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Enhanced Production of Astaxanthin by Metabolic Engineered Isoprenoid Pathway in Escherichia coli (대장균에서 이소프레노이드 생합성 경로의 대사공학적 개량에 의한 아스타잔틴의 생산성 향상)

  • Lee, Jae-Hyung;Seo, Yong-Bae;Kim, Young-Tae
    • Journal of Life Science
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    • v.18 no.12
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    • pp.1764-1770
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    • 2008
  • The goal of this study is to increase production of astaxanthin in recombinant Escherichia coli by engineered isoprenoid pathway. We have previously reported structural and functional analysis of the astaxanthin biosynthesis genes from a marine bacterium, Paracoccus haeundaensis. The carotenoid biosynthesis gene cluster involved in astaxanthin production contained six carotenogenic genes (crtW, crtZ, crtY, crtI, crtB, and crtE genes) and recombinant E. coli harboring six carotenogenic genes from P. haeundaensis produced 400 ${\mu}g$/g dry cell weight (DCW) of astaxanthin. In order to increase production of astaxanthin in recombinant E. coli, we have cloned 4-hydroxy-3-methylbut-2-enyl diphosphate reductase (lytB), farnesyl diphosphate (FPP) synthase (ispA), and isopentenyl (IPP) diphossphate isomerase (idi) in the isoprenoid pathway from E. coli and coexpressed these genes in recombinant E. coli harboring the astaxanthin biosynthesis genes. This engineered E. coli strain containing both isoprenoid pathway gene and astaxanthin biosynthesis gene cluster produced 1,200 ${\mu}g$/g DCW of astaxanthin, resulting 3-fold increased production of astaxanthin.

Assessment of Earthquake Induced Landslide Susceptibility with Variation of Groundwater Level (지하수위 변화에 따른 지진 유발 산사태의 취약섬 분석)

  • Kim, Ji-Seok;Park, Hyuek-Jin;Lee, Jung-Hyun
    • Economic and Environmental Geology
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    • v.44 no.4
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    • pp.289-302
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    • 2011
  • Since the frequency of the earthquake occurrence in Korean peninsular is continuously increasing, the possibility that massive landslides are triggered by earthquake is also growing in Korea. Previously, the landslide is known to be induced by large magnitude earthquake, whose magnitude is larger than 6.0. However, the landslide can be induced by only small magnitude earthquake, especially in the fully saturated soil. Therefore, the susceptibility of landslide caused by small magnitude earthquake in fully saturated soil is analyzed in this study. For that, the topographical and geological characteristics of the site were obtained and managed by GIS software. In the procedure of the study, slope angle, cohesion, friction angle, unit weight of soil were obtained and constructed as a spatial database layer. Combining these data sets in a dynamic model based on Newmark's displacement analysis, the landslide displacements were estimated in each grid cell. In order to check out the possibility of the earthquake induced landslides, the level of the groundwater table is varied from dry to 80% saturated soil. In addition, in order to analyze the effect of the magnitude of earthquake and distance to epicenter, four different earthquakes epicenters were considered in the study area.

Dietary protease improves growth performance, nutrient digestibility, and intestinal morphology of weaned pigs

  • Park, Sangwoo;Lee, Jeong Jae;Yang, Boung Mo;Cho, Jin Ho;Kim, Soyun;Kang, Joowon;Oh, Sejong;Park, Dong-Jun;Perez-Maldonado, Rider;Cho, Jee-Yeon;Park, Il-Hun;Kim, Hyeun Bum;Song, Minho
    • Journal of Animal Science and Technology
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    • v.62 no.1
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    • pp.21-30
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    • 2020
  • This study was conducted to evaluate effects of dietary protease (PR) on growth performance, nutrient digestibility, and intestinal morphology of weaned pigs. A total of 75 weaned pigs [7.06 ± 0.18 kg of average body weight (BW); 28 day old] were randomly allotted to 3 dietary treatments in a randomized complete block design (blocks = BW and sex): a diet based on corn and soybean meal to meet the requirement of crude protein (CP) as a positive control (PC; CP = 24.49%), a low protein diet as a negative control (NC; CP = 22.51%), and NC + 0.02% PR. The PR used in this study was a commercial product containing 75,000 protease units/g derived from Nocardiopsis prasina produced in Bacillus licheniformis. Pigs were fed the dietary treatments for 6 weeks and the diets containing 0.2% chromic oxide for the last week of this study. Blood, feces, ileal digesta, and ileum samples were collected from randomly selected two pigs in each pen on respective time points. Measurements were growth performance, apparent ileal digestibility (AID) and apparent total tract digestibility (ATTD) of dry matter (DM), CP, and energy, frequency of diarrhea, packed cell volume (PCV), and ileal morphology of weaned pigs. Pigs fed PC and PR had higher (p < 0.05) final BW, average daily gain (ADG), and gain to feed ratio (G:F) during overall experimental period than those fed NC. Pigs fed PC and PR had higher (p < 0.05) AID or ATTD of DM, CP, or energy than those fed NC. Moreover, pigs fed PR had higher ratio between villus height and crypt depth (p < 0.05) and number of goblet cells (p < 0.05) than those fed NC. Addition of PR decreased (p < 0.05) frequency of diarrhea for the first two weeks after weaning compared with PC and NC. In addition, pigs fed PR had lower (p < 0.05) PCV on d 14 after weaning than those fed PC and NC. In conclusion, addition of PR in nursery diets with a low protein level significantly improved growth performance, nutrient digestibility, and intestinal morphology of weaned pigs.

