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Isolation of Stenotrophomonas rhizopilae Strain GFC09 with Ginsenoside Converting Activity and Anti-wrinkle Effects of Converted Ginsenosides (사포닌 전환 활성 Stenotrophomonas rhizopilae Strain GFC09 균주의 분리 동정 및 전환 사포닌의 주름 개선 효과)

  • Min, Jin Woo;Kim, Hye-Jin;Joo, Kwang-Sik;Kang, Hee-Cheol
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.41 no.4
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    • pp.375-382
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    • 2015
  • Ginsenosides (ginseng saponin) as the one of important pharmaceutical compounds of ginseng and is responsible for the pharmacological and biological activities. These ginsenoside produces diverse small molecules ginsenoside which have more pharmacological activities including anti-wrinkle, anti-cancer and anti-oxidant effects. In the present study, we isolated bacteria using esculin agar, to produce ${\beta}$-glucosidase, and we focused on the bio-transformation of ginsenoside. Phylogenetic tree analysis was performed by comparing the 16S rRNA sequences; we identified the strain as Stenotrophomonas rhizopilae strain GFC09. In order to determine the optimal conditions for enzyme activity, the crude enzyme was incubated with 1 mM ginsenoside $Rb_1$. Bioconversion of ginsenoside $Rb_1$ were analyzed using TLC and HPLC. The crude enzyme hydrolyzed the ginsenoside $Rb_1$ along the following pathway: LB: $Rb_1{\rightarrow}Rd{\rightarrow}F_2$ into compound K, TSB: $Rb_1{\rightarrow}Rd{\rightarrow}F_2$. The structure of the hydrolyzed metabolites were identified by NMR. The activity screening tests showed that the conversion product induced the production of type I procollagen in a dose-dependent manner. These results suggested that hydrolyzed ginseng product containing the ginsenoside $F_2$ and compound K could be useful as an active ingredient for wrinkle-care cosmetics.

Antifungal and Antioxidative Activities of Yucca smallina Fern

  • Jin, Yu-Lan;Jung, Woo-Jin;Kuk, Ju-Hee;Kim, Jung-Bong;Kim, Kil-Yong;Park, Ro-Dong
    • Journal of Applied Biological Chemistry
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    • v.49 no.4
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    • pp.165-170
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    • 2006
  • The antifungal activity of crude methanolic extract and fractions from Yucca smalliana Fern. leaves, roots and flowers were investigated in vitro against a panel of plant pathogenic fungi. The minimal inhibitory concentration(MIC) was determined by an agar dilution method. Preliminary liquid culture and agar plate assays showed that the growth of Fu sarium oxysporum, Phytophthora capsici, Rhizoctonia solani and Botrytis cinerea were inhibited by Y. smalliana extracts. The extracts from flowers and leaves showed antifungal activity of 64.0% and 34.0% against F. oxysporum, 66.0% and 62.0% against P. capsici, and 27.0% and 41.0% against B. cinerea, respectively. The methanolic extract from Y. smallina leaves in distilled water was fractionated using solvents of increasing polarity: hexane, ethyl acetate and butanol. These fractions had a broad spectrum of antifungal activity, found to reside entirely in the butanol and aqueous fraction. The aqueous fraction showed inhibition rate of 60.0, 67.8, 84.6 and 58.3% against F. oxysporum, R. solani, C. gloeosporioides, and B. cinerea, respectively, and the butganol fracgtion showed 36.0, 46.0, 66.1 and 58.3%, respectively. Phenolics(e.g. flavonoids, steroids and terpenoids) were observed in the thin layer profile of the different fractions. Leave extract showed a prominent antioxidant activity totally scavenging the free radical of DPPH at a concentration of 1 mg/ml.

An Auxin Producing Plant Growth Promoting Rhizobacterium Bacillus subtilis AH18 which has Siderophore-Producing Biocontrol Activity (Auxin과 Siderophore 생산성 다기능 생물방제균 Bacillus subtilis AH18)

  • Jung Hee-Kyoung;Kim Jin-Rak;Woo Sang-Min;Kim Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.34 no.2
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    • pp.94-100
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    • 2006
  • To isolate a bacterium that produces plant growth promoting hormone, a total of 29 bacteria were obtained from the soil in Gyeongsan, Korea. Among these, 14 strains were selected by their positive reaction on Salkowski to produce auxin. All of these were then tested for their property to produce siderophore using CAS (chrome azurol S) blue agar, and one was chosen for its ability to produce both, auxin and siderophore. This strain, denoted, AHl8, showed 1.5 times higher adventitious root induction rates than controls, using mung-beans. The strain also showed efficient biocontrol properties towards Fusarium-wilt of tomatoes in artificial pot assays. The strain was identified as Bacillus subtilis by 16s rDNA comparison and Biolog analyses. Growth and media conditions for Bacillus subtilis AH1 8 to highly produce siderophore were also investigated.

