• Journal of Microbiology and Biotechnology
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• v.30 no.4
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• pp.571-582
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• 2020

Quorum sensing (QS)-mediated infections cause severe diseases in human beings. The control of infectious diseases by inhibiting QS using antipathogenic drugs is a promising approach as antibiotics are proving inefficient in treating these diseases. Marine fungal (Pestalotiopsis sydowiana PPR) extract was found to possess effective antipathogenic characteristics. The minimum inhibitory concentration (MIC) of the fungal extract against test pathogen Pseudomonas aeruginosa PAO1 was 1,000 ㎍/ml. Sub-MIC concentrations (250 and 500 ㎍/ml) of fungal extract reduced QS-regulated virulence phenotypes such as the production of pyocyanin, chitinase, protease, elastase, and staphylolytic activity in P. aeruginosa PAO1 by 84.15%, 73.15%, 67.37%, 62.37%, and 33.65%, respectively. Moreover, it also reduced the production of exopolysaccharides (74.99%), rhamnolipids (68.01%), and alginate (54.98%), and inhibited the biofilm formation of the bacteria by 90.54%. In silico analysis revealed that the metabolite of P. sydowiana PPR binds to the bacterial QS receptor proteins (LasR and RhlR) similar to their respective natural signaling molecules. Cyclo(-Leu-Pro) (CLP) and 4-Hydroxyphenylacetamide (4-HPA) were identified as potent bioactive compounds among the metabolites of P. sydowiana PPR using in silico approaches. The MIC values of CLP and 4-HPA against P. aeruginosa PAO1 were determined as 250 and 125 ㎍/ml, respectively. All the antivirulence assays were conducted at sub-MIC concentrations of CLP (125 ㎍/ml) and 4-HPA (62.5 ㎍/ml), which resulted in marked reduction in all the investigated virulence factors. This was further supported by gene expression studies. The findings suggest that the metabolites of P. sydowiana PPR can be employed as promising QS inhibitors that target pathogenic bacteria.

• Journal of Ecology and Environment
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• v.41 no.9
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• pp.271-280
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• 2017

Background: Soil microorganisms play key roles in nutrient cycling and are distributed throughout the soil profile. Currently, there is little information about the characteristics of the microbial communities along the soil depth because most studies focus on microorganisms inhabiting the soil surface. To better understand the functions and composition of microbial communities and the biogeochemical factors that shape them at different soil depths, we analyzed microbial activities and bacterial and fungal community composition in soils up to a 120 cm depth at a fallow field located in central Korea. To examine the vertical difference of microbial activities and community composition, ${\beta}$-1,4-glucosidase, cellobiohydrolase, ${\beta}$-1,4-xylosidase, ${\beta}$-1,4-N-acetylglucosaminidase, and acid phosphatase activities were analyzed and barcoded pyrosequencing of 16S rRNA genes (bacteria) and internal transcribed spacer region (fungi) was conducted. Results: The activity of all the soil enzymes analyzed, along with soil C concentration, declined with soil depth. For example, acid phosphatase activity was $125.9({\pm}5.7({\pm}1SE))$, $30.9({\pm}0.9)$, $15.7({\pm}0.6)$, $6.7({\pm}0.9)$, and $3.3({\pm}0.3)nmol\;g^{-1}\;h^{-1}$ at 0-15, 15-30, 30-60, 60-90, and 90-120 cm soil depths, respectively. Among the bacterial groups, the abundance of Proteobacteria (38.5, 23.2, 23.3, 26.1, and 17.5% at 0-15, 15-30, 30-60, 60-90, and 90-120 cm soil depths, respectively) and Firmicutes (12.8, 11.3, 8.6, 4.3, and 0.4% at 0-15, 15-30, 30-60, 60-90, and 90-120 cm soil depths, respectively) decreased with soil depth. On the other hand, the abundance of Ascomycota (51.2, 48.6, 65.7, 46.1, and 45.7% at 15, 30, 60, 90, and 120 cm depths, respectively), a dominant fungal group at this site, showed no clear trend along the soil profile. Conclusions: Our results show that soil C availability can determine soil enzyme activity at different soil depths and that bacterial communities have a clear trend along the soil depth at this study site. These metagenomics studies, along with other studies on microbial functions, are expected to enhance our understanding on the complexity of soil microbial communities and their relationship with biogeochemical factors.

