It is crucial to develop a miniaturized cultivation method for large and rapid screening of high-yielding mutants of monacolin-K, a powerful anti-hypercholesterolemic secondary metabolite biosynthesized by the fungal cells of Monascus ruber. In order to investigate as many strains as possible in a short time, a miniaturized fermentation method especially suitable for the cultivation of the filamentous Monascus mutants was developed using $50m{\ell}$ culture-tube ($7m{\ell}$ of working volume) instead of the traditional $250m{\ell}$ flask ($50m{\ell}$ of working volume). Generally, in filamentous fungal cell fermentations, morphologies in growth and production cultures should be maintained as thick filamentous and compact-pelleted (usually less than 1 mm in diameter) forms, respectively, for enhanced production of secondary metabolites in final production cultures. In this study, we intended to induce the respective optimal morphologies in the miniaturized culture system for the purpose of rapid screening of overproducers. Miniaturized growth culture system was successfully developed due to the mass production of spores in the statistically optimized solid medium. When large amounts of spores were inoculated into the growth cultures, and brown rice flour (20 g/L) was also supplemented to the growth medium, dense filamentous morphologies were successfully induced in the growth cultures performed with the 50 ml culture tubes. It was implied that the amounts of spores inoculated into the growth tube-cultures and the growth medium components should be the key factors for the induction of the filamentous forms in the growth fermentations. Furthermore, in order to statistically optimize production medium, multiple experiments based on Plackett-Burman design and response surface method (RSM) were carried out, resulting in more than 2 fold enhanced production of monacolin-K in the final production cultures with the optimized production medium. Notably, under the production culture conditions with the statistically optimized medium, optimal pellet sizes below 1 mm in diameter were reproducibly induced, in contrast to the thick and viscous filamentous morphologies observed in the previous production cultures.
Background : The occurrence of lung complications after allogenic bone marrow transplantation(BMT) has been reported as 40-60 percent. The risk factors for lung complications are whole body irradiation, high dose chemotherapy, graft versus host disease, old age and CMV infection. The prevalence of graft versus host disease is less in Korea than in Western countries, but frequency of CMV infection is higher. Therefore, the pattern of lung complications may be different in Korea from those in Western countries. Methods : A retrospective cohort study was performed on one hundred consecutive adult patients who underwent allogenic bone marrow transplantation from December, 1993 to May, 1999 at Asan Medical Center. Lung complications were divided into two groups by the time of development, within 30days (pre-engraftment) and beyond 30 days (post-engraftment), and then subdivided into infectious and non-infectious complication. Infectious complications were defined as having the organism in blood, BAL fluid, pleural fluid or sputum, or compatible clinical findings in patients, which improved with antibiotics or an anti-fungal therapy. Result: 1) Eighty three episodes of lung complications had occurred in 54 patients. 2) Within thirty days after BMT, non-infectious complications were more common than infections, but this pattern was reversed after 30 days. After one year post-BMT, there was no infectious complication except in cases of recurrence of underlying disease or development of chronic GVHD. 3) Among the non-infectious complications, pleural effusion (27 episodes) was most common, followed by pulmonary edema (8 episodes), bronchiolitis obliterans(2 episodes), diffuse alveolar hemorrhage (1 episode) and bronchiloitis obliterans with organizing pneumonia (1 episode). 4) The infectious complications were pneumonia (bacterial: 9 episodes, viral: 4 episodes, fungal : 5 episodes, pneumocystis carinii : 1 episode), pulmonary tuberculosis(3 episodes) and tuberculous pleurisy (3 episodes). 5) Lung complications were more frequent in CMV positive patients and in patients with delayed recovery of neutrophil count. 6) The mortality was higher in the patients with lung complications. Conclusion : Lung complications developed in 54% after allogenic BMT and were associated with higher mortality.
Journal of the Society of Cosmetic Scientists of Korea
/
v.31
no.4
s.54
/
pp.349-357
/
2005
New antioxidative substances for cosmeceuticals were screened from natural resources such as microbial metabolites, mushrooms, and medicinal plants. Four antioxidants were isolated from the fungal metabolite of Eupenicillium shearii and their structures were determined to be new phenolic compounds. The compounds were designated as melanocins A, B, C, and D. Melanocins $A{\sim}D$ exhibited free radical scavenging activity on DPPH and superoxide with $EC_{50}$ values of $21{\sim}94\;and\;7{\sim}84{\mu}M$, respectively, which were stronger activity than those of ${\alpha}-tocopherol$ and BHA. Melanocin A showed anti-wrinkle effects on the UV-irrated hairless mouse skin. A novel hispidin antioxidative compound designated as inoscavin A was isolated from the fruiting body of the mushroom, Inonotus xeranticus. Inoscavin A scavenged superoxide radical with $EC_{50}$ values of $0.03{\mu}g/mL$, and inhibited rat liver microsomal lipid peroxidation with $EC_{50}$ values of $0.3{\mu}g/mL$. Benzastatins $A{\sim}G$, the novel antioxidants isolated from the culture of Streptomyces nitrosporeus showed potent lipid peroxidation inhibitory activity with $EC_{50}$ values of $3{\sim}30{\mu}M$. A cyclopentene compound with strong hypopigmentary effect was isolated from the fungal metabolite of Penicillium sp. and identifed as terrein. Terrein significantly reduced melanin levels in a melanomacyte cell line, Mel-Ab. It showed 10 times stronger activity than kojic acid, but exhibited no cytotoxic effect even in $100{\mu}M$. It was suggested that terrein reduced melanin synthesis by reducing tyrosinase production by MITF down-regulation.
