• Title/Summary/Keyword: Agar concentration

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Optimal Conditions for the Production of Salt-tolerant Protease from Aspergillus sp. 101 and Its Characteristics (Aspergillus sp. 101로부터 내염성 단백분해효소 생산을 위한 최적 조건 및 특성)

  • Hwang, Joo-Yeon;Choi, Seung-Hwa;Lee, Si-Kyung;Kim, Sang-Moo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.11
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    • pp.1612-1617
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    • 2009
  • Aspergillus sp. 101 was isolated from the Korean traditional soybean paste for the production of a salt-tolerant protease. The optimal condition for the production of a salt-tolerant protease was determined with various energy sources such as carbon, nitrogen, and protein, and at different culture conditions such as temperature, pH, incubation time and NaCl concentration. The most favorable organic nitrogen sources were 2% defatted soybean flour (DSF) and soy protein isolate (SPI). Optimal pH and temperature were pH 6.0 and $25{\sim}27^{\circ}C$, respectively. Therefore, Aspergillus sp. 101 protease was a mild acid (or neutral) protease. Protease production was the highest at 0.1% concentration of $CaCO_3,\;K_2HPO_4$ and Arabicgum. Aspergillus sp. 101 could grow in culture medium at 15% NaCl concentration and produce a salt-tolerant protease even at 7% NaCl. The cell mass and protease activity of Aspergillus sp. 101 cultured in a modified medium was comparatively higher in Czapek dox and protease producing media. Hence, Aspergillus sp. 101 protease can be utilized in soy or fish sauce industry as a salt-tolerant protease starter.

Development of Osmotolerant Mutant, Candida magnoliae M26 and the Determination of the Optimum Concentrations of Carbon and Nitrogen Sources to Improve Erythritol Yield (에리스리톨의 수율 향상을 위한 Candida magnoliae의 삼투압 내성 변이균주의 개발과 탄소원 및 질소원의 최적 농도 결정)

  • 양성욱;서진호;유연우
    • KSBB Journal
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    • v.15 no.6
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    • pp.566-572
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    • 2000
  • Experimental studies were carried out to develop an osmotolerant mutant of Candida magnoliae JH and to determine the optimum concentrations of carbon and nitrogen sources to improve erythritol yield and productivity. Mutants of C. magnoliae JH were prepared by treatment with ethylmethane sulfonate, and osmotolerant mutants were isolated from the agar plate colonies containing 2.5 M KCI. Among the mutants isolated, one mutant M26 was finally selected based on erythritol yield and productivity. In shake flask culture, glucose proved to be the best carbon source and the optimum yeast extract concentration was 5 g/L based on 100 g/L glucose. The erythritol yield and productivity of mutant M26 were greater than wild type in 100 g/L glucose medium. in the fermentation experiments, erythritol production increased with increased glucose concentration, up to a limit of 250 g/L. The maximum concentration of erythritol achieved 127.5 g/L, and the yield and productivity of erythritol production were 51.0% and 0.63 g/L-h, respectively.

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EFFECTS OF SIGNAL TRANSDUCTION PATHWAY IN THE RAS-INDUCED CELLULAR TRANSFORMATION OF HUMAN EPITHELIAL CELLS IN CULTURE (인체 상피세포에서 ras-종양유전자의 발암화가 신호 전달 기작에 미치는 영향)