An extensive investigation on gamma ray shielding features of Pd/Ag-based alloys

  • Agar, O.;Sayyed, M.I.;Akman, F.;Tekin, H.O.;Kacal, M.R.
    • Nuclear Engineering and Technology
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    • v.51 no.3
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    • pp.853-859
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    • 2019
  • A comprehensive study of photon interaction features has been made for some alloys containing Pd and Ag content to evaluate its possible use as alternative gamma radiations shielding material. The mass attenuation coefficient (${\mu}/{\rho}$) of the present alloys was measured at various photon energies between 81 keV-1333 keV utilizing HPGe detector. The measured ${\mu}/{\rho}$ values were compared to those of theoretical and computational (MCNPX code) results. The results exhibited that the ${\mu}/{\rho}$ values of the studied alloys are in the same line with results of WinXCOM software and MCNPX code results at all energies. Moreover, Pd75/Ag25 alloy sample has the maximum radiation protection efficiency (about 53% at 81 keV) and lowest half value layer, which shows that Pd75/Ag25 has superior gamma radiation shielding performance among the other compared alloys.

Selection and Antifungal Activity of Antagonistic Bacterium Bacillus subtilis KMU-13 against Cucumber scab, Cladosporium cucumerinum KACC 40576 (검은별무늬병균 Cladosporium cucumerinum KACC 40576에 대한 길항균주 Bacillus subtilis KMU-13의 선발 및 항진균 활성)

  • Park Sung-Min;Lee Jun-Seuk;Park Chi-Duck;Lee Jung-Hun;Jung Hyuck-Jun;Yu Tae-Shick
    • KSBB Journal
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    • v.21 no.1 s.96
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    • pp.42-48
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    • 2006
  • Bacillus subtilis KMU-13 was isolated from the Lillehammer forest soils at Norway and shown a strong antifungal activity on cucumber scab, Cladosporium cucumerinum KACC 40576. B. subtilis KMU-13 produced a maximum level of antifungal substance under incubation aerobically at $30^{\circ}C$, 180 rpm for 48 hours in LB broth containing 0.5% maltose and 0.5% bactopeptone and initial pH adjusted to 6.0. Butanol extract of cultured broth was confirmed inhibitory zone by plate assay and Rf 0.64 value substance by thin layer chromatography (TLC) represented high antifungal activity against C. cucumerinum KACC 40576 and also shown fungal growth inhibitory activity against Botytis cinerea KACC 40573, C. gloeosporioides KACC 40804, D. byoniae KACC 40669, F. oxysporum KACC 40037, F. oxysporum KACC 40052, F. oxysporum f. sp. radicis-lycopersici KACC 40537, F. oxysporum KACC 40902, M. cannonballus KACC 40940, P. cambivora KACC 40160, R. soiani AG-1 KACC 40101, R. solani AG-4 KACC 40142, and S. scleotiorum KACC by agar diffusion method.

Rapid Identification of Diaporthe citri by Gene Sequence Analysis

  • Zar Zar Soe;Yong Ho Shin;Hyun Su Kang;Mi Jin Kim;Yong Chull Jeun
    • Research in Plant Disease
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    • v.29 no.2
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    • pp.130-136
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    • 2023
  • Citrus melanoses caused by Diaporthe citri, has been one of the serious diseases in many citrus orchards of Jeju Island. To protect melanose in citrus farms, a fast and exact diagnosis method is necessary. In this study, diseased leaves and dieback twigs were collected from a total of 49 farms within March to April in 2022. A total of 465 fungal isolates were obtained from a total of 358 isolated plant samples. Among these fungal isolates, 40 representatives of D. citri isolates which were isolated from 22 twigs and 18 leaves on 23 farms were found based on cultural characteristics on potato dextrose agar and conidial morphology. Additionally, the molecular assay was carried out and compared with those by morphological diagnosis. All isolates were identified as D. citri by analyzing the sequences at the internal transcribed spacer (ITS) rDNA region using primers of ITS1/ITS4 or at β-tubulin using primer Btdcitri-F/R. Therefore, based on the present study, where the results of morphological identification of conidial type were consistent with DNA sequence analysis of certain gene, choosing a suitable method for a fast diagnosis of citrus melanose was suggested.