PLANT BIOCHEMISTRY OF GINSENG SAPONINS(III) Radioactive Studies (2). Sodium Acetate-U-$C^{14}$ Experiment

  • The Korean Society of Ginseng The Korean Society of Ginseng
    • Proceedings of the Ginseng society Conference
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    • 1974.09a
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    • pp.101-113
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    • 1974
  • The radioactive compound sodium $acetate-U-C^{14}$ (C-14 acetate) was administered to two- and four-year-old July and September American ginseng (Panax quinquefolium L.) plants and cuttings. The C-14 acetate uptake was approximately $99\%.$ The autoradiochromatograms suggest that the saponins(panaquilins) isolated by preparative thin-layer chromatography contained impurities, especially those isolated from the leaf and stem extracts. The root and fruit methanol extracts yielded relatively pure saponins. The large amounts of panaquilin B and its proximity to panaquilin C on preparative thin-layer plates resulted in some admixing. The average concentration $(\%$ plant dry weight) of semipurified saponins were high in the leaves $(13.8\%),$ compared to fruits $(9.8\%),\;stems\;(7.9\%)\;and\;roots\;(6.3\%).$ The average percentage of C-14 acetate incorporation into panaquilins was $4.8\%.$ The average percentage of C-14 acetate incorporation into panaquilins B and C was higher $(1.40\%\;and\;1.13\%,$ respectively) than that into panaquilin C, (d), G-1 and G-2 $(0.75\%,\;0.65\%,\;0.13\%\;and\;0.53\%,$ respectively). Panaquilin synthesis may be depending upon the part collection period and age of the plant. The average percentage of C-14 acetate incorporation into panaquilin B is high in roots $(0.58\%)\;and\;stems\;(0.48\%);$ that into panaquilins C and (d) high in leaves $(0.40\%\;and\;0.45\%,$ respectively); and that into panaquilin E high in roots and leaves $(0.55\%\and\;0.50\%,$ respectively). Panaquilin G-2 was synthesized in all parts of plants. The panaquilins appear to be biosynthesized more actively in July than September (exception-panaquilin G-l). Panaquilins B, C and G-1 may be biosynthesized more actively in four-year-old plants and panaquilins (d) and E more actively in two-year-old plants. The results from expectance with cuttings suggest that the panaquilins are synthesized de novo in the above-ground parts of ginseng plants, and that panaquilin G-l may be synthesized de novo in the leaf. It is known from the tissue culture studies that panaquilins are produced by leaf, stem and root callus tissues and callus-root cultures of American and Korean ginseng plants. Panaquilins may actively be synthesized de novo in most any cell or organ of the ginseng plants. It was verified that C-14 acetate was incorporated into the panaxadiol portions of the panaquilins of two-year-old plants (sp. act., 0.56 $m{\mu}Ci/mg$) and four-year-old plants (sp. act., 0.54 $m{\mu}Ci/mg$).

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Identification of the Marine Microorganisms Producing Bioactives -I. Isolation and Cultural Conditions of the Marine Actinomycetes No. 101 Producing Antimicrobial compounds- (생리활성물질을 생성하는 해양미생물의 동정 -I. 항미생물 물질을 생산하는 해양방선균 분리균주 No. 101의 분리 및 배양조건-)

