• 제목/요약/키워드: 3D co-culture

검색결과 233건 처리시간 0.025초

Optimization of Culture Medium for Lactosucrose ($^4G-{\beta}$-D-Galactosylsucrose) Production by Sterigmatomyces elviae Mutant Using Statistical Analysis

  • Lee, Jong-Ho;Lim, Jung-Soo;Song, Yoon-Seok;Kang, Seong-Woo;Prak, Chul-Hwan;Kim, Seung-Wook
    • Journal of Microbiology and Biotechnology
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    • 제17권12호
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    • pp.1996-2004
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    • 2007
  • In this study, the optimization of culture medium using a Sterigmatomyces elviae mutant was investigated using statistical analysis to increase the cell mass and lactosucrose ($^4G-{\beta}$-D-galactosylsucrose) production. In basal medium, the cell mass and lactosucrose production were 4.12 g/l and 140.91 g/l, respectively. However, because of the low cell mass and lactosucrose production, optimization of culture medium was carried out to increase the cell mass and lactosucrose production. Culture media were optimized by the S. elviae mutant using analysis of variance (ANOVA) and response surface methodology (RSM). Central composite designs using RSM were utilized in this investigation. Quadratic models were obtained for cell mass and lactosucrose production. In the case of cell mass, optimal components of the medium were as follows: sucrose 1.13%, yeast extract 0.99%, bactopeptone 2.96%, and ammonium sulfate 0.40%. The predicted maximum value of cell mass was about 5.20 g/l and its experimental value was 5.08 g/l. In the case of lactosucrose production, optimal components of the medium were as follows: sucrose 0.96%, yeast extract 1.2%, bactopeptone 3.0%, and ammonium sulfate 0.48%. Then, the predicted maximum value of lactosucrose production was about 194.12 g/l and the corresponding experimental value was about 183.78 g/l. Therefore, by culturing using predicted conditions, the real cell mass and lactosucrose production increased to 23.3% and 30.42%, respectively.

Synthesis of Copolymeric PHA by Hydrogenophaga pseudoflava and Ralstonia eutropha H16 from Vari-ous Lactones and Their Microstructural Studies (락톤류로부터 Hydrogenophaga pseudoflava와 Ralstonia eutropha H16 두 세균에 의한 공중합 PHA의 합성 및 미세구조적 특성 연구)

  • Jang, Young-Ok;Nam, Won;Choi, Mun-Hwan;Song, Jae-Jun;Yoon, Sung-Chul
    • Microbiology and Biotechnology Letters
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    • 제28권2호
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    • pp.71-79
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    • 2000
  • Two typess of copolyesters, poly(3-hydroxybutyric acid-co-4-hydroxy-butyric acid)[P(3HB-co-4HB] and poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid)[P(3HB-co-3HV)], with various monomer ratios and different degree of microstructural heterogeneity were synthesized from Ralstonia eutropha H16 and Hydrogenophaga pseudoflava by using ${\gamma}$-butyrolactone and ${\gamma}$-valerolactone, respectively. The two bacteria showed a large difference in the utilization of ${\gamma}$-butyrolactone for cell growth and PHA synthesis. H. pseudoflava synthesized P(3HB-co-4HB) copolyesters with a wide range of 4HB content from 13 to 96 mol% depending on culture conditions, whiel R. eutropha H16 was able to synthesize the copolyesters containing less than 20 mol% of 4HB. An increase in the 4HB content in the P(3HB-co-4HB) copolyesters synthesized by H. pseud-oflava induced an lowering of their melting temperatures as well as their enthalpies of fusion. The increase in the 4HB content, however, increased the rate of degradation by an extracellular P(3HB) depolymerase. NMR spectros-copy and differential scanning calorimetry showed that the P(3HB-co-4HB) copolyesters from H. pseudoflava were generally microstructurally heterogeneous. The P(3HB-co-4HB) copolyesters) synthesized by R. eutropha H16 were rather random copolymers showing less microstructural heterogeneity than those synthesized by H. pseudoflava. The NMR D value analysis suggested that the monomer distribution of the P(3HB-co-3HV) copolymers from the two bacteria were relatively random.