Isolation and Characterization of a Novel Agar-Degrading Marine Bacterium, Gayadomonas joobiniege gen, nov, sp. nov., from the Southern Sea, Korea

  • Chi, Won-Jae;Park, Jae-Seon;Kwak, Min-Jung;Kim, Jihyun F.;Chang, Yong-Keun;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.23 no.11
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    • pp.1509-1518
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    • 2013
  • An agar-degrading bacterium, designated as strain $G7^T$, was isolated from a coastal seawater sample from Gaya Island (Gayado in Korean), Republic of Korea. The isolated strain $G7^T$ is gram-negative, rod shaped, aerobic, non-motile, and non-pigmented. A similarity search based on its 16S rRNA gene sequence revealed that it shares 95.5%, 90.6%, and 90.0% similarity with the 16S rRNA gene sequences of Catenovulum agarivorans $YM01^T$, Algicola sagamiensis, and Bowmanella pacifica W3-$3A^T$, respectively. Phylogenetic analyses demonstrated that strain $G7^T$ formed a distinct monophyletic clade closely related to species of the family Alteromonadaceae in the Alteromonas-like Gammaproteobacteria. The G+C content of strain $G7^T$ was 41.12 mol%. The DNA-DNA hybridization value between strain $G7^T$ and the phylogenetically closest strain $YM01^T$ was 19.63%. The genomes of $G7^T$ and $YM01^T$ had an average ANIb value of 70.00%. The predominant isoprenoid quinone of this particular strain was ubiquinone-8, whereas that of C. agarivorans $YM01^T$ was menaquinone-7. The major fatty acids of strain $G7^T$ were Iso-$C_{15:0}$ (41.47%), Anteiso-$C_{15:0}$ (22.99%), and $C_{16:1}{\omega}7c/iso-C_{15:0}2-OH$ (8.85%), which were quite different from those of $YM01^T$. Comparison of the phenotypic characteristics related to carbon utilization, enzyme production, and susceptibility to antibiotics also demonstrated that strain $G7^T$ is distinct from C. agarivorans $YM01^T$. Based on its phenotypic, chemotaxonomic, and phylogenetic distinctiveness, strain $G7^T$ was considered a novel genus and species in the Gammaproteobacteria, for which the name Gayadomonas joobiniege gen. nov. sp. nov. (ATCC BAA-2321 = $DSM25250^T=KCTC23721^T$) is proposed.

Microbial conversion of major ginsenosides in ginseng total saponins by Platycodon grandiflorum endophytes

  • Cui, Lei;Wu, Song-quan;Zhao, Cheng-ai;Yin, Cheng-ri
    • Journal of Ginseng Research
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    • v.40 no.4
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    • pp.366-374
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    • 2016
  • Background: In this study, we screened and identified an endophyte JG09 having strong biocatalytic activity for ginsenosides from Platycodon grandiflorum, converted ginseng total saponins and ginsenoside monomers, determined the source of minor ginsenosides and the transformation pathways, and calculated the maximum production of minor ginsenosides for the conversion of ginsenoside Rb1 to assess the transformation activity of endophyte JG09. Methods: The transformation of ginseng total saponins and ginsenoside monomers Rb1, Rb2, Rc, Rd, Rg1 into minor ginsenosides F2, C-K and Rh1 using endophyte JG09 isolated by an organizational separation method and Esculin-R2A agar assay, as well as the identification of transformed products via TLC and HPLC, were evaluated. Endophyte JG09 was identified through DNA sequencing and phylogenetic analysis. Results: A total of 32 ${\beta}$-glucosidase-producing endophytes were screened out among the isolated 69 endophytes from P. grandiflorum. An endophyte bacteria JG09 identified as Luteibacter sp. effectively converted protopanaxadiol-type ginsenosides Rb1, Rb2, Rc, Rd into minor ginsenosides F2 and C-K, and converted protopanaxatriol-type ginsenoside Rg1 into minor ginsenoside Rh1. The transformation pathways of major ginsenosides by endophyte JG09 were as follows: $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}C-K$; $Rb2{\rightarrow}C-O{\rightarrow}C-Y{\rightarrow}C-K$; $Rc{\rightarrow}C-Mc1{\rightarrow}C-Mc{\rightarrow}C-K$; $Rg1{\rightarrow}Rh1$. The maximum production rate of ginsenosides F2 and C-K reached 94.53% and 66.34%, respectively. Conclusion: This is the first report about conversion of major ginsenosides into minor ginsenosides by fermentation with P. grandiflorum endophytes. The results of the study indicate endophyte JG09 would be a potential microbial source for obtaining minor ginsenosides.

