• Title/Summary/Keyword: liquid medium addition

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Biochemical and Molecular Characterization of Laccases from Wild Mushrooms

  • Ro, Hyeon-Su
    • 한국균학회소식:학술대회논문집
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    • 한국균학회 2014년도 춘계학술대회 및 임시총회
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    • pp.43-43
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    • 2014
  • White rot fungi have been useful source of enzymes for the degradation of environmental pollutants including polycyclic aromatic hydrocarbons (PAHs) and synthetic dyes. PAHs are widespread organic compounds present in fossil fuels and are routinely generated by incomplete fuel combustion. PAHs are some of the major toxic pollutants of water and soil environments. Synthetic dyes are major water-pollutants, which are toxic to organisms in water environments and interfere photosynthesis of water plants. Removal of PAHs and synthetic dyes has been of interests in the environmental science especially in the environmental microbiology. Mushrooms are fungal groups that function as primary degraders of wood polyphenolic lignin. The ligninolytic enzymes produced by mushroom, including manganese peroxidase, lignin peroxidase, and laccase, mediate the oxidative degradation of lignin. The catalytic power of these enzymes in the degradation of aromatic ring compounds has been sought for the degradation of various organic compounds. In this project, we have screened 60 wild mushroom strains for their degradation activity against two representative PAHs, naphthalene and anthracene, and five aromatic dyes, including alizarin red S, crystal violet, malachite green, methylene blue, rose bengal. The degradation of PAHs was measured by GC while the decolorization of dyes was measured by both UV spectrophotometer and HPLC. As results, 9 wild mushroom strains showed high activity in degradation of PAHs and textile dyes. We also describe the secretive enzyme activities, the transcription levels, and cloning of target genes. In conjunction with this, activities of degradative enzymes, including laccase, lignin peroxidase, and Mn peroxidase, were measured in the liquid medium in the presence of PAHs and dyes. Our results showed that the laccase activity was directed correlated with the degradation, indicating that the main enzyme acts on PAHs and dyes is the laccase. The laccase activity was further simulated by the addition of $Cu^{2+}$ ion. Detailed studies of the enzyme system should be sought for future applications.

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Centipede Scolopendra subspinipes multilans L. KOCH로부터 정제된 항균 물질 Centipedin의 항균 Spectrum 및 작용 Mechanism 연구 (Antibiotic Spectrum and Mechanism of Centipedin)

  • 김기태;홍사원;원호식;김효준;박경배;조기승
    • 미생물학회지
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    • 제34권1_2호
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    • pp.31-36
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    • 1998
  • 지네 Scolopendra subspinipes multilans L. Koch로부터 정제한 Centipedin의 gram-negative, gram-positive bacteria 및 fungi에 대한 광범위 항균 spectrum을 조사하였으며 액체배지를 사용하여 각 균주에 대한 minimal inhibitory concentrations(MICs)값을 측정하였다. 인체의 기관중 주로 폐에 감염되는 gram-negative bacteria인 Klebsiella pneumoniae ATCC 8308 균주에 대한 MIC값이 $2{\mu}g/ml$로서 가장 강한 항균력을 나타내었고 화농균인 Proteus vulgaris NRRL B-123 균주에 대해서도 MIC값이 $16{\mu}g/ml$로 비교적 높은 항균력을 나타내었으며 항균제로서의 작용 mechanism을 시험한 결과 prokaryotic RNA transcription을 억제함을 알 수 있었다. Human blood leukemia(HL-60)와 mouse B lymphocyte myeloma cell 등의 동물세포에 대한 Centipedin의 cytotoxicity는 없는 것으로 확인되었다.

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Biodegradation of Ochratoxin A by Aspergillus tubingensis Isolated from Meju

