• Title/Summary/Keyword: E-Isolation

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Studies on the Amylase Production by Bacteria (세균(細菌)에 의(依)한 Amylase생산(生産)에 관한 연구(硏究))

  • Park, Yoon-Joong
    • Applied Biological Chemistry
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    • v.13 no.2
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    • pp.153-170
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    • 1970
  • 1. Isolation and identification of amylase-producing bacteria. The powerful strain A-12 and S-8 were respectively isolated from air and soil after screening a large number of amylase-producing bacteria. Their bacterial characteristics have been investigated and it has been found that all characteristics of strain A-12 and S-8 are similar to Bac. subtilis of Bergey's manual except for the acid formation from a few carbohydrates and the citrate utilization, i.e., the strain A-12 shows negative in the citrate utilization, and the acid formation from arabinose and xylose, S-8 shows negative in the acid formation from xylose. 2. Amylase production by Liquid cultures with solid materials. Several conditions for amylase production by strain A-12 in stationary cultures have been studied. The results obtained are as follows. (1) The optimum conditions are:temperature $35^{\circ}C$, initial pH 6.5 to 7.0 and incubation time 3 to 4 days. (2) The amylase production is not affected by the preservation period of the stock cultures. (3) Among the various solid material, the defatted soy bean is found to be the best for t1e amylase production. However, the alkali treatment of the defatted soy bean gives no effect contrary to the cage of defatted rape seed. The addition of soluble starch to the alkali extract of defatted soy bean shows the increased amylase production. (4) Up to 1% addition of ethanol to carbon dificient media gives the improved amylase production, whereas the above effect is not found in the case of carbon rich media. (5) The amylase production can be increased 2.5 times when 10% of defatted soy bean is admixed to cheaply available wheat bran. (6) The excellent effect is found for amylase production when 20% of wheat bran is admixed to defatted dry milk which is a poor medium. The activity is found to be $D^{40^{\circ}}_{30'}$ 7,000(L.S.V. 1,800) in 10% medium. (7) No significant effect is observed due to the addition of various inorganic salts. 3. Amylase production by solid cultures. Several conditions for amylase production by strain A-12 in wheat bran cultures have been studied and the results obtained are as follows. (1) The optimum conditions: are temperature $33^{\circ}C$, incubation lime 2 days, water content added 150 to 175% and the thickness of the medium 1.5cm, The activity is found to be $D^{40^{\circ}}_{30'}$ 36,000(L.S.V. 15,000) (2) No significant effect is found in the case of the additions of various organic and inorganic substances.

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Antibacterial and Antioxidative Activities of Quercus acutissima Carruth Leaf Extracts and Isolation of Active Ingredients (상수리나무 잎 추출물의 항균 및 항산화 활성과 활성 물질 분리)

