• Title/Summary/Keyword: ITS of rDNA

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Antioxidant Properties in Microbial Fermentation Products of Lonicera japonica Thunb. Extract (금은화 추출물을 이용한 미생물 발효 생성물의 항산화 특성)

  • Shin, Jung-Hee;Yoo, Sun-Kyun
    • Journal of the East Asian Society of Dietary Life
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    • v.22 no.1
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    • pp.95-102
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    • 2012
  • The purpose of this study is to investigate antioxidant properties in microbial fermentation products of Lonicera japonica Thunb extract. The bacterium Lactobacillus plantarum NHP1 was isolated from conventional fermented foods. Modern pharmacological studies show that Lonicera japonica Thunb and its active principles of wide pharmacological actions. For instance, they show a strong efficacy in antibacterial, anti-inflammatory, antiviral, anti-endotoxin, blood fat reducing, antipyretic, and antioxidant activities. The extract of Lonicera japonica Thunb was obtained by extracting dried Lonicera japonica Thunb using either hot water or 70% ethanol as a solvent. Fermentation was performed in a 2L fermentor containing 1.2 L of extractat conditions of $30^{\circ}C$ and 100 rpm for 48 hr. The amount of cholorogenic acid was $2.65{\mu}g/g$ in hot water extract. The total phenolic content (GAE, gallic acid equivalent) in hot water and 70% ethanol were $56.5{\pm}4.9$ GAE mg/g and $72.7{\pm}5.3$ GAE mg/g, respectively. After fermentation, the phenolic content increased to 30.2% in hot water and 12.9% in ethanol extract. In the same manner, flavonoid content increased to more than 75% regardless of extract solvent. ABTS (2,2-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid) value noticeably increased to 50% after fermentation.

Detection Rate of Periodontopathogens Associated with Cardiovascular Diseases in Denture. (의치 표면에서 심혈관질환과 관련된 치주질환 원인 세균의 검출)

  • Lim Mi-Young;Kim Hwa-Sook;Jeong Jae-Heon;Yang Ji-Youn;Oh Sang-Ho;Kook Joong-Ki
    • Korean Journal of Microbiology
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    • v.40 no.3
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    • pp.237-243
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    • 2004
  • The aim of this study is to investigate the detection rate of putative periodontopathogens, Porphyromonas gingivalis, Tannerella forsythia, and Actiobacillus actinomycetemcomitans, related to cardiovascular diseases(CVD). Plaques were sampled from 15 subjects (4 sites of denture base and/or tooth) with sterilized explorers and were transported in IX PBS. The detection of periodontopathogens was performed by polymerase chain reaction with species-specific primers based on 16S rDNA. The PCR products were cloned into pGEM-T easy vector and its nucleotides were sequenced in order to confirm the specificity. Our data showed that the detection rate of P. gingivalis and T forsythia in denture base of edentulous patients was 25% and 75%, respectively. And the detection rate of P. gingivalis and T.forsythia in denture base of patient having one more tooth was 91%. The results indicate that plaque of denture base may serve as reservoirs of oral bacteria related to CVD.

Podosphaera pannosa Causes Powdery Mildew and Rusty Spot on Peach Fruits from Korea (복숭아 과실에서 흰가루 증상 및 녹얼룩점 증상을 일으키는 Podosphaera pannosa)

  • Shin, Hyeon-Dong;Cho, Sung-Eun;Choi, In-Young;Seo, Kyoung-Won
    • The Korean Journal of Mycology
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    • v.46 no.2
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    • pp.193-199
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    • 2018
  • The fungus, Podosphaera pannosa, was identified in 1991 as the cause of powdery mildew symptoms on peach (Prunus persica var. persica) fruit from Korea based on the morphological characteristics of the conidial state. Recently, however, in Serbia and France, the cause of 'rusty spot' found on peach fruit was identified as P. leucotricha, and the cause of 'powdery mildew' on nectarine (Prunus persica var. nucipersica) fruit was identified as P. pannosa. To confirm the identity of the Korean pathogen, we collected four samples of powdery mildew from Korean peach fruit: three with the 'powdery mildew' symptom and one with the 'rusty spot' symptom. Morphological examination of the four samples confirmed P. pannosa as the pathogen. Internal transcribed spacer sequences of rDNA were analyzed for molecular characterization. A phylogenetic tree showed that the Korean isolates were clustered into a clade containing P. pannosa from Rosa species, with high sequence similarities of more than 99%. Thus, we showed that the powdery mildew and rusty spot symptoms on peach fruits from Korea are associated with P. pannosa.