  • Cho, Sung Min;Jeong, Seong Eun;Lee, Kyu Ri;Sudhani, Hemanth P.K.;Kim, Myunghee;Hong, Sung-Yong;Chung, Soo Hyun
    • Journal of Microbiology and Biotechnology
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    • 제26권10호
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    • pp.1687-1695
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    • 2016
  • Ochratoxin A (OTA), a mycotoxin, contaminates agricultural products and poses a serious threat to public health worldwide. Microbiological methods are known to be a promising approach for OTA biodegradation because physical and chemical methods have practical limitations. In the present study, a total of 130 fungal isolates obtained from 65 traditional Korean meju (a fermented starter for fermentation of soybeans) samples were examined for OTA-biodegradation activity using thin-layer chromatography. Two fungal isolates were selected for OTA-biodegradation activity and were identified as Aspergillus tubingensis M036 and M074 through sequence analysis of the beta-tubulin gene. After culturing both A. tubingensis isolates in Soytone-Czapek medium containing OTA (40 ng/ml), OTA-biodegradation activity was analyzed using high-performance liquid chromatography (HPLC). Both A. tubingensis strains degraded OTA by more than 95.0% after 14 days, and the HPLC analysis showed that the OTA biodegradation by the A. tubingensis strains led to the production of ochratoxin α, which is much less toxic than OTA. Moreover, crude enzymes from the cultures of A. tubingensis M036 and M074 led to OTA biodegradation of 97.5% and 91.3% at pH 5, and 80.3% and 75.3% at pH 7, respectively, in a buffer solution containing OTA (40 ng/ml) after 24 h. In addition, the OTA-biodegrading fungi did not exhibit OTA production activity. Our data suggest that A. tubingensis isolates and their enzymes have the potential for practical application to reduce levels of OTA in food and feed.

Genetic Characterization of Two S-Adenosylmethionine-induced ABC Transporters Reveals Their Roles in Modulations of Secondary Metabolism and Sporulation in Streptomyces coelicolor M145

  • Shin, Su-Kyoung;Park, Hyun-Suh;Kwon, Hyung-Jin;Yoon, Hyun-Jin;Suh, Joo-Won
    • Journal of Microbiology and Biotechnology
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    • 제17권11호
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    • pp.1818-1825
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    • 2007
  • S-Adenosylmethionine (SAM) was previously documented to activate secondary metabolism in a variety of Streptomyces spp. and to promote actinorhodin (ACT) and undecylprodigiosin (RED) in Streptomyces coelicolor. The SAM-induced proteins in S. coelicolor include several ABC transporter components (SCO5260 and SCO5477) including BldKB, the component of a well-known regulatory factor for differentiations. In order to assess the role of these ABC transporter complexes in differentiation of Streptomyces, SCO5260 and SCO5476, the first genes from the cognate complex clusters, were individually inactivated by gene replacement. Inactivation of either SCO5260 or SCO5476 led to impaired sporulation on agar medium, with the more drastic defect in the SCO5260 null mutant (${\Delta}SCO5260$). ${\Delta}SCO5260$ displayed growth retardation and reduced yields of ACT and RED in liquid cultures. In addition, SAM supplementation failed in promoting the production of ACT and RED in ${\Delta}SCO5260$. Inactivation of SCO5476 gave no significant change in growth and production of ACT and RED, but impaired the promoting effect of SAM on ACT production without interfering with the effect on RED production. The present study suggests that SAM induces several ABC transporters to modulate secondary metabolism and morphological development in S. coelicolor.

트리즈 기반 OLED 증착 공정의 글래스 열 변형 개선 (TRIZ-based Improvement of Glass Thermal Deformation in OLED Deposition Process)

  • 이우성;최진영
    • 산업경영시스템학회지
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    • 제40권1호
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    • pp.114-123
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    • 2017
  • The global small and mid-sized display market is changing from thin film transistor-liquid crystal display to organic light emitting diode (OLED). Reflecting these market conditions, the domestic and overseas display panel industry is making great effort to innovate OLED technology and incease productivity. However, current OLED production technology has not been able to satisfy the quality requirement levels by customers, as the market demand for OLED is becoming more and more diversified. In addition, as OLED panel production technology levels to satisfy customers' requirement become higher, product quality problems are persistently generated in OLED deposition process. These problems not only decrease the production yield but also cause a second problem of deteriorating productivity. Based on these observations, in this study, we suggest TRIZ-based improvement of defects caused by glass pixel position deformation, which is one of quality deterioration problems in small and medium OLED deposition process. Specifically, we derive various factors affecting the glass pixel position shift by using cause and effect diagram and identify radical reasons by using XY-matrix. As a result, it is confirmed that glass heat distortion due to the high temperature of the OLED deposition process is the most influential factor in the glass pixel position shift. In order to solve the identified factors, we analyzed the cause and mechanism of glass thermal deformation. We suggest an efficient method to minimize glass thermal deformation by applying the improvement plan of facilities using contradiction matrix in TRIZ. We show that the suggested method can decrease the glass temperature change by about 23% through an experiment.