  • Park, Soo-Nam;Kim, So-I;Ahn, You-Jin;Kim, Eun-Hee
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.35 no.2
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    • pp.159-169
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    • 2009
  • In this study, the antibacterial activity, antioxidative effects, inhibitory effects on tyrosinase, inhibitory effects on elastase, and components of Quercus acutissima Carruth leaf extracts were investigated. MIC values of ethyl acetate fraction from Q. acutissima Carruth leaf on P. acnes, S. aureus, P. ovale, and E. coli were 0.13 %, 0.25 %, 0.13 % and 0.25 %, respectively. The results showed that the antibacterial activity of the ethyl acetate fraction was the highest in the S. aureus, P. acnes, and P. ovale. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$) of extract/fractions of Q. acutissima Carruth. leaf was in the order: 50 % ethanol extract (12.13 ${\mu}g/mL$) < ethyl acetate fraction (7.07 ${\mu}g/mL$) < deglycosylated flavonoid aglycone fraction (6.20 ${\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of some Q. acutissima Carruth leaf extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activity was 50 % ethanol extract ($OSC_{50}$, 1.81 ${\mu}g/mL$) < ethyl acetate fraction (1.70 ${\mu}g/mL$) < deglycosylated flavonoid aglycone fraction (0.70 ${\mu}g/mL$). Deglycosylated flavonoid aglycone fraction showed the most prominent scavenging activity. The protective effects of extract/fractions of Q. acutissima Carruth leaf on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Q. acutissima Carruth leaf extracts suppressed photohemolysis in a concentration dependent manner, particularly deglycosylated flavonoid aglycone fraction exhibited the most prominent celluar protective effect (${\tau}50$, 220.00 min at 25 ${\mu}g/mL$). Aglycone fractions obtained from the deglycosylation reaction of ethyl acetate fraction among the Q. acutissima Carruth leaf extracts, showed 3 bands (QA 1, QA2 and QA3) on TLC. TLC chromatogram of ethyl acetate fraction of Q. Carruth. leaf extract revealed 4 bands (QA 1 ${\sim}$ QA 4), Among them, kaempferol (QA 1), quercetin (QA 2), and gallic acid (QA 3) were identified. The inhibitory effect ($IC_{50}$) of aglycone fraction on tyrosinase was 65.7 ${\mu}g/mL$. The inhibitory effect ($IC_{50}$) of aglycone fraction on elastase was 24.50 ${\mu}g/mL$. These results indicate that extract/fractions of Q. acutissima Carruth. can functionized as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. Extract/fractions of Q. acutissima Corruth can be applicable to new functional cosmetics for antioxidant, antiaging, antibacterial activity.

The Effects of Self-regulatory Resources and Construal Levels on the Choices of Zero-cost Products (자아조절자원 및 해석수준이 공짜대안 선택에 미치는 영향)

  • Lee, Jinyong;Im, Seoung Ah
    • Asia Marketing Journal
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    • v.13 no.4
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    • pp.55-76
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    • 2012
  • Most people prefer to choose zero-cost products they may get without paying any money. The 'zero-cost effect' can be explained with a 'zero-cost model' where consumers attach special values to zero-cost products in a different way from general economic models (Shampanier, Mazar and Ariely 2007). If 2 different products at the regular prices of ₩200 and ₩400 simultaneously offer ₩200 discounts, the prices will be changed to ₩0 and ₩200, respectively. In spite of the same price gap of the two products after the ₩200 discounts, people are much more likely to select the free alternative than the same product at the price of ₩200. Although prior studies have focused on the 'zero-cost effect' in isolation of other factors, this study investigates the moderating effects of a self-regulatory resource and a construal level on the selection of free products. Self-regulatory resources induce people to control or regulate their behavior. However, since self-regulatory resources are limited, they are to be easily depleted when exerted (Muraven, Tice, and Baumeister 1998). Without the resources, consumers tend to become less sensitive to price changes and to spend money more extravagantly (Vohs and Faber 2007). Under this condition, they are also likely to invest less effort on their information processing and to make more intuitive decisions (Pocheptsova, Amir, Dhar, and Baumeister 2009). Therefore, context effects such as price changes and zero cost effects are less likely in the circumstances of resource depletion. In addition, construal levels have profound effects on the ways of information processing (Trope and Liberman 2003, 2010). In a high construal level, people tend to attune their minds to core features and desirability aspects, whereas, in a low construal level, they are more likely to process information based on secondary features and feasibility aspects (Khan, Zhu, and Kalra 2010). A perceived value of a product is more related to desirability whereas a zero cost or a price level is more associated with feasibility. Thus, context effects or reliance on feasibility (for instance, the zero cost effect) will be diminished in a high level construal while those effects may remain in a low level construal. When people make decisions, these 2 factors can influence the magnitude of the 'zero-cost effect'. This study ran two experiments to investigate the effects of self-regulatory resources and construal levels on the selection of a free product. Kisses and Ferrero-Rocher, which were adopted in the prior study (Shampanier et al. 2007) were also used as alternatives in Experiments 1 and 2. We designed Experiment 1 in order to test whether self-regulatory resource depletion will moderate the zero-cost effect. The level of self-regulatory resources was manipulated with two different tasks, a Sudoku task in the depletion condition and a task of drawing diagrams in the non-depletion condition. Upon completion of the manipulation task, subjects were randomly assigned to one of a decision set with a zero-cost option (i.e., Kisses ₩0, and Ferrero-Rocher ₩200) or a set without a zero-cost option (i.e., Kisses ₩200, and Ferrero-Rocher ₩400). A pair of alternatives in the two decision sets have the same price gap of ₩200 between a low-priced Kisses and a high-priced Ferrero-Rocher. Subjects in the no-depletion condition selected Kisses more often (71.88%) over Ferrero-Rocher when Kisses was free than when it was priced at ₩200 (34.88%). However, the zero-cost effect disappeared when people do not have self-regulatory resources. Experiment 2 was conducted to investigate whether constual levels influence the magnitude of the 'zero-cost effect'. To manipulate construal levels, 4 different 'why (in the high construal level condition)' or 'how (in the low construal level condition)' questions about health management were asked. They were presented with 4 boxes connected with downward arrows. In a box at the top, there was one question, 'Why do I maintain good physical health?' or 'How do I maintain good physical health?' Subjects inserted a response to the question of why or how they would maintain good physical health. Similar tasks were repeated for the 2nd, 3rd, and 4th responses. After the manipulation task, subjects were randomly assigned either to a decision set with a zero-cost option, or to a set without it, as in Experiment 1. When a low construal level is primed with 'how', subjects chose free Kisses (60.66%) more often over Ferrero-Rocher than they chose ₩200 Kisses (42.19%) over ₩400 FerreroRocher. On contrast, the zero-cost effect could not be observed any longer when a high construal level is primed with 'why'.