Identification and Characterization of Microbial Community in the Coelomic Fluid of Earthworm (Aporrectodea molleri)

  • Yakkou, Lamia;Houida, Sofia;Dominguez, Jorge;Raouane, Mohammed;Amghar, Souad;Harti, Abdellatif El
    • Microbiology and Biotechnology Letters
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    • v.49 no.3
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    • pp.391-402
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    • 2021
  • Earthworms play an important role in soil fertilization, interacting continually with microorganisms. This study aims to demonstrate the existence of beneficial microorganisms living in the earthworm's immune system, the coelomic fluid. To achieve this goal, a molecular identification technique was performed, using cytochrome c oxidase I (COI) barcoding to identify abundant endogenic earthworms inhabiting the temperate zone of Rabat, Morocco. Then, 16S rDNA and ITS sequencing techniques were adopted for bacteria and fungi, respectively. Biochemical analysis, showed the ability of bacteria to produce characteristic enzymes and utilize substrates. Qualitative screening of plant growth-promoting traits, including nitrogen fixation, phosphate and potassium solubilization, and indole acetic acid (IAA) production, was also performed. The result of mitochondrial COI barcoding allowed the identification of the earthworm species Aporrectodea molleri. Phenotypic and genotypic studies of the sixteen isolated bacteria and the two isolated fungi showed that they belong to the Pseudomonas, Aeromonas, Bacillus, Buttiauxella, Enterobacter, Pantoea, and Raoultella, and the Penicillium genera, respectively. Most of the isolated bacteria in the coelomic fluid showed the ability to produce β-glucosidase, β-glucosaminidase, Glutamyl-β-naphthylamidase, and aminopeptidase enzymes, utilizing substrates like aliphatic thiol, sorbitol, and fatty acid ester. Furthermore, three bacteria were able to fix nitrogen, solubilize phosphate and potassium, and produce IAA. This initial study demonstrated that despite the immune property of earthworms' coelomic fluid, it harbors beneficial microorganisms. Thus, the presence of resistant microorganisms in the earthworm's immune system highlights a possible selection process at the coelomic fluid level.

The Isolation and Culture Characterization of a Lipolytic Enzyme Producing Strain from Meju (메주로부터 지질분해 효소 생산 균주의 분리 및 배양학적 특성)

  • Yun, Hye-Ju;Lee, You-Jung;Yeo, Soo-Hwan;Choi, Hye-Sun;Park, Hye-Young;Park, Heui-Dong;Baek, Seong-Yeol
    • Microbiology and Biotechnology Letters
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    • v.40 no.2
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    • pp.98-103
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    • 2012
  • For screening of useful enzymes producing microorganisms from Meju, we isolated high lipase producing strains and their lipolytic enzyme activities were then tested. The lipolytic enzyme activities of isolated microorganisms were therefore tested on the Y124 strain. The gene sequence analysis of ITS from Y124 strain revealed Yarrowia lipolytica. Lipase production by the Y124 strain was studied in media containing various carbon sources. The Y124 strain drastically increased lipolytic enzyme activity in YPO media containing olive oil, as well as in YPDO media containing both olive oil and glucose. Maximal lipase production was achieved in YPD (yeast extract-peptone-D-glucose) media containing 0.7% olive oil when cultured at $30^{\circ}C$ for 8 hrs. The lipase produced from the Y124 strain showed the highest activity in p-NPO (p-nitrophenyl octanoate ($C_8$)), amongst the various p-nitrophenyl esters.