Process Technology of the Direct Separation and Recovery of Iron and Zinc Metals Contained in High Temperature EAF Exhaust Gas

  • Furukawa, Takeshi;Sasamoto, Hirohiko;Isozaki, Shinichi;Tanno, Fumio
    • 대한전자공학회:학술대회논문집
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    • 대한전자공학회 2001년도 The 6th International Symposium of East Asian Resources Recycling Technology
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    • pp.393-397
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    • 2001
  • The innovatory process, that is the direct separation and recovery of the iron and zinc metals contained in the high temperature exhaust gas generated from the electric arc furnace fer the inn scrap melting and/or the dust treatment, has been proposed. This proposed process consists of the moving coke bed filter that is directly connected to the electric furnace, and the following heavy metal condenser. The exhaust gas passes through the filter and the condenser right after exhausting from the electric furnace. The moving coke bed filter is being controlled at about 1000℃ and collects iron and slag components contained in the high temperature exhaust gas. Heavy metals such as zinc and lead pass through the filter as vapor. Based on the thermodynamic considerations, the iron oxide and the zinc oxide are reduced in the filter. The solution loss reaction rate is comparatively low at about 1000℃ in the coke bed filter by the analysis using the mathematical simulation model. The heavy metal condenser is installed in the position after the coke bed filter, and rapidly cools the gas from about 1000℃ to 450℃ by a full of the cooling medium like the solid ceramic ball in addition to the cooling from the wall. The zinc and lead vapor condense and separate f개m the gas in a liquid state. The investigation of the characteristics of the exhaust gas of the commercial electric arc furnace, the fundamental experiments of the laboratory scale and the bench scale ensured the formation of this proposed process. A small-scale pilot plant examination is carrying out at present to confirm the formation of the process. It is certain that the dust generation of the electric arc furnace is extremely decreased, and it can save the energy consumption of usual dust treatment processes by the realization of this process.

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Synthesis, Characterization and In Vitro Evaluation of Triptolide-lysozyme Conjugate for Renal Targeting Delivery of Triptolide

  • Zheng, Qiang;Gong, Tao;Sun, Xun;Zhang, Zhi-Rong
    • Archives of Pharmacal Research
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    • 제29권12호
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    • pp.1164-1170
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    • 2006
  • A triptolide-lysozyme (TP-LZM) conjugate was synthesized to achieve renal specific delivery and to reduce the side effects of triptolide. Triptolide was coupled to lysozyme through succinic via an ester bond with an average coupling degree of 1 mol triptolide per 1 mol lysozyme. The lysozyme can specifically accumulate in the proximal tubular cells of the kidney, making it a potential carrier for targeting drugs to the kidney. The structure of triptolide succinate (TPS) was confirmed by IR, $^{1}H-NMR$, MS and UV. The concentrations of triptolide in various samples were determined by reversed-phase high-performance liquid chromatography (HPLC). In this study, the physicochemical and stability profiles of TP-LZM under various conditions were investgated the stability and releasing profiles of triptolide-lysozyme (TP-LZM) under various conditions. In vitro release trails showed triptolide-lysozyme was relatively stable in plasma (less than 30% of free triptolide released) and could release triptolide quickly in lysosome (more than 80% of free triptolide released) at $37^{\circ}C$ for 24 h. In addition, the biological activities of the conjugate on normal rat kidney proximal tubular cells (NRK52E) were also tested. The conjugate can effectively reduce NO production in the medium of NRK52E induced by lipopolysaccharide (LPS) but with much lower toxicity. These studies suggest the possibility to promote curative effect and reduce its extra-renal toxicity of triptolide by TP-LZM conjugate.

Comparison of Trichothecene Biosynthetic Gene Expression between Fusarium graminearum and Fusarium asiaticum