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Hydrological Significance on Interannual Variability of Cations, Anions, and Conductivity in a Large Reservoir Ecosystem (대형 인공호에서 양이온, 음이온 및 전기전도도의 연변화에 대한 수리수문학적 중요성)

  • An, Kwang-Guk
    • Korean Journal of Ecology and Environment
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    • v.34 no.1 s.93
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    • pp.1-8
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    • 2001
  • During April 1993 to November 1994, cations, anions, and conductivity were analyzed to examine how summer monsoon influences the ionic content of Taechung Reservoir, Korea. Interannual variability of ionic content reflected hydrological characteristics between the two years(high-flood year in 1993 vs. draught year in 1994). Cations, anions and conductivity were lowest during peak inflow in 1993 and highest during a drought in 1994. Floods in 1993 markedly decreased total salinity as a result of reduced Ca$^{2+}$ and HCO$_{3}\;^{-}$ and produced extreme spatial heterogeneity (i.e., longitudinal, vertical, and horizontal variation) in ionic concentrations. The dominant process modifying the longitudinal (the headwaters-to-downlake) and vertical (top-to-bottom) patterns in salinity was an interflow current during the 1993 monsoon. The interflow water plunged near a 27${\sim}$37 km-location (from the dam) of the mid-lake and passed through the 10${\sim}$30m stratum of the reservoir, resulting in an isolation of epilimnetic high conductivity water (>100 ${\mu}$S/cm) from advected river water with low conductivity (65${\sim}$75 ${\mu}$S/cm), During postmonsoon 1993, the factors regulating salinity differed spatially; salinity of downlake markedly declined as a result of dilution through the mixing of lake water with river water, whereas in the headwaters it increased due to enhanced CaCO$_{3}$ (originated from limestone/metamorphic rock) of groundwaters entering the reservoir. This result suggests an importance of the basin geology on ion compositions with hydrological characteristics. In 1994, salinity was markedly greater (p<0.001) relative to 1993 and ionic dilution did not occur during the monsoon due to reduced inflow. Overall data suggest that the primary factor influencing seasonal ionic concentrations and compositions in this system is the dilution process depending on the intensity of monsoon rainfall.

