, Jae-Uk;Choi, Ok-Hee;Kwak, Youn-Sig
    • Research in Plant Disease
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    • v.17 no.3
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    • pp.396-398
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    • 2011
  • Soft rot disease on Raspberry (Rubus crataegifolius Bunge) was observed in sale boxes at Jinju City Wholesale Market of Agricultural Products in June 2010. The infected fruits were rapidly water-soaked, softened, black and eventually rotted. The colonies on the infected fruits were white to light brown, formed numerous sporangiospores. Optimum temperature for the mycelial growth of the causal fungus on PDA was $30^{\circ}C$ and growth was still apparent at $37^{\circ}C$. Sporangia were globose, white at early and gradually to black, and 40-210 ${\mu}m$ in diameter. Sporangiophores were white to mid brown as maturation and 8-20 ${\mu}m$ in diameter. Columella were globose to sub-globose, and the size of the diameter was 85-120 ${\mu}m$ in diameter. Sporangiospores were sub-globose, rhomboidal and irregular, streaked and 5-10 ${\mu}m$ in length. On the basis of symptom, mycological characteristics, ITS rDNA sequence analysis, and pathogenicity of the fungus, the causal fungus was clearly identified as Rhizopus oryzae Went & Prinsen Geerligs. This is the first report of soft rot by R. oryzae on R. crataegifolius in Korea.

Sphingomonas abietis sp. nov., an Endophytic Bacterium Isolated from Korean Fir

  • Lingmin Jiang;Hanna Choe;Yuxin Peng;Doeun Jeon;Donghyun Cho;Yue Jiang;Ju Huck Lee;Cha Young Kim;Jiyoung Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.10
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    • pp.1292-1298
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    • 2023
  • PAMB 00755T, a bacterial strain, was isolated from Korean fir leaves. The strain exhibits yellow colonies and consists of Gram-negative, non-motile, short rods or ovoid-shaped cells. It displays optimal growth conditions at 20℃, 0% NaCl, and pH 6.0. Results of 16S rRNA gene-based phylogenetic analyses showed that strain PAMB 00755T was most closely related to Sphingomonas chungangi MAH-6T (97.7%) and Sphingomonas polyaromaticivorans B2-7T (97.4%), and ≤96.5% sequence similarity to other members of the genus Sphingomonas. The values of average nucleotide identity (79.9-81.3%), average amino acid identity (73.3-75.9%), and digital DNA-DNA hybridization (73.3-75.9%) were significantly lower than the threshold values for species boundaries; these overall genome-related indexes (OGRI) analyses indicated that the strain represents a novel species. Genomic analysis revealed that the strain has a 4.4-Mbp genome encoding 4,083 functional genes, while the DNA G+C content of the whole genome is 66.1%. The genome of strain PAMB 00755T showed a putative carotenoid biosynthetic cluster responsible for its antioxidant activity. The respiratory quinone was identified as ubiquinone 10 (Q-10), while the major fatty acids in the profile were identified as C18:1ω7c and/or C18:1ω6c (summed feature 8). The major polar lipids of strain PAMB 00755T were diphosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid, and phosphatidylcholine. Based on a comprehensive analysis of genomic, phenotypic, and chemotaxonomic characteristics, we proposed the name Sphingomonas abietis sp. nov. for this novel species, with PAMB 00755T as the type strain (= KCTC 92781T = GDMCC 1.3779T).

Agromyces silvae sp. nov., Rathayibacter soli sp. nov., and Nocardioides terrisoli sp. nov., Isolated from Soil

  • Hyosun Lee;Dhiraj Kumar Chaudhary;Dong-Uk Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.7
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    • pp.1475-1483
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    • 2024
  • Three Gram-stain-positive, aerobic, rod-shaped, and non-motile bacteria, labelled as W11T, SW19T, and YR1T, were isolated from soil, and performed their polyphasic taxonomic investigation. The phylogenetic and 16S rRNA gene sequence analysis showed that strains W11T, SW19T, and YR1T belonged to the genera Agromyces, Rathayibacter, and Nocardioides, respectively. Strain W11T was closely affiliated with Agromyces cavernae SYSU K20354T (98.1%), strain SW19T showed the closest affiliation with Rathayibacter rubneri ZW T2_19T (97.0%), and strain YR1T was most closely related to Nocardioides marmorisolisilvae KIS18-7T (98.0%). The genome sizes of strains W11T, SW19T, and YR1T were 4,181,720 bp, 4,740,677 bp, and 4,228,226 bp, respectively, with DNA G+C contents of 70.5%, 64.2%, and 69.7%, respectively. Average nucleotide identity and digital DNA-DNA hybridization values of W11T, SW19T, and YR1T with their respective reference species were <79.6% and <23.6%, respectively. The predominant cellular fatty acids detected in strain W11T were anteiso-C15:0, iso-C16:0, and anteiso-C17:0. In strain SW19T, they were summed feature 9 (C16:0 10-methyl and/or iso-C17:1ω 9c), anteiso-C17:0, and anteiso-C15:0. Strain YR1T exhibited C18:1ω 9c, C18:0 10-methyl, TBSA, and anteiso-C15:0 as its major cellular fatty acids. Overall, the polyphasic taxonomic comparisons indicated that strains W11T, SW19T, and YR1T represent novel species within the genera Agromyces, Rathayibacter, and Nocardioides, respectively. Accordingly, we propose the names Agromyces silvae sp. nov., with the type strain W11T (=KCTC 49818T =NBRC 115999T), Rathayibacter soli sp. nov., with the type strain SW19T (=KCTC 49860T =NBRC 116108T), and Nocardioides terrisoli sp. nov., with the type strain YR1T (=KCTC 49863T =NBRC 116165T).