  • CHOI Jong-Duck;PARK Uk-Yeon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.26 no.4
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    • pp.305-311
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    • 1993
  • Marine organisms were investigated to identify the marine actinomycetes that produced noble bioactive compounds. Microorganism counts range from $2.1{\times}10^3\;to\;1.2{\times}10\;CFU/g$ of marine organisms. Actinomycetes constituted 0.01 to $0.5\%$ of culturable microbial community. We identified the marine actinomycetes that produced novel bioactive compounds. During the course of screening for bioactives from the marine microorganisms, we found that the strain in sponge had antimicrobial activities. From the morphological, cultural and various physiological characteristics, this strain was identified for Actinomycetes No. 101. The optimal compositions of culture medium for Actinomycetes No. 101 were starch 30g/l as carbon source, casamino acid 10g/l as nitrogen source. The optimal pH of medium and fermentation temperature were $6.5{\sim}7.0$ and $30^{\circ}C$, respectively. Fermentation has been conducted in the marine broth at $30^{\circ}C$ for 72 hour. The yield of fermentation got about 3g as dry weight(per liter of broth). The distribution of antimicrobial activity of Actinomycetes No. 101 was screened by paper disc. The extract of cultured cell and broth inhibited the growth of Staphylococcus aureus and Bacillus subtilis, but the inhibition action was week against yeast and mold.

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Optimal Production of Xylooligosaccharide by Using Recombinant Endoxylanase from Bacillus subtilis (Bacillus subtilis 유래 재조합 endoxylanase를 이용한 xylooligosaccharide의 최적 생산)

  • Kim, Yeon-Hee;Heo, Sun-Yeon;Kim, Mi-Jin;Lee, Jae-Hyung;Kim, Young-Man;Nam, Soo-Wan
    • Journal of Life Science
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    • v.18 no.1
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    • pp.52-57
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    • 2008
  • Xylan is a major hemicellulose component of the cell walls of monocots and hardwood, representing up to 30% of the dry weight of these plants. To efficiently hydrolyze xylan, the endoxylanase gene from Bacillus sp. was expressed in B. subtilis DB431 by introducing the plasmid pJHKJ4. The total activity of the recombinant endoxylanase reached about 857 unit/ml by batch fermentation of B. subtilis DB431/pJHKJ4 in LB maltose medium. The majority (>92%) of endoxylanase was efficiently secreted into the culture medium. The recombinant endoxylanase hydrolyzed more the birchwood xylan efficiently than the other xylans. When 4 % concentration of xylan was used, the highest production of xylooligosaccharide was observed, and xylobiose and xylotriose were the major products. Optimal amount of enzyme and reaction time for producing xylooligosaccharide were found to be 10 unit and 1 hr, respectively. In addition, the temperature of $40^{\circ}C{\sim}50^{\circ}C$ gave the highest production of xylooligosaccharide. Consequently, the optimized conditions for the production of xylooligosaccharide through the hydrolysis of xylan were determined as follows: 10 unit endoxylanase, $50^{\circ}C$, 4% birchwood xylan, 1 hr reaction.

Effect of ginseng Saponin on the fatty acid composition of the lipids of Rhodotorula glutinis. (인삼 Saponin이 Rhodotorula glutinis의 지방산 조성에 미치는 영향)

  • 이시경;주현규
    • Microbiology and Biotechnology Letters
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    • v.13 no.2
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    • pp.109-114
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    • 1985
  • This study was attempted to observe the effect of crude ginseng saponin on the cell growth, lipid contents of Rhodotolula glutinis cells and fatty acid composition of the lipids from the yeast. The results obtained were as follows; The weight of dry yeast cells was increased by the addition of ginseng saponin and was noticeably increased in the treatment containing 10$^{-2}$ % ginseng saponin. With the increase of ginseng saponin concentration, lipids contents in Rhodotorula glutinis were increased but it's contents were decreased at the concentration of 10$^{-1}$ % ginseng saponin. The crude lipids of Rhodotorula glutinis were identified to be composed of palmitic, stearic, oleic, linoleic acid. Un-saturated fatty acid contents were greater than those of saturated fatty acids. Fatty acid contents were increased in the order of oleic, linoleic, palmitic and stearic acid, but palmitic acid was increased more than linoleic acid in the lipids of cells obtained at the higher than 10$^{-2}$ % ginseng saponin. As the ginseng saponin concentration was increased fatty acid contents were also increased, while they were considerably decreased in case of the addition of 10$^{-1}$ % ginseng saponin concentration.