Cheon, Da-Mi;Jang, Da Som;Kim, Hye Young;Choi, Kap Seong;Choi, Sang Ki
Korean Journal of Microbiology
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v.49
no.2
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pp.165-171
/
2013
To isolate a novel antifungal compound, we obtained 571 kinds of endolichenic fungi from Lichen Bioresources Center and examined their antifungal abilities. Four fungi Stereocaulon sp. (1429), Stereocaulon sp. (1430), Cryptosporiopsis sp. (0156), and Graphis sp. (1245) showed high antifungal activity against Candida albicans when they grew in both liquid and solid media. We extracted the culture supernatants of these fungi with chloroform and then ethyl acetate. The chloroform fraction exhibited the highest anti-fungal activities when those fractions were examined for the growth inhibition of Candida albicans with disc diffusion method. To see information for the inhibitor present in chloroform fraction we employed GC-MS for the fractions of Stereocaulon sp. (1429). We found that hexamethylcyclotrisiloxane, decanoic acid, hexadecanonic acid-methyl ester, 14-octadecenoic acid-methyl ester, and octadecenoic acid-methyl ester were present more in chloroform fraction than in ethylacetate fraction. This indicates that those compounds could be possible antifungal candidates since antifungal activity of chloroform extract was two times higher than that of ethyl acetate extract.
Kim, Seon-Hong;Lee, Su-Yeon;Cho, Seong-Min;Hong, Chang-Young;Park, Mi-Jin;Choi, In-Gyu
Journal of the Korean Wood Science and Technology
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v.44
no.1
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pp.113-123
/
2016
This study was to investigate the antifungal activity of A. holophylla essential oil against dermatophytes, such as Epidermophyton floccosum, Trichophyton mentagrophytes and Trichophyton rubrum, and to determine the potential effective compound as dermatitis treatment. To evaluate the potential antifungal activities of A. holophylla essential oil and its fractions, paper disc diffusion and agar dilution method tested with morphological observation. Also, their major constituents were analyzed by GC/MS. To determine synergic effects of active ingredient from A. holophylla essential oil were carried out by checkerboard microtiter plate testing. The morphological changes of the dermatophytes exposed to active fraction G4 were observed by electron microscopes. As the results, the highest activities were identified in the fraction containing ${\alpha}$-bisabolol. A mixture of ${\alpha}$-bisabolol and bornyl acetate showed the synergy effects, expressing high potential effects. Also, morphological observation using electron microscopes showed a dramatic changes of cell membrane of E. floccosum and T. rubrum exposed to fraction G4 containing ${\alpha}$-bisabolol. In conclusion, A. holophylla essential oil and its constituents were expected to be used as antifungal agent or raw material for dermatitis therapy.
Procymidone is a fungicide with anti-androgenic properties widely used to protect fruits from fungal infection, which induces an excessive reactive oxygen species production in male reproductive organs. In this study, to clarify whether procymidone affect the cellular antioxidant system of prostate at onset of puberty, gene expression patterns of the representative antioxidant enzymes such as cytoplasmic glutathione peroxidase (GPx1), phospholipid hydroperoxide GPx (PHGPx), selenoprotein P (SePP), cytoplasmic copper/zinc superoxide dismutase (SOD1), and manganese SOD (SOD2) were investigated in the rat ventral prostates exposed to procymidone using real-time RT-PCR analyses. Seven-week-old Sprague-Dawley rats castrated at 6 weeks old were treated with procymidone (25, 50, or 100 mg/kg per day) orally for 7 consecutive days after testosterone propionate (0.4 mg/kg per day) administration by subcutaneous injection. As compared to normal control animals, GPx1 mRNA expression in prostates significantly increased by the administration with TP and/or procymidone. However, PHGPx and SOD1 mRNA levels significanatly decreased by over 25 mg/kg of procymidone treatment and SePP and SOD2 mRNA levels was significanatly reduced by over 50 mg/kg of procymidone treatment. These findings indicate that procymidone may affect the antioxidant system of prostatic cells in up-regulation mode of GPx1, but in down-regulation modes of PHGPx, SePP, SOD1, and SOD2, suggesting that procymidone may affect differently the cellular antioxidant system of prostate according to the exposure doses.