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Shikimate Metabolic Pathway Engineering in Corynebacterium glutamicum

  • Park, Eunhwi;Kim, Hye-Jin;Seo, Seung-Yeul;Lee, Han-Na;Choi, Si-Sun;Lee, Sang Joung;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • 제31권9호
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    • pp.1305-1310
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    • 2021
  • Shikimate is a key high-demand metabolite for synthesizing valuable antiviral drugs, such as the anti-influenza drug, oseltamivir (Tamiflu). Microbial-based strategies for shikimate production have been developed to overcome the unstable and expensive supply of shikimate derived from traditional plant extraction processes. In this study, a microbial cell factory using Corynebacterium glutamicum was designed to overproduce shikimate in a fed-batch culture system. First, the shikimate kinase gene (aroK) responsible for converting shikimate to the next step was disrupted to facilitate the accumulation of shikimate. Several genes encoding the shikimate bypass route, such as dehydroshikimate dehydratase (QsuB), pyruvate kinase (Pyk1), and quinate/shikimate dehydrogenase (QsuD), were disrupted sequentially. An artificial operon containing several shikimate pathway genes, including aroE, aroB, aroF, and aroG were overexpressed to maximize the glucose uptake and intermediate flux. The rationally designed shikimate-overproducing C. glutamicum strain grown in an optimized medium produced approximately 37.3 g/l of shikimate in 7-L fed-batch fermentation. Overall, rational cell factory design and culture process optimization for the microbial-based production of shikimate will play a key role in complementing traditional plant-derived shikimate production processes.

A Study on the Effect of Mountain Ginseng Adventitious Roots Extract (산삼부정근 추출물의 효능${\cdot}$효과에 관한 연구)

  • Yoo Yung-Geun;Joung Min-Seok;Lee Youn-Hee;Choi Jong-Wan;Kim Joong-Hoi;Paek Kee-Yoeup
    • Journal of the Society of Cosmetic Scientists of Korea
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    • 제30권3호
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    • pp.377-383
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    • 2004
  • This study reviewed the application of an extract from mountain ginseng adventitious roots which had been grown through tissue culture as a cosmetic ingredient. The mountain ginseng adventitious roots were derived from mountain ginseng callus that was induced from mountain ginseng root whose origin is estimated to date back about one hundred years ago. The adventitious roots were separated from callus and grown in a 20 L bioreactor. In order to proliferate the adventitious roots, they were cultured for 5 weeks in bioreactor. Then the harvested mountain ginseng adventitious roots were dried and extracted. For verifying skin whitening effect of an extract from the tissue-cultured mountain ginseng adventitious roots in vivo, we performed the clinical test of it. The research showed the significant skin whitening effect of a mountain ginseng adventitious roots extract and the statistical analysis showed a significant difference (p<0.0001) between sample ($2\%$ mountain ginseng adventitious roots extract) and placebo. But, some saponins showed below $10\%$ inhibitory effect of tyrosinase and melanin synthesis in B-16 melanoma. The extracts of red ginseng and ginseng which were the same concentration as the tissue-cultured mountain ginseng adventitious roots extract's showed little inhibitory effect of tyrosinase and melanin synthesis in B-16 melanoma. In DPPH test, Anti-hydroxyl radical activity of $0.5\%$ the tissue-cultured mountain ginseng adventitious roots extract was $86\%.$.

Optimization of Outdoor Cultivation of Spirulina platensis and Control ofContaminant Organisms (Spirulina platensis의 옥외배양 최적화 및 오염생물 구제)

  • Kim, Choong-Jae;Jung, Yun-Ho;Choi, Gang-Guk;Park, Yong-Ha;Ahn, Chi-Yong;Oh, Hee-Mock
    • ALGAE
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    • 제21권1호
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    • pp.133-139
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    • 2006
  • Outdoor cultivation of cyanobacterium Spirulina platensis was carried out for 40 days in a batch mode. A half concentration of the SOT based on the underground water was used as culture medium. Working volume was 5.7 tons with 0.2 m depth. During cultivation, mean water temperature, DO and light intensity were all in proper conditions for the S. platensis growth. The adjustment of pH to over 10 with Na2CO3 and addition of the 1.5% natural salt were very effective to delete contaminant organisms, Chlamydomonas moewusii and Chlorella minutissima occurred one after the other in the culture. The mean productivity of the biomass based on the dry cell weight from 14 to 25 days, after the contaminants were deleted, was 7.8 g ·m–2· d–1, which was relatively high productivity in that a half concentration of the SOT was used for the culture. Underground water used in the culture minimized contaminants invasion and addition of the 1.5% natural salt was effective to delete contaminants as well as acted as mineral supplement in outdoor cultivation of S. platensis. Harvesting using the floating activity of S. platensis was effective from mass floating in day time after overnight without agitation and illumination.

