• Title/Summary/Keyword: Cultured broth

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Enzymatic formation of compound-K from ginsenoside Rb1 by enzyme preparation from cultured mycelia of Armillaria mellea

  • Upadhyaya, Jitendra;Kim, Min-Ji;Kim, Young-Hoi;Ko, Sung-Ryong;Park, Hee-Won;Kim, Myung-Kon
    • Journal of Ginseng Research
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    • v.40 no.2
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    • pp.105-112
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    • 2016
  • Background: Minor saponins or human intestinal bacterial metabolites, such as ginsenosides Rg3, F2, Rh2, and compound K, are more pharmacologically active than major saponins, such as ginsenosides Rb1, Rb2, and Rc. In this work, enzymatic hydrolysis of ginsenoside Rb1 was studied using enzyme preparations from cultured mycelia of mushrooms. Methods: Mycelia of Armillaria mellea, Ganoderma lucidum, Phellinus linteus, Elfvingia applanata, and Pleurotus ostreatus were cultivated in liquid media at $25^{\circ}C$ for 2 wk. Enzyme preparations from cultured mycelia of five mushrooms were obtained by mycelia separation from cultured broth, enzyme extraction, ammonium sulfate (30-80%) precipitation, dialysis, and freeze drying, respectively. The enzyme preparations were used for enzymatic hydrolysis of ginsenoside Rb1. Results: Among the mushrooms used in this study, the enzyme preparation from cultured mycelia of A. mellea (AMMEP) was found to convert ginsenoside Rb1 into compound K with a high yield, while those from G. lucidum, P. linteus, E. applanata, and P. ostreatus produced remarkable amounts of ginsenoside Rd from ginsenoside Rb1. The enzymatic hydrolysis pathway of ginsenoside Rb1 by AMMEP was $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}$ compound K. The optimum reaction conditions for compound K formation from ginsenoside Rb1 were as follows: reaction time 72-96 h, pH 4.0-4.5, and temperature $45-55^{\circ}C$. Conclusion: AMMEP can be used to produce the human intestinal bacterial metabolite, compound K, from ginsenoside Rb1 with a high yield and without food safety issues.

Solid Culture of Phosphate-solubilizing Fungus, Penicillium sp. PS-113 (인산가용화 사상균 Penicillium sp. PS-113의 고체배양)

  • Kang, Sun-Chul;Choi, Myoung-Chul
    • Microbiology and Biotechnology Letters
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    • v.27 no.1
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    • pp.1-7
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    • 1999
  • A fungus, Penicillium sp. PS-113, isolated from soil showed the high phosphate-solubilizing activity in patato dextrose broth-rock phosphate to produce free phosphates to the culture broth with the concentrations of 585 ppm against rock phosphate. In this medium, the optimum temperature and initial pH to solubilize rock phosphate were 30$^{\circ}C$ and pH 7.5, respectively. In order to make the mass production of the conidia from this fungus, we cultured in on various solid-based media like barley, corn, wheat, rice, rice bran, and compost. As a result, the fungus highly produced conidia ranging from 2.1 to $5.1{\times}10_9$ conidia/g${\cdot}$media on these solid media except compost-based medium, which was 0 times less than others. Effects of inoculation of the phosphate solubilizing fungus as a biofertilizer were studied in perlite-based pot cropped with Zea mays Suwon 19. Inoculation of Penicillium sp. PS-113 increased in plant height (1.4 times), plant weight (5.2~8.1 times) and root length (1.1~1.2 times) at 60-day cultivation, compared to Hogland solution either without $NH_4H_2PO_4$ or displace $NH_4H_2PO_4$ to powdered rock phosphate, a phosphorus source for plant growth.