  • Jang, Do-Geun;Byeon, Ki-Jeong;Kim, Chin-Soo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.26 no.3
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    • pp.254-261
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    • 2000
  • The present study has attempted to look into the mechanism of ras-induced carcinogenesis in a human epithelial cell system. Human epithelial cells immortalized with Ad12-SV40 hybrid virus were used to assess carcinogenic potential of the ras-oncogene. Cells transfected with pSV2-ras showed characteristics of cellular transformation. The transformation parameters such as cell density, soft-agar colony formation, and cell aggregation were significantly increased in the cells expressing ras oncoprotein. In addition, the duration required for the appearance of foci was shortened in the ras-transfected cells. Consistent with other reports, our results demonstrated an evidence that the ras-oncogene induced the cellular transformation of human epithelial cell system. When a high concentration of glucocorticoid was added into the media, transformation process was accelerated. It is speculated that glucocorticoid may provide an advantageous environment for the proliferation of the transformed cells. The induction of the intracellular free calcium concentrations following agonist treatment was significantly lower in the transformed cells than in the control cells. These effects were more manifested in the presence of extracellular cacium, indicating that the transformation process may alter the influx pathway of extracellular calcium. The induction of $IP_3$ following agonist treatment was also lower in the transformed cells than in the control cells. Thus, it is suggested that phospholipase C-coupled pathway was down-regulated in the process of the ras-induced transformation. While the levels of $TGF-{\beta}_1$ and PAI-2 mRNAs were decreased, the level of fibronectin mRNA was increased. The results indicate that mechanism of the ras-induced transformation may be associated with the altered expressions of growth regulatory factors. The present study demonstrates an evidence that the ras-induced cellular transformation may be associated with alteration of signal transduction and growth regulatory factors. The study will contribute to improve the understanding of molecular mechanism of epithelium-derived cancers including oral cancer.

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EFFECTS OF HYDROQUINONE ON NEOPLASTIC TRANSFORMATION OF HUMAN EPITHELIAL CELLS IN CULTURE (Hydroquinone이 인체 상피세포의 발암화에 미치는 영향)

  • Sohn, Jung-Hee;Kim, Chin-Soo
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.32 no.3
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    • pp.218-228
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    • 2010
  • Components of dental resin-based restorative materials are reported to leach from the filling materials even after polymerization. Hydroquinone (HQ) is one of the major monomers used in the dental resin and is known as a carcinogen. Thus, carcinogenic risk of HQ leaching from the dental resin becomes a public health concern. The present study attempted to examine the carcinogenic potentials of HQ on the human epithelial cell, which is the target cell origin of the most of oral cancers. Cytotoxicity of HQ was observed above 50${\mu}M$ as measured by LDH assay, indicating a relatively low toxicity of this substance in human epithelial cells. The parameters of neoplastic cellular transformation such as cell saturation density, soft agar colony formation and cell aggregation were analyzed to examine the carcinogenic potential of HQ. The study showed that 2-week exposure of HQ showed the tendency of increase in the saturation density and the significant enhancement of soft agar colony formation at the highest dose, 50 ${\mu}M$ only. It is suggested that HQ has a weak potential of carcinogenicity. When cells were treated with HQ and TPA, a well-known tumor promoter, the parameters of neoplastic cellular transformation was significantly increased. This result indicates that the potential risk of carcinogenicity from HQ is largely dependent upon the presence of promoter. Exposure of 50 ${\mu}M$ HQ increased the time-dependent apoptosis as measured by the ELISA kit. This concentration coincides with a dose of neoplastic transformation, indicating a possible link between apoptosis and HQ-induced cellular transformation. Hydroquinone generated Reactive Oxygen Species (ROS) which was evidenced by the treatment of antioxidants such as trolox and N-acetyl cysteine and the GSH depleting agent, BSO. Antioxidants blocked the generation of ROS and the GSH depleting agent, BSO dramatically increased the ROS production. Since HQ is known to increase ROS production thru activation of transcriptional factor such as c-Myb and Pim-1, it is speculated that ROS generation by HQ plays a role in the activation of oncogene, which may lead to neoplastic transformation. In addition, ROS is involved in the alteration of signal transduction, which regulates the apoptosis in many cellular systems. Thus, ROS-mediated apoptosis may be involved in the HQ-induced carcinogenic processes. Protein kinase C (PKC) is known to play pivotal roles in neoplastic transformation of cells and its high expression is often found in a variety of types of tumors including oral cancer. PKC translocation of PKC-${\alpha}$ was observed following HQ exposure. Altered signaling system may also play a role in the transformation process. Taken together, HQ leached from the dental resin does not pose a significant threat as a cancer causing agent, but its carcinogenic potential can be significantly elevated in the presence of promoter. The mechanism of HQ-induced carcinogenesis involved ROS generation, apoptosis and altered signaling pathway. The present study will provide a valuable data to estimate the potential risk of HQ as a carcinogen and understand mechanism of HQ-induced carcinogenesis in human epithelial cells.