Effects of In Vitro Maturation, In Vitro Fertilization and In Vitro Culture Conditions on Bovine Embryo Production (체외성숙, 수정 및 배양에 있어서 각기 다른 배양조건들이 소 체외수정란의 생산에 미치는 효과)

  • 조성근;송상현;공일근;이효종;최상용;박충생
    • Journal of Embryo Transfer
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    • 제15권1호
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    • pp.67-75
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    • 2000
  • This study was conducted to establish the optimal conditions for in vitro embryo production using oocytes derived from follicles of slaughter-house ovaries. The ovaries of Hanwoo were obtained from a local slaughter-house. The oocytes were aspirated from visible follicles of 2~7mm in diameter. The recovered oocytes which were completely surrounded by at least 2 layers of cumulus cells and a homogeneous cytoplasmic pigmentation were used. The selected oocytes were washed 3 or 4 times with D-PBS containing 10% bovine calf serum (BCS) and matured in vitor (IVM) in Ham's F-10 supplemented with 10% BCS or 0.01 $\mu\textrm{g}$/ml epidermal growth factor(EGF) at 39$^{\circ}C$ under 5% CO2 in air for 24 hours. They were fertilizqed in vitro (IVF) with fresh sperm separated by Percoll density gradient or swim-up in TALP media. The zygotes were cultrued with or without bovine oviductal epitherial cells(BOEC) in media(HECM-6 supplemented with 11 amino acid and / or TCM-199 supplemented with 10% BCS) for 7 to 10 days. The results obtained were as follow: The cleavage rate and developmental rate to blastocyst after maturation and IVF were not significantly different between Ham's F-10 with EGF(76.0% vs. 44.0%) and BCS(75.9% vs. 43.6%)(P<0.05). The cleavage rate and development rate to blastocyst after fertilizing by swim-up or Percoll method were not signifciantly(P<0.05) different between swim-up (80.2% vs. 29.2%) and Percoll(81.9% vs. 26.5%) (P<0.05). The cleavage rate in TCM 199(80.5) was signficiantly higher than that in HECM-6 (72.0%) (P<0.05). However, developmental rate to blastocyst using TCM 199 following HECM-6 for 3 or 4 days (42.2%) was significantly higher than that in TCM-199 alone(26.7%)(P<0.05). The cleavage rate and development rate of embryos produced in vitro by exchange timing for HECM-6 media were not significantly different between in day 3(78.6% vs. 45.5%) and day 4(75.0% vs. 43.2%)(P<0.05). The cleavage rate and developmental rate to blastocysts according to co-culture system were not significantly different between with (74.2% vs. 41.4%) and without BOEC(73.95 vs. 43.5%) (P<0.05). The number of blastomere in blastocyst stage after co-culture with or without BOEC was not significantly different (106.7$\pm$5.1 and 96.6$\pm$4.0). In conclusion, the most transferable IVP embryos could be produced from Ham's F-10 medium for IVM, Percoll density gradient method for IVF sperm separation and in vitro culture in HECM-6 until day 3 or day 4, and then transferred into TCM-199 until day 9 within adequate embryo density in culture droplets after insemination.

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Effects of Fermented Achyranthes japonica Nakai, Angelica gigas Nakai, and Eucommia ulmoides Oliver Extracts on Regulation of Apoptosis in Articular Chondrocytes (Primary Chondrocytes에서 발효우슬, 당귀, 두충 복합물의 세포사멸 조절 효과)

  • Dakyung Kim;Wonhee Jo;Minhee Lee;Hyun Cheol Jeong;Sung-Jin Lee;Seunghun Lee;Jeongmin Lee
    • Journal of Applied Biological Chemistry
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    • 제66권
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    • pp.7-14
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    • 2023
  • The effects of fermented Achyranthes japonica Nakai, Angelica gigas Nakai, and Eucommia ulmoides Oliver extracts (FAAE) on regulation of inflammation and apoptosis were investigated in primary cultured rat cartilage cells. To identify the protective effects of FAAE against H2O2, cell survival was measured by MTT assay. Smad3, Collagen type I, MMP3, and MMP13 were measured by real-timpe PCR and westernbot and the inflammatory (NF-κB pathway, COX-2, iNOS) factors were determined by western blot. The apoptosis related factors (JNK, c-Fos, c-Jun, caspase 3, Bax, and Bcl-2) were determined by western blot. FAAE significantly increased the follwing: H2O2 treated cell survival, mRNA and protein expression of Smad 3, collagen type I. In addition, FAAE significantly decreased the protein expression of inflammatory and apoptosis related factors. This study suggests that FAAE have a protection effect of chondrocytes through inhibition of inflammation and apoptosis. Thus, FAAE is a therapeutic potential food componet in osteoarthritis.