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Antifungal Activity of Bee Venom and Sweet Bee Venom against Clinically Isolated Candida albicans

  • Lee, Seung-Bae
    • Journal of Pharmacopuncture
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    • v.19 no.1
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    • pp.45-50
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    • 2016
  • Objectives: The purpose of this study was to investigate the antifungal effect of bee venom (BV) and sweet bee venom (SBV) against Candida albicans (C. albicans) clinical isolates. Methods: In this study, BV and SBV were examined for antifungal activities against the Korean Collection for Type Cultures (KCTC) strain and 10 clinical isolates of C. albicans. The disk diffusion method was used to measure the antifungal activity and minimum inhibitory concentration (MIC) assays were performed by using a broth microdilution method. Also, a killing curve assay was conducted to investigate the kinetics of the anti-fungal action. Results: BV and SBV showed antifungal activity against 10 clinical isolates of C. albicans that were cultured from blood and the vagina by using disk diffusion method. The MIC values obtained for clinical isolates by using the broth microdilution method varied from $62.5{\mu}g/mL$ to $125{\mu}g/mL$ for BV and from $15.63{\mu}g/mL$ to $62.5{\mu}g/mL$ for SBV. In the killing-curve assay, SBV behaved as amphotericin B, which was used as positive control, did. The antifungal efficacy of SBV was much higher than that of BV. Conclusion: BV and SBV showed antifungal activity against C. albicans clinical strains that were isolated from blood and the vagina. Especially, SBV might be a candidate for a new antifungal agent against C. albicans clinical isolates.

Production of Pigment by Liquid Culture and Monacolin K in Red Mold Rice by Solid State Fermentation of Monascus ruber Strains (Monascus ruber의 액체배양을 통한 색소 생산 및 고체발효를 통한 홍국쌀의 monacolin K 생산 특성)

  • Park, Youn-Je
    • KSBB Journal
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    • v.28 no.6
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    • pp.400-407
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    • 2013
  • The growth characteristics and production of color pigments by Monascus strains were investigated during liquid culture, and production of monacolin K in red mold rice was carried out by solid state fermentation. Four different Monascus ruber strains were cultured in potato dextrose yeast extract broth (PDYB) media at $25^{\circ}C$ for 15 days. The high producing strain for red pigment was not corresponded to the strain for yellow pigment. Production of red pigment was high in the strain causing the fast pH change in culture broth. Production of monacolin K in red mold rice by solid state fermentation was influenced by a combination of wet cell weight and spore density in inoculum by liquid culture. Most strains showed the high production of monacolin K in red mold rice, when submerged fermentation was carried out for 5 days as inoculum for solid state fermentation. These results suggest that submerged fermentation period of inoculum have an effect on the production of monacolin K in red mold rice by solid state fermentation, and monacolin K in red mold rice could be increased by controlling the condition of submerged fermentation for inoculum.

Examination of Metabolites Activating Production of Antibiotic in the Neomycin Producer, S. fradiae (Neomycin 생산균주 S. fradiae의 항생물질 생산을 활성화시키는 성분조사)

  • 김공환;구양모
    • KSBB Journal
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    • v.6 no.1
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    • pp.69-77
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    • 1991
  • When S. fradiae was cultured in S medium, it stavted to produce neomycin in the middle of stationary phase of growth. Antibitoic production is regulated not only by glucose but also by metabolites formed from glucose. A chemically defined minimal salt broth was developen for the study of metabolites activating produition of antibiotic in a neomycin producer. When growth and production or antibiotic in minimal salt broth was examined with a full grown or a vefctativc mycelium, the medium was found not to be good for the growth, but to be good enough for the production of antibiotic with a full grown mycelium. When many carbotlydrates, organic acids, or alcohol were supplmented with instead of glucose in the medium suspcndcn with a full grown mycelium, the amount of antibiotic produced in the medium containing fumaratc was 5 times more than that in the medium with glucose. Further study indicated that the medium is not good also for the growth but good for the production of antibiotic. The antibiotic produced in this medium was identified to be neomycin. The activation of the production of neomycin by fumarate was further confirmed in a complex medium. Fuinarate is suspected to initiate and to activate the biosynthesis of neomycin at the gene level.