Improvement of L-Lysine Productivity by Using Cell Fusion and Immobilized System (세포융합과 고정화 시스템을 이용한 L-Lysine의 생산성 향상)

  • Ryu, Beung-Ho;Kim, Hye-Sung;Roh, Myung-Hoon;Park, Bob-Gyu;Chung, Jong-Soon;Bai, Ki-Chul
    • Korean Journal of Food Science and Technology
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    • v.21 no.1
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    • pp.154-163
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    • 1989
  • This studies were designed to improve the productivity of L-lysine by protoplast fusion and immobilized system of fusants using strains of Brevibacterium flavum ATCC 21528, Brevibacterium lactofermentum ATCC 21086 and Corynebacterium glutamicum 820. Mutants were isolated with concentration method of $300{\mu}g/ml$ penicillin-G after treatment of $250{\mu}g/ml$ N-methyl-N-nitro-N-nitrosoguanidine. B. flavum $37-2(Hos^-,\;Kan^r,\;AEC^r)$, B. lactofermentum $6-2(Ile^-,\;Val^-,\;Str^r,\;AEC^r)$ and C. glutamicum 57-5$(Met^-,\;Thr^-,\;Rif^r,\;AEC^r)$ were isolated from mutants. Protoplasts were induced by being incubated with $500{\mu}g/ml$ lysozyme of lysis solution for 6 hr and the ratio of protoplast formation and regeneration were ranging from 97-99% and 33-37%, respectively. Fusion frequencies of fusants of BBFL 21, BCFG 37 and BCLG 59 were shown in the range from $1.25{\times}10^{-6}\;to\;5.83{\times}10^{-7}$ under the optimum conditions. The fusant BBFL 21 showed the highest productivity of $411.1\;ng/ml{\cdot}hr$ L-lysine in the lysine productivity broth at $30^{\circ}C$ for 72hr. In the immobilization systems, fusant BBFL 21 was employed in various polymer matrices such as sodium alginate, polyacrylamide, agar and ${\alpha}-carrageena$. The immobilization of sodium alginate showed the highest productivity of $413\;ng/ml{\cdot}hr$ L-lysine in the batch system. Continuous fermentation of immobilization system by using tube fermentor was produced the highest productivity $416.7\;ng/ml{\cdot}hr $ L-lysine under optimum condition.

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The Factors on Somatic Embryogenesis of Soybean [Glycine max. (L.) Merrill]

  • Kim, Kyong-Ho;Kim, Hag-Sin;Oh, Young-Jin;Suh, Sug-Kee;Kim, Tae-Soo;Park, Ho-Kee;Park, Moon-Soo;Kim, Seok-Dong;Yeo, Up-Dong
    • Journal of Plant Biotechnology
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    • v.2 no.3
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    • pp.123-128
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    • 2000
  • To enhance in vitro plantlet regeneration efficiency of soybean through embryogenesis, the culture conditions such as material part and size of immature seed, 2,4-D, pH and solidifying agents for somatic embryogenesis were investigated. Somatic embryogenesis was induced from the immature embryo, immature cotyledon and embryonic axis explants of the immature seed on MS medium supplemented with 2.0 mg/L 2,4-D. The highest rate (up to 22.9%) of somatic embryogenesis was obtained from the immature cotyledon, following embryonic axis and the immature embryo. The rate varied with the developmental stages of seed. The maximum rate (25.4%) of embryogenesis was obtained from 3-4 mm length of the seed (after 25 days of flowering). The optimum concentration of 2,4-D for embryogenesis was 10 mg/L. The optimum pH was at 5.8 and solidifying agent for medium was better with 0.4% gelrite than with agar. For rapid multiplication of shoot tips from the germinating somatic embryos, they were cultured on MS medium containing 2 mg/L indole-3-butyyic acid (IBA) and 1 mg/L 6-benzyladenine (BA). After then somatic embryos with one and three cotyledons were transferred to the growth regulator free medium. The medium exhibited the higher rate (ca. 50%) of development than the multiplication medium.