Learning Methods for Effective Object Tracking in 3D Storytelling Augmented Reality (3D 스토리텔링 증강현실에서 효과적인 객체 추적을 위한 학습 방법)

  • Choi, Dae han;Han, Woo ri;Lee, Yong-Hwan;Kim, Youngseop
    • Journal of the Semiconductor & Display Technology
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    • 제15권3호
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    • pp.46-50
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    • 2016
  • Recently, Depending on expectancy effect and ripple effect of augmented reality, the convergence between augmented reality and culture & arts are being actively conducted. This paper proposes a learning method for effective object tracking in 3D storytelling augmented reality in cultural properties. The proposed system is based on marker-less tracking, and there are four modules that are recognition, tracking, detecting and learning module. Recognition module is composed of SURF and LSH, and then this module generates standard object information. Tracking module tracks an object using object tracking based on reliability. This information is stored in Learning module along with learned time information. Detecting module finds out the object based on having the best possible knowledge available among the learned objects information, when the system fails to track. Also, it proposes a method for robustly implementing a 3D storytelling augmented reality in cultural properties in the future.

Development of Shirt Design Applying Traditional Clothing Design in Northeast Asia Region - Global Fashion Cultural Products - (동북아시아 지역 전통복식 디자인을 응용한 셔츠디자인 개발 -글로벌 패션문화상품-)

  • Choi, Eunjoo
    • Fashion & Textile Research Journal
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    • 제22권4호
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    • pp.446-455
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    • 2020
  • In order to develop high value fashion culture products capable of reaching the global market, this study developed the original traditional clothing design of Northeast Asian countries according to a global perspective. This study applied the structural formality of traditional clothing to fashion cultural products. This study developed designs using Deel of Mongolia, Makap of China Korean Dang-ui / Dan-ryung, and Haori of Japan. The research methods are theoretically examined using literature on traditional clothing in Northeast Asia. First, it designed a shirt design using Adobe Illustrator CS6 and created a pattern with the Yuka program as well as applied CLO 5.0 (a 3D virtual dressing system from CLO Virtual Fashion Co.). A survey was also conducted on individual interest for the developed fashion cultural products. The design method obtained various design effects by applying the method of adding point detail parts of the clothes to the shirt that represented each country among their traditional clothes. This research can be used as basic data for the global fashion cultural products market that can contribute to maintaining the originality of each country in the global era, expanding tourism income to succeed and develop culture and tradition.

Activity of Anti-Methicillin Resistant Staphylococcus aureus Compound Derived Marine Actinomycetes and Its Synergistic Effect (해양 방선균 유래 항 Methicillin Resistant Staphylococcus aureus 물질의 활성 및 상승 효과)

  • Seong-Yun, Jeong
    • Journal of Marine Bioscience and Biotechnology
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    • 제14권2호
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    • pp.143-154
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    • 2022
  • We isolated marine actinomycetes, strain D-5 which produces anti-methicillin resistant Staphylococcus aureus (anti-MRSA) compound. Streptomyces sp. D-5 relatively grew well in the 20~25℃, pH 8.0, and NaCl 3.0%. The ethyl acetate extract of D-5 culture was separated by C18 ODS open column and reverse phase HPLC to yield anti-MRSA compound. The molecular weight of this compound was determined to be 898 by a Liquid chromatograph-mass spectrometer (LC-MS). Compared with penicillin G, this compound showed significant anti-MRSA activity. It also exhibited an inhibition zone of 26 mm at a concentration of 64 ㎍/disk and an inhibition zone of 16 mm at a concentration of 16 ㎍/disk against the MRSA KCCM 40511. Furthermore, the co-treatment of HPLC peak 5 compound and vancomycin caused a more rapid decrease in MRSA cells than each compound alone. It showed 86.8% growth inhibition activity within 12 hours at a low concentration of 50 ㎍/mL during co-treatment, and 97.1% growth in-hibition activity within 48 hours against MRSA KCCM 40511. Taken together, our results suggest that Streptomyces sp. D-5 and its anti-MRSA compound could be employed as a potent agent in MRSA infection.