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Inhibition of Pathogenic Bacteria by Pediococcus pentosaceus Strain SH-10 Isolated from Hard Clam Meretrix meretrix Sikhae (백합(Meretrix meretrix) 식해에서 분리한 Pediococcus pentosaceus SH-10에 의한 병원성 세균의 억제 기작)

  • Shin, Dong-Min;Kim, Hee-Dai;Koo, Jae-Geun;Park, Kwon-Sam
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.45 no.6
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    • pp.600-605
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    • 2012
  • In this study, we investigated the mechanism of inhibition of pathogenic bacteria by Pediococcus pentosaceus strain SH-10 isolated from hard Clam Meretrix meretrix sikhae. When P. pentosaceus SH-10 was co-cultured in MRS broth with pathogenic bacteria, including Bacillus cereus, Listeria monocytogenes, Salmonella choleraesuis and Staphyloccus aureus, no viable pathogenic cells were detected after 18 h of incubation. However, pediocin or a pediocin-like bacteriocin was not detected in cultures of P. pentosaceus SH-10 by the agar diffusion method. Organic acids were produced in MRS broth in proportion to the incubation time of P. pentosaceus SH-10. These results indicate that P. pentosaceus SH-10 inhibited the growth of pathogenic bacteria by lowering the pH of the growth medium through the production of organic acids, including sodium lactate, sodium acetate, and sodium citrate.

Evaluation of the Selective Enrichment Culture to Recover Clostridium difficile

  • An, Byoungrak;Kim, Heejung;Lee, Kyungwon
    • Korean Journal of Clinical Laboratory Science
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    • v.46 no.4
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    • pp.140-142
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    • 2014
  • To evaluate the recovery rates to increase toxigenic C. difficile, the selective enrichment broth culture methods were compared with commonly used cytotoxin assays and toxigenic culture. First, the enrichment culture, using the selective medium broth for 2 to 5 days, was performed and then, toxigenic C. difficile was confirmed by C. difficile toxin gene-specific PCR after being cultured on C. difficile selective agar. The sensitivity of C. difficile from the enrichment culture (100%) was higher than that of C. difficile selective agar culture (93.8%), while positive predictive values (PPV) were low; 72.7% (16/22) and 88.2% (15/17). PPV of the enrichment culture are not high. Recently, combinations of C. difficile selective agar culture, C. difficile A & B assays, glutamate dehydrogenase, and nucleic acid amplification method are widely used. The enrichment culture was disadvantageous in PPV, turn-around time, and cost. So, what we performed is not considered as a common method of diagnosis of C. difficile-associated diarrhea.

Effect of Lactococcus lactis 1370 on the Formation of Artificial Plaque (Lactococcus lactis 1370가 인공치태 형성에 미치는 영향)

  • Chung, Jin;Yim, Sung-Yee;Oh, Jong-Suk
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.1
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    • pp.77-85
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    • 2000
  • Streptococcus mutans is the most important causative bacteria of dental caries among the oral bacteria. Lactococcus lactis 1370 was isolated from the oral cavity of child. The effect of Lactococcus lactis 1370 on the formation of artificial plaque by Streptococcus mutans was studied. 1. The insoluble substances and bacteria were much more attached on the wall of disposable cuvette in the culture of Streptococcus mutans than in the combined culture of Streptococcus mutans and Lactococcus lactis 1370. 2. The mean weight of produced artificial plaque on the wires in the beaker was 131.7 mg in the culture of Streptococcus mutans only, whereas being reduced to 6.4 mg in the combined culture of Streptococcus mutans and Lactococcus lactis 1370 (p<0.05). The viable cell didn't show the significant difference between them after culturing. 3. When Streptococcus mutans was cultured in the media containing culture supernatant of Lactococcus lactis 1370 cultured in M17 broth containing 0.5% yeast extract and 5% sucrose, the mean weight of produced artificial plaque was 8.0 mg on the wires, whereas being 125.4 mg in the media without culture supernatant of Lactococcus lactis 1370 (p<0.05). The viable cell didn't show the significant difference between them after culturing. 4. When Streptococcus mutans was cultured in the media containing soluble polymer produced by Lactococcus lactis 1370, the mean weight of produced artificial plaque was significantly reduced compared with being cultured in the media without soluble polymer (p<0.05). The viable cell didn't show the significant difference between them after culturing. 5. The soluble polymer produced by Lactococcus lactis 1370 was glucan. 6. The glucan produced by Lactococcus lactis 1370 was water-soluble glucan containing ${\alpha}$-1,6-glucose linkage as the main linkage. These results suggest that the artificial plaque formed by Streptococcus mutans is inhibited by water-soluble glucan produced by Lactococcus lactis 1370.