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Comparison of Chemical Compositions and Antimicrobial Activities of Essential Oils from Three Conifer Trees; Pinus densiflora, Cryptomeria japonica, and Chamaecyparis obtusa

  • Lee, Jeong-Ho;Lee, Byung-Kyu;Kim, Jong-Hee;Lee, Sang-Hee;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.19 no.4
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    • pp.391-396
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    • 2009
  • The chemical compositions, and antibacterial and antifungal effects of essential oils extracted from three coniferous species, Pinus densiflora, Cryptomeria japonica, and Chamaecyparis obtusa, were investigated. Gas chromatography mass analysis of the essential oils revealed that the major components and the percentage of each essential oil were 16.66% $\beta$-phellandrene and 14.85% $\alpha$-pinene in P. densiflora; 31.45% kaur-16-ene and 11.06% sabinene in C. japonica; and 18.75% bicyclo [2,2,1] heptan-2-ol and 17.41% 2-carene in Ch. obtusa. The antimicrobial assay by agar disc diffusion method showed that $2.2{\mu}g$ of Ch. obtusa oil inhibited most effectively the growth of Escherichia coli ATCC 33312 and Klebsiella oxytoca ATCC 10031, whereas the C. japonica oil gave weak antimicrobial activity. The minimal inhibitory concentration(MIC) values for bacterial strains were in the range of 5.45-21.8 mg/ml depending on essential oils, but most Gram-negative bacteria were resistant even at 21.8 mg oil/ml. P. densiflora oil showed the most effective antifungal activity and the MIC values for Cryptococcus neoformans B42419 and Candida glabrata YFCC 062CCM 11658 were as low as 0.545 and 2.18 mg/ml, respectively. Cryp. neoformans B42419 was the most sensitive to all essential oils in the range of 0.545-2.18 mg/ml. Our data clearly showed that the essential oils from the three conifers had effective antimicrobial activity, especially against fungi.

Anti-microbial and Anticariogenic Activity of Yam and Prunella Extract against Oral Microbes (구강병인균에 대한 마와 꿀풀추출물의 항균.항우식효과)

  • Jung, Gi-Ok;Min, Kyung-Jin
    • Journal of Environmental Health Sciences
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    • v.33 no.2 s.95
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    • pp.137-144
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    • 2007
  • Yam, Prunella was stepwise extracted with hexane, chloroform, ethyl acetate, butanol, and water. Anti-microbial activity of each extract was investigated. Hexane extract was tested for anti-microbial effect on Streptocaccus mutans, one of causative factor of dental caries. Methanol extracts of 7 plants were investigated to anti-microbial effects on S. mutans KCTC 5316, P. gingivalis KCTC 5352, S. aureus KCTC 1927 by means of agar diffusion method. Methanol extract of Yam and Prunella revealed anti-microbial activity against S. mutans, P. gingivalis, and S. aureus. Also, hexane fraction of Yam revealed anti-microbial activity against S. mutans. In sequence of hexane, chloroform, ethylacetate, butanol fraction by Prunelia acted as potent anti-microbial agent on P. gingivalis. The measured MIC of hexane fraction of Yam and Prunella on S. mutans KCTC 5316 strain was 0.25 mg/ml and 0.5 mg/ml and the MIC of hexane fraction of Prunella on S. aureus was 0.5 mg/ml. The hexane fraction of Yam and Prunella suppressed viable ceil counts(VCC) of S. mutans, especially after 24 hrs. The Prunella hexane fraction suppressed VCC of S. aureus, after 12 and 24 hrs. Tested concentrations were 0.1, 0.25 and 0.5 mg/ml. the results were compared with control (0 mg/ml). The pH of S. mutans media and GTase activity were determined to evaluate the anticariogenic activity of Yam, Prunella hexane fraction. The pH were increased from 5.6 to 7.0-7.2 in concentration of 2.0 mg/ml. Yam hexane extraction revealed 35% inhibition to GTase activity and Punella inhibited 25% of GTase. These results suggest that the hexane extracts of Yam and prunella have Antibacterial activities against S. mutans, P. gingivalis, S. aureus and have preventive effect on dental caries.