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소설과 말기 암환자를 통해 본 한국인의 죽음의 의미

  • Jeon, Hye-Won;Kim, Bun-Han
    • Korean Journal of Hospice Care
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    • v.3 no.2
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    • pp.34-54
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    • 2003
  • Every one experiences death one day, however no one can knows exactly what it is because people can not experience death until it comes, it is therefore impossible to judge correctly on the phenomenon of the death. On the whole, man experiences indirect death through the mass communications such as TV drama, fiction, magazine etc because those methods can easily access by every one. In addition to this, people usually acquire the negative awareness of death through the dramatic change of story like dying of cancer for dramatic effect by giving scare and fear to the cancers. The purpose of this study is to provide basic information on the spiritual care that enables the facing death patients to accept death as a part of life and divert hope from scare about after death by comparing and analyzing of two aspects of death meaning I.e, Korean fiction and the end stage cancer patients. Additionally, for medical staff to understand the facing death cancer patients by making to aware patients correctly and provide the better quality of care. The study was performed from September 28, 2002 to February, 28 2003. The materials of this study were collected by direct data obtained from observation, interviews, note and diary of end stage of cancer patients and written materials acquired from Korean contemporary fiction. Participants of this study were 4 end stage cancer patients including 2 lung cancer patients, 1 liver cancer patient and 1 esophagus cancer patient. The methodology used in this study was divided into two types; Huberman & Miles methodology was used for fiction to find and categorize subject, and Colaizzi, one of phenomenological methodology was used for end stage cancer patients to find the major meaning, subject and categorization. 1.The death investigated in the fiction, was found as a progress of negative emotion, acceptance and sublimation, life related subjects in the negative emotion were tenacity for life, anxiety, lingering attachment, responsibility, abandonment and death related subjects were shock, isolation, fear, scare and rejection. Acceptance related subjects were acceptance, destiny, secularism, preparation and arrangement, and sublimation related subjects were sublimation through Christian and Buddhism. 2.The death showed in the participants was negative emotion, acceptance and sublimation, life related subjects were repentance, anxiety, responsibility and hopelessness, and death related subjects were dejection, solitude, anger, fear and scare. The acceptance was a type of religious acceptance that admitted instantly by reaching an understanding with the God, and death was accepted as a progress of preparation, arrangement, acceptance and hope. Sublimation related subjects were Christian sublimation and relief or destiny incurred from self-reflective sublimation through communications and thoughts. 3.The death in view of fiction and participants were positively accepted both death and negative emotion, and the study disclosed the fact that death was sublimated dependent on religion. 4.The progress of negative emotion, acceptance and sublimation was disclosed more complicated and various in the real end stage cancer patients and acceptance only found in the patients on the form of religious acceptance, according to the results compared with fiction and real end stage cancer patients. The death showed in the fiction was standardized, gradated and similar progress with psychological status of Kubler-Ross. However, death in the participants was showed complex and various feelings simultaneously, and sometimes they accepted death positively. The sublimation through religion was found in Buddhism and Christian in the fiction and mostly Christian in the participants due to a number of Hospice patients. It was found that negative emotion various types of death was more found in the participants than fiction. It is therefore necessary to study on the response of death in various types. In the participants death was incurred more systematic and variously, we knew that nursing practice focused on experience of participants is required and reality on death is much profound than we analyzed and presented, lots of situations and reactions should be premised because we can not completely rule out the negligence possibility of care mediation of participants. In caring for the facing death patients, we discovered and confirmed again through this study that the spiritual care should be needed as a mediation method.