• Korean Journal of Microbiology
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• v.53 no.3
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• pp.193-199
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• 2017

This study was aimed to evaluate the antimicrobial activity of Penicillium purpurogenum ED76 on several clinically important microorganisms. The endophytic fungus P. purpurogenum ED76 was previously isolated from Swietenia macrophylla leaf. The antimicrobial efficacy of P. purpurogenum ED76 dichloromethane extract was determined via disc diffusion and broth microdilution assay. A kill curve study was conducted and the morphology of extract treated bacterial cells were viewed under scanning electron microscope. The dichloromethane extract showed significant inhibitory activity on 4 test bacteria and 2 test yeasts. The minimal inhibitory concentration of the extract ranged from 125 to $1,000{\mu}g/ml$, which indicates the different susceptibility levels of the test microorganisms to the fungal extract. The kill curve study has revealed a concentration-dependent inhibition for all test microorganisms. With the increase of the extract concentration, the microbial growth was significantly reduced. The scanning electron micrograph of dichloromethane extract-treated Staphylococcus aureus cells showed the total damage of the cells. The cell wall invagination of the bacterial cells also indicates the loss of cellular materials and metabolic activity. The gas chromatography mass spectrometry analysis of the extract also showed that the major compound was stigmasterol, which constitutes 45.30% of the total area. The dichloromethane extract of P. purpurogenum ED76 exhibited significant inhibitory activity on several clinically important bacteria and yeasts. The study proposed a possible mode of action that the extract cause significant damage to the morphology of S. aureus cells.

• Journal of Ecology and Environment
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• v.44 no.2
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• pp.72-80
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• 2020

Background: Antibiotics are widely used to treat animals from infections. After fertilizing, antibacterials can remain in the soil while adversely affecting the soil microorganisms. The concentration of oxytetracycline (OTC) in the soil and its effect on the soil microbial community was assessed. To assess the impact of OTC on the soil microbial community, it was added to the soil at concentrations of 50, 150, and 300 mg kg^-1 and incubated for 35 days. Results: The concentration of OTC added to the soil decreased from 150 to 7.6 mg kg^-1 during 30 days of incubation, as revealed by LC-MS. The deviations from the control values in the level of substrate-induced respiration on the 5th day of the experiment were, on average, 26, 68, and 90%, with OTC concentrations at 50, 150, and 300 mg kg^-1, respectively. In samples with 150 and 300 mg kg^-1 of OTC, the number of bacteria from the 3rd to 14th day was 2-3 orders of magnitude lower than in the control. The addition of OTC did not affect the fungal counts in samples except on the 7th and 14th days for the 150 and 300 mg kg^-1 contaminated samples. Genes tet(M) and tet(X) were found in samples containing 50, 150, and 300 mg kg^-1 OTC, with no significant differences in the number of copies of tet(M) and tet(X) genes from the OTC concentration. Conclusions: Our results showed that even after a decrease in antibiotic availability, its influence on the soil microbial community remains.

• Journal of Food Hygiene and Safety
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• v.26 no.4
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• pp.361-365
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• 2011

To evaluate the indoor air quality of food manufacturing plants, the presence of viable bacteria and fungi was assessed in the indoor air of the facilities at which 9 food items were manufactured. Air samples were collected from the general zone, low clean zone and clean zone of each factory with an air sampler, in combination with plate counts agar using for bacteria, and dichloran-glycerol agar for fungi. The samples were incubated at $25^{\circ}C$ for 4 to 7 days. After culture, the colony forming units (CFU) on each plate were counted and corrected with a positive hole conversion table. The average concentration of bacteria was $2.2{\times}10^3\;CFU/m^3$ in the general zone, $1.2{\times}10^3\;CFU/m^3$ in the low clean zone and $7.3{\times}10^2\;CFU/m^3$ in the clean zone. The average concentration of fungal microbes was $2.5{\times}10^3\;CFU/m^3$ in the general zone, $2.6{\times}10^3\;CFU/m^3$ in the low clean zone, and $2.0{\times}10^2\;CFU/m^3$ in the clean zone. No meaningful differences were detected between the general zone and the low clean zone, but the clean zone had significantly lower concentrations than the other zones. Additionally, the identification of the fungi was performed according to morphological method using a giant culture and slide culture. The fungi were identified as belonging to 18 genera, and the genera Cladosporium(33%), Penicillium(29%) and Aspergillus(26%), predominated. Aspergillus isolates were identified to species level, and A. ochraceus, a mycotoxigenic species, was identified. As part of the effort to control the quality of the indoor air of food manufacturing plants, our results show that continued studies are clearly warranted.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.4
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• pp.471-478
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• 2011