  • Lee, Theresa;Lee, Seung-Ho;Shin, Jean Young;Kim, Hee-Kyoung;Yun, Sung-Hwan;Kim, Hwang-Yong;Lee, Soohyung;Ryu, Jae-Gee
    • The Plant Pathology Journal
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    • 제30권1호
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    • pp.33-42
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    • 2014
  • Nivalenol (NIV) and deoxynivalenol (DON) are predominant Fusarium-producing mycotoxins found in grains, which are mainly produced by Fusarium asiaticum and F. graminearum. NIV is found in most of cereals grown in Korea, but the genetic basis for NIV production by F. asiaticum has not been extensively explored. In this study, 12 genes belonging to the trichothecene biosynthetic gene cluster were compared at the transcriptional level between two NIV-producing F. asiaticum and four DON-producing F. graminearum strains. Chemical analysis revealed that time-course toxin production patterns over 14 days did not differ between NIV and DON strains, excluding F. asiaticum R308, which was a low NIV producer. Both quantitative real-time polymerase chain reaction and Northern analysis revealed that the majority of TRI gene transcripts peaked at day 2 in both NIV and DON producers, which is 2 days earlier than trichothecene accumulation in liquid medium. Comparison of the gene expression profiles identified an NIV-specific pattern in two transcription factor-encoding TRI genes (TRI6 and TRI10) and TRI101, which showed two gene expression peaks during both the early and late incubation periods. In addition, the amount of trichothecenes produced by both DON and NIV producers were correlated with the expression levels of TRI genes, regardless of the trichothecene chemotypes. Therefore, the reduced production of NIV by R308 compared to NIV or DON by the other strains may be attributable to the significantly lower expression levels of the TRI genes, which showed early expression patterns.

β-Glucuronidase 저해 활성이 우수한 결명자를 첨가한 상황 균사체 배양액의 생리활성 (Biological Activities of Phellinus linteus Mycelium Culture with Cassiae Semen Extract on β-Glucuronidase Inhibitory Activity)

  • 오은희;박정미;김상희;송인규;한남수;윤향식
    • 한국식품영양학회지
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    • 제25권3호
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    • pp.620-628
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    • 2012
  • We examined the effects of biological activity Phellinus linteus mycelium culture with cassiae semen extract. Firstly, the optimal temperature, initial pH and culture period for mycelial growth in a liquid culture of P. linteus were determined, and they were $30^{\circ}C$, pH 5.0 and 8 days respectively. The five herbal materials were examined against several health functional efficacies, and, as a result, Cassiae semen was chosen, with its superior inhibitory effects in ${\beta}$-glucuronidase inhibitory activity, electron donating activity, ACE inhibitory, and ${\alpha}$-glucosidase inhibitory activities(95.3%, 80.9%, 96.1 and 24.2%, respectively). P. linteus fruit body was investigated on ${\beta}$-glucuronidase inhibitory activity, electron donating activity, ACE inhibitory, and ${\alpha}$-glucosidase inhibitory activities, and they were 54.7%, 81.9%, 30.0% and 20.1%, respectively. Accordingly, C. semen was used in the following experiment, to give an additive functional effect on the P. linteus. As the amount of C. semen in the cultural media increased, mycelial weight and ${\beta}$-glucan contents also increased, but final pH was not influenced. In addition, the ${\beta}$-glucuronidase inhibitory activity, electron donating activity, and ${\alpha}$-glucosidase inhibitory activity increased. P. linteus mycelium culture showed higher activities in the other three tests above, except for electron donating activity, when C. semen was added to the medium before cultivation.

Estrogenic Reduction of Styrene Monomer Degraded by Phanerochaete chrysosporium KFRI 20742

  • Lee Jae-Won;Lee Soo-Min;Hong Eui-Ju;Jeung Eui-Bae;Kang Ha-Young;Kim Myung-Kil;Choi In-Gyu
    • Journal of Microbiology
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    • 제44권2호
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    • pp.177-184
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    • 2006
  • The characteristic biodegradation of monomeric styrene by Phanerochaete chrysosporium KFRI 20742, Trametes versicolor KFRI 20251 and Daldinia concentrica KFRI 40-1 was carried out to examine the resistance, its degradation efficiency and metabolites analysis. The estrogenic reduction effect of styrene by the fungi was also evaluated. The mycelium growth of fungi differentiated depending on the concentration levels of styrene. Additionally P. chrysosporium KFRI 20742 showed superior mycelium growth at less than 200 mg/l, while D. concentrica KFRI 40-1 was more than 200 mg/l. The degradation efficiency reached 99 % during one day of incubation for all the fungi. Both manganese-dependent peroxidase and laccase activities in liquid medium were the highest at the initial stage of incubation, whereas the lowest was after the addition of styrene. However, both activities were gradually recovered after. The major metabolites of styrene by P. chrysosporium KFRI 20742 were 2-phenyl ethanol, benzoic acid, cyclohexadiene-1,4-dione, butanol and succinic acid. From one to seven days of incubating the fungi, the expression of pS2 mRNA widely known as an estrogen response gene was decreased down to the level of baseline after one day. Also, the estrogenic effect of styrene completely disappeared after treatment with supernatant of P. chrysosporium KFRI 20742 from one week of culture down to the levels of vehicle.