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Characterization of SID2 that is required for the production of salicylic acid by using β-GLUCURONIDASE and LUCIFERASE reporter system in Arabidoposis (리포트 시스템을 이용한 살리실산 생합성 유전자 SID2의 발현 해석)

  • Hong, Mi-Ju;Cheong, Mi-Sun;Lee, Ji-Young;Kim, Hun;Jeong, Jae-Cheol;Shen, Mingzhe;Ali, Zahir;Park, Bo-Kyung;Choi, Won-Kyun;Yun, Dae-Jin
    • Journal of Plant Biotechnology
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    • v.35 no.3
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    • pp.169-176
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    • 2008
  • Salicylic acid(SA) is a phytohormone that is related to plant defense mechanism. The SA accumulation is triggered by abiotic and biotic stresses. SA acts as a signal molecular compound mediating systemic acquired resistance and hypersensitive response in plant. Although the role of SA has been studied extensively, an understanding of the SA regulatory mechanism is still lacking in plants. In order to comprehend SA regulatory mechanism, we have been transformed with a SID2 promoter:GUS::LUC fusion construct into siz1-2 mutant and wild plant(Col-0). SIZ1 encodes SUMO E3 ligase and negatively regulates SA accumulation in plants. SID2(SALICYLIC ACID INDUCTION DEFICIENT2) is a crucial enzyme of SA biosynthesis. The Arabidopsis SID2 gene encodes isochorismate synthase(ICS) that controls SA level by conversion of chorismate to isochorismate. We compared the regulation of SID2 in wild-type and siz1-2 transgenic plants that express SID2 promoter:GUS::LUC constructs respectively. The expressions of $\beta$-GLUCURONIDASE and LUCIFERASE were higher in siz 1-2 transgenic plant without any stress treatment. SID2 promoter:GUS::LUC/siz1-2 transgenic plant will be used as a starting material for isolation of siz1-2 suppressor mutants and genes involved in SA-mediated stress signaling pathway.

EVALUATION OF OSTEOGENIC ACTIVITY AND MINERALIZATION OF CULTURED HUMAN PERIOSTEAL-DERIVED CELLS (배양된 인간 골막기원세포의 조골활성 및 골기질 형성의 평가)

  • Park, Bong-Wook;Byun, June-Ho;Lee, Sung-Gyoon;Hah, Young-Sool;Kim, Deok-Ryong;Cho, Yeong-Cheol;Sung, Iel-Yong;Kim, Jong-Ryoul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.6
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    • pp.511-519
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    • 2006
  • Autogenous bone grafts have been considered the gold standard for maxillofacial bony defects. However, this procedure could entail a complicated surgical procedure as well as potential donor site morbidity. Possibly the best solution for bone-defect regeneration is a tissue engineering approach, i.e. the use of a combination of a suitable scaffold with osteogenic cells. A major source of osteogenic cells is the bone marrow. Bone marrow-derived mesenchymal stem cells are multipotent and have the ability to differentiate into osteoblastic, chondrocytic, and adipocytic lineage cells. However, the isolation of cells from bone marrow has someproblems when used in clinical setting. Bone marrow aspiration is sometimes potentially more invasive and painful procedure and carries of a risk of morbidity and infection. A minimally invasive, easily accessible alternative would be cells derived from periosteum. The periosteum also contains multipotent cells that have the potential to differentiate into osteoblasts and chondrocytes. In the present study, we evaluated the osteogenic activity and mineralization of cultured human periosteal-derived cells. Periosteal explants were harvested from mandibule during surgical extraction of lower impacted third molar. The periosteal cells were cultured in the osteogenic inductive medium consisting of DMEM supplemented with 10% fetal calf serum, 50g/ml L-ascorbic acid 2-phosphate, 10 nmol dexamethasone and 10 mM -glycerophosphate for 42 days. Periosteal-derived cells showed positive alkaline phosphatase (ALP) staining during 42 days of culture period. The formation of ALP stain showed its maximal manifestation at day 14 of culture period, then decreased in intensity during the culture period. ALP mRNA expression increased up to day 14 with a decrease thereafter. Osteocalcin mRNA expression appeared at day 7 in culture, after that its expression continuously increased in a time-dependent manner up to the entire duration of culture. Von Kossa-positive mineralization nodules were first present at day 14 in culture followed by an increased number of positive nodules during the entire duration of the culture period. In conclusion, our study showed that cultured human periosteal-derived cells differentiated into active osteoblastic cells that were involved in synthesis of bone matrix and the subsequent mineralization of the matrix. As the periosteal-derived cells, easily harvested from intraoral procedure such as surgical extraction of impacted third molar, has the excellent potential of osteogenic capacity, tissue-engineered bone using periosteal-derived cells could be the best choice in reconstruction of maxillofacial bony defects.