Purification and Characterization of a Fibrinolytic Enzyme Produced by Bacillus amyloliquefaciens HC188 (Bacillus amyloliquefaciens HC188이 생산하는 혈전분해 효소의 정제 및 특성)

  • Shin, So Hee;Hong, Sung Wook;Chung, Kun Sub
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.33-43
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    • 2013
  • A bacterium producing a fibrinolytic enzyme was isolated from Cheonggukjang. The bacterium was identified as a strain of Bacillus amyloliquefaciens by 16S rDNA analysis and designated as B. amyloliquefaciens HC188. The optimum culture medium appeared to be one containing 0.5% (w/v) maltose and 0.5% (w/v) soytone. Bacterial growth in the optimal medium at $37^{\circ}C$ reached the stationary phase after 27 h of incubation and the fibrinolytic enzyme showed optimum activity at 24 h. The enzyme was purified by 20-80% ammonium sulfate precipitation, CM Sepharose fast flow ion exchange chromatography, and Sephacryl S-200HR column chromatography. Its specific activity was 38359.3 units/mg protein and the yield was 5.5% of the total activity of the crude extracts. The molecular weight was 24.7 kDa and the amino acids of the N-terminal sequence were AQSVPYGVSQIKAPA. The fibrinolytic enzyme activity had an optimum temperature of $40^{\circ}C$ and an optimum pH of 8.0, and the enzyme was stable in the ranges $20-40^{\circ}C$ and pH 6.0-8.0. Enzyme activity was increased by $Ca^{2+}$ and $Co^{2+}$ but inhibited by $Cu^{2+}$, EDTA, and PMSF. It is suggested that the purified enzyme is a metallo-serine protease.

A Study on Aronia czarna Bioconversion of Metabolic Compounds by Salted Fish Host Fermenting Bacteria and Its Enhancement During Fermentation (아로니아 기능성 증대를 위한 전통 젓갈 유래 유용발효미생물 활용 생물전환 방안 연구)

  • Lim, Jeong-Muk;Choi, Ui-Lim;Moon, Kwang-Hyun;Kim, Dae-Geun;Ok, Jeong-Kyung;Lee, Jeong-Ho;Oh, Byung-Taek
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.69-69
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    • 2018
  • 아로니아(Aronia czarna)는 anthocyanin, polyphenol, flavonoid, cathechine, chlorogenic acid와 같은 생리활성물질이 풍부하게 존재하며 항산화, 항암, 항균, 피부건강개선, 노화방지 등 다양한 생리활성에 대한 효능이 있는 것으로 알려져 있어 건강 및 기능성식품, 화장품 등의 원료 소재로 각광받고 있다. 생물전환(Bioconversion)은 미생물 또는 효소의 생물학적 촉매 반응을 활용하여 기존 소재의 성분을 변환시키는 기술이다, 최근 생물전환을 활용한 천연소재의 생리활성 물질 기능성, 생체이용률, 안전성을 증대시키기 위한 방안으로 많은 연구가 진행되고 있으며 식품, 의약품, 화장품 등 다양한 분야에서 활성화 되고 있다. 본 연구는 젓갈로부터 분리한 균주를 유전학적 특성을 확인하기 위하여 16S rDNA 염기서열을 분석한 뒤 그중 유산균을 발효공정에 활용하였다. 전북 순창에서 수확된 아로니아 분말과 발효공정을 수행하였으며 아로니아 최적 추출조건 선정, 발효공정 전 후 추출물의 기능성 평가를 진행하기 위하여 DPPH radical scavenging activity, Total polyphenol 함량을 확인하여 항산화 효능 및 유효성분 함량을 평가하였다. 또한 대식세포인 Raw 264.7을 사용하여 MTT assay, Nitric oxide (NO) 생성 억제 효능을 확인하여 세포독성 및 항염증 활성을 평가하였다. 실험결과, 젓갈류 발효물로부터 16종의 다양한 균주를 확보하였으며, 그중 L. rhamnosus, L. plantarum, P. pentosaceus 균을 발효 공정에 활용하여 유용 균주를 선정 결과 P. pentosaceus 종 유산균 처리군에서 무처리군 대비 DPPH radical 소거능 및 polyphenol 함량이 증가됨을 확인하였다. 발효공정 후 항산화 활성은 무처리군 대비 약 119%, polyphenol의 함량은 무처리군 대비 약 119%로 증가됨을 확인되었다. 또한 Raw 264.7 세포실험 결과 발효공정 후 독성활성이 감소되는 경향을 확인되었으며, 항염증 활성이 월등히 증가됨을 확인하였다.