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Phenolics Level and Antioxidant Activity of Methanol Extracts from Different Plant Parts in Youngia sonchifolia (고들빼기 부위별 메탄올 추출물의 폴리페놀 함량 및 항산화성 연구)

  • Chon, Sang-Uk;Kang, Jong-Goo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.58 no.1
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    • pp.20-27
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    • 2013
  • Proximate composition, total phenolics and total flavonoids level, DPPH radical scavenging activity, and cytotoxicity were determined in the methanol extracts of different plant parts of Youngia sonchifolia at reproductive growth stage. Crude protein and crude fat were present as the highest amount in flowers, and crude fiber in the stems and roots. The highest content of phenolics [mg ferulic acid equivalents (FAE) $kg^{-1}$ dry weight (DW)] was found in flowers (highest) and followed by leaves, stems and roots (lowest). Flavonoids [mg rutin equivalents $kg^{-1}$ DW] level, however, showed the highest in leaf extracts and lowest in root extracts. The antioxidant potential of the methanol extracts from the plants dose-dependently increased DPPH (1,1-diphenyl-2-picrylhydrazyl) free radical scavenging activity (%). DPPH radical scavenging activity were highest in root extracts ($IC_{50}=1,135.6\;mg\;kg^{-1}$) and followed by leaf, stem and flower extracts. By way of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay, methanol extracts of roots showed the highest anticancer activity on human cancer cell line Calu-6 for human pulmonary carcinoma ($IC_{50}=196.3\;mg\;kg^{-1}$) and HCT-116 for human colon carcinoma ($IC_{50}=623.6\;mg\;kg^{-1}$).

Purification and Characterization of Urushiol Induced Laccase Isoenzyme from Fomitella fraxinea (Urushiol에 의해 유도된 장수버섯 laccase isoenzyme의 정제 및 특성)

  • Choi, Han-Seok;Park, Hyo-Suk;Yeo, Soo-Hwan;Jeong, Seok-Tae;Choi, Ji-Ho;Kim, Myung-Kon
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.152-159
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    • 2010
  • The influence of urushiol, as an allergen on laccase property of Fomitella fraxinea was investigated. The enzyme production was reached to the highest level after 10 days, cultivation and the activity and mycelial biomass were increased by 2.5 and 1.5 folds, respectively, by adding urushiol in the culture medium. In liquid cultures using a Cu Mn-free medium, laccase lactivity was decreased by 3.8-9.2 folds, with similar dry cell weight. Two isoenzymes, were purified using anion exchange, hydrophobic interaction and size-exclusion chromatographies. Both isoenzymes are monomeric proteins, with $M_W$ around 67 kDa(Lac1) and 66 kDa(Lac2), and isoelectric points of 3.67 and 3.81. The optimal conditions for purified isoenzymes were found to be pH 4.5-5.0 and $30-35^{\circ}C$. Activity decreased by the addition of $Fe^{2+}$, $Mg^{2+}$, $Na^+$, and strongly inhibited by EDTA and sodium azide.

Isolation and Optimal Culture Conditions of Brevibacillus sp. KMU-391 against Black Root Pathogens Caused by Didymella bryoniae (덩굴마름병원균인 Didymella bryoniae의 생물학적 방제를 위한 길항세균의 분리 및 특성)

  • Park Sung-Min;Jung Hyuck-Jun;Kim Hyun-Soo;Yu Tae-Shick
    • Korean Journal of Microbiology
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    • v.42 no.2
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    • pp.135-141
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    • 2006
  • We isolated a bacterium which produces antifungal substances from the Sanktpeterburg soils at Russia. The iso-lated strain was identified as Brevibacillus sp. and shown a strong antifungal activity on plant pathogenic fungi. Brevibacillus sp. KMU-391 produced maximum level of antifungal substances under incubation aerobically at $30^{\circ}C$ for 48 hours in trypticase soybean broth containing 1.0% sucrose and 1.0% polypeptone at 180 rpm and initiated pH adjusted to 7.0. Precipitate of culture broth by $30{\sim}60%$ ammonium sulfate precipitation exhibited strong antifungal activity against Didymella bryoniae by dry cell weight. Butanol extract of cultured broth also shown fungal growth inhibitory activity against Botrytis cinerea KACC 40573, Botrytis fabae KACC 40962, Colletotrichum gloeosporioides KACC 40804, Colletotrichum orbiculare KACC 40808, Didymella bryoniae KACC 40669, Fusarium graminearum KACC 41040, Fusarium oxysporum KACC 40037, Fusarium oxysporum KACC 40052, Fusarium oxysporum f, sp. radicis-Iycopersici KACC 40537, Fusarium oxysporum KACC 40902, Monosporascus cannonballus KACC 40940, Phytophthora camvibora KACC 40160, Rhizoctonia solani AG-1(IA) KACC 40101, Rhizoctonia solani AG-4 KACC 40142, and Scleotinia scleotiorum KACC 41065 by agar diffusion method.