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Biological Control of Aphid Using Fungal Culture and Culture Filtrates of Beauveria bassiana

  • Kim, Jeong Jun;Jeong, Gayoung;Han, Ji Hee;Lee, Sangyeob
    • Mycobiology
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    • v.41 no.4
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    • pp.221-224
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    • 2013
  • Aphids are one of the most destructive pests in crop production such as pepper, cucumber, and eggplants. The importance of entomopathogenic fungi as alternative pest control agents is increasing. Conidia of entomopathogenic fungi are influenced by environmental conditions, such as temperature and relative humidity, and cause slow and fluctuating mortality. These factors have prevented wider application and use of biocontrol agents. For investigation of means of mitigation of such problems, we conducted bioassays with 47 fungal culture filtrates in order to evaluate the potential of secondary metabolites produced by entomopathogenic fungi for use in aphid control. Among 47 culture filtrates cultured potato dextrose broth, filtrate of Beauveria bassiana Bb08 showed the highest mortality (78%) against green peach aphid three days after treatments. Filtrate of Bb08 cultured in Adamek's medium showed higher toxicity as 100% to third instar nymphs of the aphid compared with seven other filtrates cultured in different broths amended with colloidal chitin or oil. The culture filtrates and fungal cultures from media amended with colloidal chitin or oil had lower control efficacies than filtrates without these additives in three different media. These results indicate that the fungal culture fluid or culture filtrate of B. bassiana Bb08 cultured in Adamek's medium has potential for development as a mycopesticide for aphid control.

THE INFLUENCE OF INSTRUMENTATION IN THE CANAL ON THE EXTENSION OF INFECTION (근관내(根管內) 기계조작(器械操作)이 감염확산(感染擴散)에 미치는 영향(影響))

  • Kim, Yung-Hai
    • Restorative Dentistry and Endodontics
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    • v.10 no.1
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    • pp.177-181
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    • 1984
  • This study was to confirm the influence of intracanal instrumentation whether the pulp infection could be extended to periapical tissue. Fifty three teeth (24 badly decayed and infected, 29 sound teeth) were employed for this experiment and grouped as follows; 1. The specimen taken from the tip of 12 infected pulp in which reamer was inserted to the canal up to apical 1/3 and cultured as long as 48 hours. After 24 hours culture 11 cases were positive and 1 ease was negative but the time of incubation elapsed as 48 hours a negative case turned to positive. 2. Broth immersed paper disc was placed for 1 minute on the tip of 12 infected teeth with a reamer inserted to the apical end and cultured as usual manner in the incubator. At 24 hour culture the growth was significant in 9 cases and after 48 hours total 12 cases were positive. 3. Reamer was inserted to apical 1/3 on 14 sterile pulp canals and specimens obtained from the root tip were cultured for 24 and 48 hours. The results on both group were negative. 4. Similar maner with No.3 except reamer tip was rest exactly at the apex revealed only 2 cases of positive at 48 hour culture. 5. The tip of 24 reamers which reached to apical 1/3 and apex of infected canal were cultured for 24 and 48 hours. At 24 hour culture the growth was evident. 6. The tip of 14 reamers which inserted to apical 1/3 of sterile canal showed negative at 24 hour culture. The 15 cases of the tip which reached to the apex of sterile canal were found negative except 3 positive cases at 48 hour culture.

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