Studies on the Yeasts for the Brewing of Soy Sauce(2) -Isolation, identification and classification of the yeasts in the soy sauce mash- (간장발효에 관여하는 효모에 관한 연구 (제2보) -간장 덧 중에 생육하는 효모에 대하여-)

  • Lee, Taik-Soo;Lee, Suk-Kun;Shin, Bo-Kyu
    • Applied Biological Chemistry
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    • v.13 no.2
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    • pp.171-180
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    • 1970
  • The yeasts in the soy sauce mash were isolated and identified, and they were classified by coloring with the treatment of TTC(2, 3, 5, triphenyltetrazolium chloride) agar and counted in process of time. The results obtained were as follows: a) The number of ordinary and osmophilic yeasts in 1 ml. of the soy sauce mash showed a tendency to be increased from the mashing to the mature stages and to decrease in the aging stages: $127{\times}10^3$ immediately after mashing, $83{\times}10^3$ 1 month after, $356{\times}10^3$ 3 months after, $1250{\times}10^3$ 6 months after and $65{\times}10^3$ 2 years after mashing in the case of ordinary yeasts, and 0 after mashing, $40{\times}10^3$ 1 month after, $81{\times}10^3$ 3 months after, $358{\times}10^3$ 6 month after and $23{\times}10^3$ 2 years after mashing in the case of osmophilic yeasts. b) 50 strains of yeasts were isolated from the soy sauce mash optionally in process of fermentation period, and they were identified as 7 genera and 18 species: 10 strains of Saccharomyces rouxii, 1 strain of Saccharomyces marxianus, 3 strains of Saccharomyces rosei, 1 strain of Saccharomyces fermentati, 6 strains of Saccharomyces mellis, 1 strain of Saccharomyces acidifaciens, 1 strain of Saccharomyces pastori, 3 strains of Pichia polymorpha, 2 strains of Hansenula anomala, 1 strain of Hansenula saturnus, 2 strains of Hansenula suaveolens, 5 strains of Nadsonia fulvescens, 8 strains of Debaryomyces hasenii, 1 strain of Debaryomyces nicotianae, 1 strain of Debaryomyces kloeckeri, 2 strains of Torulopsis sake, 1 strain of Torulopsis holmii and 1 strain of Candida pelliculasa. c) Distribution of yeasts according to the fermentation period was as follows: i) Saccharomyces rouxii, Saccharomyces marxianus, Saccharoymces rosei, Pichia polymorpha, Debaryomyces hansenii, Torulopsis sake, Candida pelliculosa, Debaryomyces nicotianae, Nadsonia fulvescens, Hansenula suaveolens and Hansenula saturnus were found in the early stages of fermentation. ii) Saccharomyces rouxii, Saccharomyces rosei, Saccharomyces fermentati, Saccharomyces mellis, Saccharomyces pastori, Hansenula anomala, Saccharomyces acidifaciens and Debaryomyces hansenii appeared in the mature stages. iii) Saccharomyces rouxii, Saccharomyces mellis, Nadsonia fulvescenes, Dedaryomyces hansenii, Debaryomyces kloeckeri, Torulopsis sake and Torulopsis holmii were distributed in the aging stages. d) TTC white yeasts were found in abundance in the early stages of fermentation and TTC red yeasts appeared more than 50 per cent in the mature and aging stages. e) The yeasts belonging to Saccharomyces mellis and Saccharomyces pastori were classified as TTC red yeasts, Saccharomyces acidifaciens were reel pink, Hansenula saturnus Debaryomyces kloeckeri, and Torulopsis holmii were pink, Saccharomyces marxianus and Nadsonia fulvescens were white and the others were the same as the description in the previous report. Saccharomyces rouxii ware classified for the most part as TTC red yeasts, and while some of them were red pink. f) Species of yeasts in the soy sauce mash were similar to those in the soy sauce koji, but the latter were not osmophilic and in the former case, the osmophilic yeasts were increased in process of fermentation period.