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Isolation and Characterization of an Antagonistic Endophytic Bacterium Bacillus velezensis CB3 the Control of Citrus Green Mold Pathogen Penicillium digitatum (감귤저장병 병원균 Penicillium digitatum 방제를 위한 길항 내생세균 Bacillus velezensis CB3의 분리 및 특성 규명)

  • Lee, Ji-Hyun;Seo, Mun-Won;Kim, Hong-Gi
    • The Korean Journal of Mycology
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    • v.40 no.2
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    • pp.118-123
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    • 2012
  • In order to develop environment friendly fungicide for the control of citrus green mold (Penicillium digitatum) using endophytic bacteria, the 21 bacterial isolates were isolated from citrus leaves in seven different orchards in Jeju Province. Among the 21 bacterial isolates, 5 bacterial isolates presented antifungal activity against green mold pathogen P. digitatum. The CB3 isolate, which showed the most strong antagonistic effect, was selected through opposite culture against the pathogen. The rod-shaped, gram-positive bacterium CB3 was identified as Bacillus velezensis based on morphological, physiological characteristics, 16S rDNA, and gyr A gene sequence analysis. The isolate CB3 showed strong antifungal activity against two citrus postharvest pathogen P. digitatum. Citrus fruits were treated by wound inoculation with P. digitatum pathogen, and the control efficacy of CB3 culture broth was 66.7% ($1{\times}10^8$ cfu/ml). In conclusion, The stability of CB3 and its strong antifungal activity also lead us to believe that it has potential for application as an environment friendly biological control agent.

Sclerotinia Rot on Basil Caused by Sclerotinia sclerotiorum in Korea (Sclerotinia sclerotiorum에 의한 바질 균핵병)

  • Hahm, Soo Sang;Kim, Byoung Ryun;Han, Kwang Seop;Kwon, Mi Kyung;Park, In Hee
    • Research in Plant Disease
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    • v.23 no.1
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    • pp.56-59
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    • 2017
  • During growing season of 2011 to 2013, Sclerotinia rot symptoms consistently have been observed on basil in Yesan-gun, Chungcheongnam-do in Korea. The typical symptom formed initially brownish spot on leaf and stem, and then advancing margins, wilting the whole plant and blighting, eventually died. On the surface of diseased lesions was observed cottony, white, dense mat of mycelial growth, and sclerotia ($30-100{\mu}m$ diameter) formed on stem and leaf. Morphological and cultural characteristic on potato dextrose agar, color of colony was white and colorless chocolate, sclerotium of irregular shape of the oval was black and $5-50{\mu}m$ diameter in size. In pathogenicity test, necrosis and wilt of the inoculated stem were observed in all plants and the pathogen was reisolated from stems. On the basis of mycological characteristics, pathogenicity, and internal transcribed spacer rDNA sequence analysis, this fungus was identified as Sclerotinia sclerotiorum. This is the first report of Sclerotinia rot on basil caused by S. sclerotiorum in Korea.

Induced Systemic Resistance in plants by Bacillus sp. Isolated from Dok-do Islands (독도 자생식물 번행초로부터 분리한 바실러스 속 식물생장촉진근권 세균에 의한 식물병 저항성 유도)

  • Kim, Seung-Kun;Son, Jin-Soo;Kwon, Duck-Kee;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.47 no.4
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    • pp.596-602
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    • 2019
  • In September 2017, the rhizospheric soil of Tetragonia tetragonoides (Pall.) Kuntze was further sampled. One hundred and thirty eight species of microorganisms were isolated from the soil. Indole-3-acetic acid (IAA) production, siderophore production, and phosphate degradation were examined in order to confirm bacterial growth from isolated microorganisms. As a result, most strains were able to produce auxins or siderophores and to solubilize phosphate. In addition, 138 isolated strains were treated with tobacco extract and conferred pathogen resistance to host plants upon treatment. As a result, 35 strains that were able to reduce pathophysiology by more the 60% were selected. Among them, 6 strains with high induced systemic resistance (ISR) activity were found. All of these strains belong to the genus Bacillus according to the 16S rDNA sequence analysis. Bacillus aryabhattai KUDC6619 showed outstanding effects with reduced infection in tobacco and pepper plants. Probably, these Bacillus species play a beneficial role by association with T. tetragonoides for its survival in the harsh conditions found on the island of Dokdo.