The objectives were to compare the ability of various rumen microbial fractions to reduce nitrate and to assess the effect of nitrate on in vitro fermentation characteristics. Physical and chemical methods were used to differentiate the rumen microbial population into the following fractions: whole rumen fluid (WRF), protozoa (Pr), bacteria (Ba), and fungi (Fu). The three nitrogen substrate treatments were as follows: no supplemental nitrogen source, nitrate or urea, with the latter two being isonitrogenous additions. The results showed that during 24 h incubation, WRF, Pr and Ba fractions had an ability to reduce nitrate, and the rate of nitrate disappearance for the Pr fraction was similar to the WRF fraction, while the Ba fraction needed an adaptation period of 12 h before rapid nitrate disappearance. The WRF fraction had the greatest methane ($CH_4$) production and the Pr fraction had the greatest prevailing $H_2$ concentration (p<0.05). Compared to the urea treatment, nitrate diminished net gas and $CH_4$ production during incubation (p<0.05), and ammonia-N ($NH_3$-N) concentration (p<0.01). Nitrate also increased acetate, decreased propionate and decreased butyrate molar proportions (p<0.05). The Pr fraction had the highest acetate to propionate ratio (p<0.05). The Pr fraction as well as the Ba fraction appears to have an important role in nitrate reduction. Nitrate did not consistently alter total VFA concentration, but it did shift the VFA profile to higher acetate, lower propionate and lower butyrate molar proportions, consistent with less $CH_4$ production by all microbial fractions.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.91-94
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• 2002

Arachidonic acid is a polyunsaturated fatty acid(PUFA) containing twenty carbon atoms with four double bonds. The family of w-6 PUFA, including arachidonic acid as well as r-linoleic acid, was served as intermediates in the formation of several key prostaglandin and leukotrienes. Several fungal strains of the genus Mortierella accumulate high amounts of arachidonic acid. In this study experiments were carried out to optimize the culture conditions for the mass production of fungus Mortierella alpina DSA -12 and lipid production with high proportion of polyunsaturated fatty acids, especially arachidonic acid. The batch culture was carried out in 500 L fermenter containing 50 g/L glucose, 18 g/L corn-steep powder and 100 mg/L MnS04 under $25^{\circ}C$, aeration rate of 0.5 vvm and agitation speed of 200 rpm without pH control. As a result, we could be obtained 22 g/L of cell mass with high contents of lipid 12.1 g/L) and arachidonic acid (5.1 g/L) The intermittent fed-batch culture was performed in the medium containing 20 g/L glucose and 10 g/L corn-steep powder. The final glucose concentration was 170 g/L and pH was maintained at 5.5 ${\sim}$ 6.0 by adding 14% ammonia solution. It was shown relatively high cell concentration (70.5 g/L) with high contents of lipid (45.8 g/L) and arachidonic acid 08.3 g/L). Therefore, when compared to batch cultures, the high concentration of arachidonic acid could be obtained by fed-batch culture using M. alpina DSA -12. These results imply that the fed-batch culture of M. alpina DSA -12 was feasible in industrial purpose and could be employed in the commercial production of arachidonic acid.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.38 no.5 s.118
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• pp.1-8
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• 2006

This study was performed to evaluate extensive electrochemical characteristics of 23 commercially available mediators for laccase. Electrochemical properties, interactions with laccases, and ability to degrade lignin were compared for selected mediators. Among them, NNDS has very similar electrochemical properties in terms of reversibility and redox potential (about 470 mV vs. Ag/AgCl at pH=7) compared to ABTS which is a well-known mediator. Specific activity of purified laccase from Cerrena unicolor was determined by both 2,2'-azino-bis-(3-ethylbenz-thiazoline-6-sulfonic acid) (ABTS) and 1-nitroso-2-naphthol -3,6-disulfonic acid (NNDS). The specific activity of the laccase was 23.2 units/mg with ABTS and 21.2 units/mg with NNDS. The electron exchange rate for NNDS with laccase was very similar to that for ABTS, which meant that NNDS had similar mediating capability to ABTS. Determining methanol concentration after reacting with laccase compared to lignin degradation capabilities of both ARTS and NNDS. ARTS or NNDS alone cannot degrade lignin, but in the presence of laccase enhanced the rate of lignin degradation. ABTS showed better activity in the beginning, and the reaction rate of NNDS with lignin was about a half of that of ABTS at 10 minute, but the final concentration of methanol produced in 1 hour was very similar each other. The reason for similar methanol concentration for both ABTS and NNDS can be interpreted as the initial activity of ABTS was better than that of NNDS, but ABTS would be inhibited laccase activity more during the incubation.