Comparison of Enrichment Media of Shigella sonnei (쉬겔라 증균배지의 성능 비교)

  • In, Ye-Won;Ha, Su-Jeong;Kim, Seok-Joong;Oh, Se-Wook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.12
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    • pp.1787-1792
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    • 2011
  • The object of this study was to compare the performance of commercial enrichment media used for Shigella spp. A total of four enrichment media, Gram negative (GN) broth, Shigella broth (SB), selenite-F (SF) broth, and selenite cystine (SC) broth, were tested. When S. sonnei was inoculated into each enrichment broth at 10 cfu/mL of concentration, the highest growth was observed in Shigella broth. Morganella spp., which was not differentiated in selective agar of Shigella spp. thus can be counted as false positive, did not grow in Shigella broth in enrichment step. When S. sonnei was artificially inoculated into pork, it was mostly recovered through an enrichment process with GN broth and SF broth. However, in the case of beef, S. sonnei was mostly recovered with GN broth but largely failed with Shigella broth. Therefore, enrichment media for Shigella spp. should be selected by considering the food matrix in order to increase the chance of isolating it from foods.

Distribution Characteristics of Airborne Bacteria in Organic-Waste Resource Facilities (유기성 폐기물 자원화 시설에서 발생되는 부유 세균의 분포 특성)

  • Kim, Ki-Youn;Ko, Han-Jong;Kim, Dae-Keun
    • Journal of Environmental Health Sciences
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    • v.38 no.2
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    • pp.151-158
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    • 2012
  • Objectives: Bioaerosols released by treating organic-waste resources cause a variety of environmental and hygiene problems. The objective of this study was to investigate the distribution characteristics of the airborne bacteria emitted from a pig manure composting plant, a principal site for organic-waste resource facilities. Methods: Three types of pig manure composting plant were selected based on fermentation mode: screw type, rotary type and natural-dry type. Each site was visited and investigated on a monthly basis between September 2009 and August 2010. A total of 36 air samplings were obtained from the pig manure composting plants. The air sampling equipment was a six-stage cascade impactor. Quantification and qualification of airborne bacteria in the air samples was performed by agar culture method and identification technique, respectively. Results: The mean concentrations of airborne bacteria in pig manure composting plant were 7,032 (${\pm}1,496$) CFU $m^{-3}$ for screw type, 3,309 (${\pm}1,320$) CFU $m^{-3}$ for rotary type, and 5,580 (${\pm}1,106$) CFU $m^{-3}$ for natural dry type. The screw type pig manure composting plant showed the highest concentration of airborne bacteria, followed by the natural dry type and the rotary type. The ratio of respirable to total airborne bacteria was approximately 40-60%. The predominant genera of airborne bacteria identified were Micrococcus spp., Staphylococcus spp. and Escherichia spp. Conclusion: Monthly levels of airborne bacteria were highest in August and lowest in November regardless of fermentation mode. There was no significant correlation relationship between airborne bacteria and environmental factors such as temperature, relative humidity and particulate matters in pig manure composting plants.