Isolation of Bacteria with Protease Activity from Cheonggukjang and Purification of Fibrinolytic Enzyme (청국장으로부터 혈전용해 활성이 우수한 균주 분리 및 혈전용해효소정제)

  • Choi, Yeon Hee;Lee, Jun Seung;Bae, So Young;Yang, Keun Jae;Yeom, Kyu Won;Jo, Dong Hyeok;Kang, Ock Hwa;Baik, Hyung Suk
    • Journal of Life Science
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    • v.23 no.2
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    • pp.259-266
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    • 2013
  • To isolate the fibrinolytic enzyme, 268 strains from 21 samples were morphologically isolated from Cheonggukjang collected from Korea and Japan. Among the 268 strains, protease-producing bacteria were isolated in nutrient agar medium including 1% skimmed milk. As a result of this, 22 strains were isolated. Apiweb site was used to identify these strains based on their biochemical properties. In addition, 16S rRNA sequencing was performed to identify the strain. Most of the identified strains were Bacillus subtilis and B. amyloliquefaciens. Fibrinolytic enzyme activity was measured with the fibrin plate method. Five strains were finally selected: A2-14, A2-20, C1-05, C1-09, and F2-01. Of those five strains, the A2-20 strain, which is close to B. amyloliquefaciens, showed the strongest fibrinolytic activity. The fibrinolytic enzyme produced by the A2-20 strain was partially purified from culture supernatant by gel filtration and ion exchange chromatography. The optimal pH and temperature values of the partially purified enzyme were 7.0 and $35^{\circ}C$, respectively. Purified protein analysis was carried out with SDS-PAGE and zymography. A genetic analysis was also conducted by PCR based on the consensus sequence of fibrinolytic enzyme. Corresponding genes with a partial sequence of the A2-20 strain were identified.

Identification and Characterization of Entomopathogenic Fungi Isolated from Bemisia tabaci in Korea (담배가루이(Bemisia tabaci )에서 분리한 곤충병원성진균의 동정 및 병원성 검정)

  • Park, Hyun-Rho;Ryu, Young-Hyun;Yeon, Il-Kyen;Nam, Sung-Hee;Kim, Dong-Geun;Han, Myung-Sae
    • Korean journal of applied entomology
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    • v.53 no.1
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    • pp.27-34
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    • 2014
  • Entomopathogenic fungi were isolated from Bemisia tabaci in an Oriental melon field, and their growth characteristics, factors related to a natural outbreak, and infectivity against Bemisia tabaci, Tetranychus urticae, and Frankliniella intonsa were investigated. The isolates had erect conidiophores bearing whorls of 4-6 phialides with a swollen base where cylindrical conidia of $3.0-3.4{\mu}m$ were attached. The isolates were identified as Isaria fumosorosea on the basis of morphological characteristics and an ITS sequence with 99% similarity. I. fumosorosea IFs-08 grew well on Sabouraud dextrose agar+yeast extract medium(3.2 mm/day/$24^{\circ}C$); it grew better at $35^{\circ}C$ than at $15^{\circ}C$. The isolates of I. fumosorosea-IFs were highly infective and killed 93.9-96.7% B. tabaci, 84.9-92.0% T. urticae, and 81.5-84.4% F. intonsa in bioassay, whereas three isolates (Isaria tenuipes, Isaria farinosa, and Isaria fumosorosea) from KACC showed a low infectivity of 10-20%. To the best of our knowledge, this is the first report of I. fumosorosea isolated from B. tabaci in Korea.