A soil bacterium, Bacillus subtilis HJ927, exhibiting strong antagonistic property against pathogenic fungi was isolated from pepper fields infested with Phytophthora capsici. Pepper plants inoculated with P. capsici revealed severe root mortality while plants co-inoculated with B. subtilis HJ927 and P. capsici showed drastically reduced root mortality. Low molecular weight substances released by B. subtilis HJ927 mediated the plant protective effect. The anti-fungal compounds released by B. subtilis HJ927 were identified as 3-methylbutyric acid, 2-methylbutyric acid, and methyl 2-hydroxy, 3-phenylpropanoate by high-performance liquid chromatography and gas chromatography-mass spectrometry. In addition to these compounds, B. subtilis HJ927 also produced ${\beta}$-1,3-glucanase, a hydrolytic enzyme implicated in antifungal activity.
Bukholderia pyrrocinia CAB08106-4 has an anti-fungal effect on Garlic White Rot caused by Sclereotium cepivorum and Sclereotium sp. It is environmentally friendly microbial product that prevents and controls a variety of phytopathogens including Garlic White Rot caused by Sclereotium cepivorum and Sclereotium sp. The aim of this study was to assess the environmental toxicity using Cyprinus carpio and Daphnia magna. Bukholderia pyrrocinia CAB08106-4 ($1.0{\times}10^9cfu/mL$) was adminatrated to Cyprinus carpio and Daphnia magna according to the toxicity test guideline for peciticide. $LC_{50}$ of Bukholderia pyrrocinia CAB08106-4 is over $6.67{\times}10^4cfu/mL$ in Cyprinus carpio and Daphnia magna and no adverse effect was observed. Based on these results, we concluded that Bukholderia pyrrocinia CAB08106-4 has no toxiciy for Cyprinus carpio and Daphnia magna.
Kim, Jinse;Park, Jong Woo;Park, Seokho;Choi, Dong Soo;Choi, Seung Ryul;Kim, Yong Hoon;Lee, Soo Jang;Park, Chun Wan;Lee, Jung- Soo;Cho, Byoung-Kwan
Journal of Biosystems Engineering
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v.41
no.2
/
pp.98-107
/
2016
Purpose: The aim of this study was to find an appropriate polymer film, which could reduce the water condensation for pallet-size modified atmosphere packaging (MAP). Methods: Five different types of films were selected from several commercialized films. Prior to the real food storage test, plastic boxes with wetted plastic balls were used to simulate the high humidity conditions of real food storage. The initial MAP condition was 5% oxygen and 95% nitrogen, and the $O_2$ concentration, the relative humidity and water condensation inside the films were checked on a daily basis. The MAP test for tomatoes was conducted by using the most appropriate film from the five films examined in this study. Results: Every film except Mosspack(R) indicated a similar variation in the $O_2$ concentration over the course of time. The relative humidity near the surfaces of all the films except nylon-6 approached saturation conditions over time. For three kinds of films, namely, low-density polyethylene (LDPE) film, anti-fogging oriented polypropylene (AFOPP) film, and Mosspack(R), the inner surfaces of the films were fully covered with dew after a storage period of a day. Conversely, an area of 4.5% was covered with dew in the case of the poly lactic acid (PLA) film, and there was no dew inside the nylon-6 film. The pallet-size MAP test for tomatoes was conducted by using the nylon-6 film and there was no water condensation inside the nylon-6 film over three weeks of storage. Conclusions: During the pallet scale MAP, water condensation could cause severe fungal infection and wetting of the corrugated box. Hence, it was important to minimize water condensation. This study showed that the MAP films with high WVTR such as nylon-6 and PLA could reduce the water condensation inside the pallet scale MAP.
Kim, Keun-Ki;Kim, Yong-Kyun;Son, Hong-Joo;Choi, Young-Whan;Kang, Kyu-Young
Applied Biological Chemistry
/
v.48
no.1
/
pp.34-39
/
2005
There are various crop diseases in green houses that are caused by the cultural environments, especially high temperature and moisture. To solve the forementioned problems, farmers are overusing agricultural chemicals, causing other damages by the chemical residue. In this study, antagonistic bacteria as biological control agents were isolated to produce the environmentally-friendly crops for use in green houses. Eighteen species of antagonistic bacteria were totally isolated from the soil and plants in the Perilla fields, and AK-17 showed the highest activity among the isolates. According to the results of anti-fungal spectrum against several pathogens by AK-17, the antagonism effect of the isolates was remarkable against grey mold rot by Botrytis cinerea, sclerotinia rot by Sclerotnia sclerotiorum, and stem rot by Rhizoctonia solini. To evaluate the biological control effects of the isolates against the major diseases of Perilla, studies were carried out to evaluate the preventive and the curative effects of the diseases throughout the pot experiments. According to the forementioned experiments, the preventive and the curative effects by the isolates against sclerotinia rot were respectively showed as 55% and 92%. For the grey mold rot, those were 40% and 78%, respectively. As to the evaluation of the growth-promoting effect by AK-17, the length and the biomass of the tested plants were increased to 120% and to 164%, respectively. For the leaf numbers and area were respectively increased to 120% and 220%. Furthermore, AK-17 was identified as Burkhoderia sp. according to the results of physiological properties and genetic methods.
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