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Isolation and characterization of sigH from Corynebacterium glutamicum (Corynebacterium glutamicum의 sigH 유전자의 분리 및 기능분석)

  • Kim Tae-Hyun;Kim Hyung-Joon;Park Joon-Sung;Kim Younhee;Lee Heung-Shick
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.99-104
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    • 2005
  • Corynebacterial clones which exert regulatory effects on the expression of the glyoxylate bypass genes were isolated using a reporter plasmid carrying the enteric lacZ fused to the aceB promoter of Corynebacterium glutamicum. Some clones carried common fragments as turned out by DNA mapping technique. Subcloning analysis followed by the measurement of $\beta-galactosidase$ activity in Escherichia coli identified the region responsible for the aceB-repressing activity. Sequence analysis of the DNA fragment identified two independent ORFs of ORF1 and ORF2. Among them, ORF2 was turned out to be responsible for the aceB-repressing activity. ORF1 encoded a 23,216 Da protein composed of 206 amino acids. Sequence similarity search indicated that the ORF may encode a ECF-type $\sigma$ factor and designated sigH. To identify the function of sigH, C. glutamicum sigH mutant was constructed by gene disruption technique and the sigH mutant showed growth retardation as compared to the wild type strain. In addition, the mutant strain showed sensitivity to oxidative-stress generating agent plumbagin. This result imply that sigH is probably involved in the stress response occurring during normal cell growth.

Isolation and Structure Identification of Photosensitizer from Perilla frutescens Leaves Which Induces Apoptosis in U937 (들깻잎(Perilla frutescens)으로부터 U937 세포에 apoptosis를 유도하는 광과민성 물질의 분리 및 구조동정)

  • Ha, Jun Young;Kim, Mi Kyeong;Lee, Jun Young;Choi, Eun Bi;Hong, Chang Oh;Lee, Byong Won;Bae, Chang Hwan;Kim, Keun Ki
    • Journal of Life Science
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    • v.25 no.1
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    • pp.53-61
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    • 2015
  • In this study, we tried to separate the photosensitizer that induces apoptosis of leukemia cells (U937) from perilla leaves. Perilla leaves (Perilla frutescens Britt var. japonica Hara) are a popular vegetable in Korea, being rich in vitamins (A and E), GABA, and minerals. Dried perilla leaves were extracted with methanol to separate the photosensitizer by various chromatographic techniques. The structure of the isolated compound (PL9443) was identified by 1D-NMR, 2D-NMR, and FAB-mass spectroscopy. Absorbance of the UV-Vis spectrum was highest at 410 nm and was confirmed by the 330, 410, and 668 nm. PL9443 compound was determined to be pheophorbide, an ethyl ester having a molecular weight of 620. It was identified as a derivative compound of pheophorbide structure when magnesium comes away from a porphyrin ring. Observation of morphological changes in U937 cells following cell death induced by treated PL9443 compound revealed representative phenomena of apoptosis only in light irradiation conditions (apoptotic body, vesicle formation). Results from examining the cytotoxicity of PL9443 substance against U937 cells showed that inhibition rates of the cell growth were 99.9% with the concentration of 0.32 nM PL9443. Also, the caspase-3/7 activity was 99% against U937 cells with the concentration of 0.08 nM of PL9443 substance. The result of the electrophoresis was that a DNA ladder was formed by the PL9443. The PL9443 compound is a promising lead compound as a photosensitizer for photodynamic therapy of cancer.