• The Korean Journal of Mycology
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• v.22 no.3
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• pp.260-265
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• 1994

Since around 1980 apple rot caused by Botryosphaeria dothidea has become prevalent throughout the growing areas in Korea, during which period chemical controls have been executed with no notable improvement. Results of investigations on resistance of the causal fungus to its control chemical are as followings; The susceptible fungal isolates showed no mycelial growth at $150\;{\mu}g/ml$ of benomyl whereas the resistant isolates showed 7-13 mm growth at $300\;{\mu}g/ml$ and 6-8 mm at $2,400\;{\mu}g/ml$ of this fungicide. At the latter high concentration, spore germination of the resistant isolates was 5-9% while that of the susceptible isolates was 0%. Within the range of $20-2,400\;{\mu}g/ml$ tested, the susceptible isolates were unable to form pycnidia, but the resistant isolates formed abundant pycnidia at the lower concentration with decreasing pycnidia along with the higher concentration.

• The Korean Journal of Pharmacology
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• v.9 no.2
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• pp.39-45
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• 1973

The majority of drugs used in the treatment of superficial fungal infections has limited values due to its low efficacy or development of resistance. For the purpose of searching efficacious agent on the superficial fungal infections induced by dermatophytes which is regarded as the most malicious one, authors examined whether Chinae Rhizoma Extracts have significant on it. Extracts from Smilax china Linne used for the study are water extract (CRWE), ethanol extract (CREE) and methanol extract (CRME). In in vitro studies, the spores of the dermatophytes were inoculated on Sabouraud's glucose agar media which contained three extracts of Chinae Rhizoma in each concentration of $500\;{\mu}g/ml$, $1,000\;{\mu}g/ml$ and $5,000\;{\mu}g/ml$ respectively, and also $1,000\;{\mu}g/ml$ of salicylic acid and undecylenic acid $1,000\;{\mu}g/ml$ as comparable drugs. The growth of the dermatophytes were observed for 3 weeks. The species of the dermatophytes used in this experiment were Epidermophyton floccosum, Microsporum canis, Microsporum cookei, Microsporum gypseum, Microsporum nanum, Trichophyton mentagrophytes, Trichophyton rubrum, Trichophyton tonsurans and Trichophyton verrucosum distributed from The Institute of Tropical Medicine in Belgium. The results of the studies were as follows: 1. The growth of M. canis, M. nanum, T. mentagrophytes, T. rubrum & T. tonsurans were slightly inhibited in CRWE $1,000\;{\mu}g/ml$ and CRWE $5,000\;{\mu}g/ml$, and only slight inhibition on the growth of E. floccosum, M. canis and M. gypseum were observed in CRWE $5,000\;{\mu}g/ml$. 2. Complete inhibition of T. rubrum, moderate inhibition of M. nanum & T. tonsurans, and slight inhibition of E. floccosunl, M. canis, M. cookei & T. mentagrophytes in growth were observed in concentration of CREE $500\;{\mu}g/ml$. The growth of M. gypseum was slightly inhibited, moderate inhibition on the growth of M. canis, M. cookei & T. mentagrophytes, and complete inhibition of E. floccosum, M. nanum, T. rubrum & T. tonsurans in growth were observeed by CREE $1,000\;{\mu}g/ml$. With $5,000\;{\mu}g/ml$ of CREE, the growth of E. floccosum, M. canis, M. cookei, M. gypseum, T. mentagrophytes, T. rubrum & T. tonsurans were completely inhibited except T. verrucosum being showed slight inhibition. 3. In CRME $500\;{\mu}g/ml$, slight inhibition of T. verrucosum, moderate inhibition of M. gypseum and complete inhibition of E. floccosum, M. canis, M. cookei, T. mentagrophytes, T. rubrum & T. tonsurans in growth were observed. The growth of E. floccosum, M. canis, M. cookei, M. gypseum, M. nanum, T. mentagrophytes, T. rubrum & T. tonsurans were completely inhibited except T. verrucosum being showed moderate inhibition in both CRME $1,000\;{\mu}g/ml$ and CRME $5,000\;{\mu}g/ml$. 4. In $1,000\;{\mu}g/ml$ of undecylenic acid, slight inhibition of T. verrucosum and complete inhibition of E, floccosum, M. canis, M. cookei, M. gypseum, M. nanum, T. mentagrophytes, T. rubrum & T. tonsurans in growth were observed. From the above results, it was found that Chinas Rhizoma Alcoholic Extracts(CREE & CRME) exerted significant antifungal activity, and their effects were probably derived from the pharmacological actions of triterpenoidal saponin and steroidal saponin.