• Title/Summary/Keyword: Fe-Cu-C-MnS

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Molecular cloning, purification, expression, and characterization of β-1, 4-endoglucanase gene (Cel5A) from Eubacterium cellulosolvens sp. isolated from Holstein steers' rumen

  • Park, Tansol;Seo, Seongwon;Shin, Teaksoon;Cho, Byung-Wook;Cho, Seongkeun;Kim, Byeongwoo;Lee, Seyoung;Ha, Jong K.;Seo, Jakyeom
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.4
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    • pp.607-615
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    • 2018
  • Objective: This study was conducted to isolate the cellulolytic microorganism from the rumen of Holstein steers and characterize endoglucanase gene (Cel5A) from the isolated microorganism. Methods: To isolate anaerobic microbes having endoglucanase, rumen fluid was obtained from Holstein steers fed roughage diet. The isolated anaerobic bacteria had 98% similarity with Eubacterium cellulosolvens (E. cellulosolvens) Ce2 (Accession number: AB163733). The Cel5A from isolated E. cellulolsovens sp. was cloned using the published genome sequence and expressed through the Escherichia coli BL21. Results: The maximum activity of recombinant Cel5A (rCel5A) was observed at $50^{\circ}C$ and pH 4.0. The enzyme was constant at the temperature range of $20^{\circ}C$ to $40^{\circ}C$ but also, at the pH range of 3 to 9. The metal ions including $Ca^{2+}$, $K^+$, $Ni^{2+}$,$Mg^{2+}$, and $Fe^{2+}$ increased the endoglucanase activity but the addition of $Mn^{2+}$, $Cu^{2+}$, and $Zn^{2+}$ decreased. The Km and Vmax value of rCel5A were 14.05 mg/mL and $45.66{\mu}mol/min/mg$. Turnover number, Kcat and catalytic efficiency, Kcat/Km values of rCel5A was $96.69(s^{-1})$ and 6.88 (mL/mg/s), respectively. Conclusion: Our results indicated that rCel5A of E. cellulosolvens isolated from Holstein steers had a broad pH range with high stability under various conditions, which might be one of the beneficial characteristics of this enzyme for possible industrial application.

Comparison of Physicochemical Properties and Antioxidative Activities of Sunflower Sprout According to Germination Day (발아일수에 따른 해바라기 싹의 이화학적 특성 및 항산화성 비교)

  • Roh, Kyung-Rea;Ko, Seong-Hee;Kim, Chul-Jai
    • Journal of the Korean Society of Food Culture
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    • v.27 no.1
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    • pp.66-74
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    • 2012
  • It is well known that sunflower (SF) sprout has more beneficial effects than SF seed due to increased levels of phytochemical components such as vitamins, total phenolics, and isoflavones during germination. This study investigated the physicochemical properties and antioxidative activities of SF seed during both germination and cultivation. In a proximate analysis, the water content of SF groat was 9.17% and then increased to 15.32% on the 11th day after seeding. On a dry weight basis, crude fat content decreased while the contents of carbohydrates, crude protein, and crude ash increased. As cultivation proceeded, the contents of minerals were in decreasing order of K, Mg, Ca, Na, Zn, Fe, Mn, and Cu, in which SF sprout cultivated for 9 or 11 days contained the highest mineral contents. Though vitamin C was not detected on SF groat, the content of vitamin C continuously increased up to the 5th day of cultivation and then decreased gradually. Vitamin E content in SF groat was higher than that in SF sprout. It was also found that the vitamin E content in SF sprout was the highest on the 5th day of cultivation. Daidzin was not detected in SF groat, but its concentration reached a maximum on the 5th day of cultivation in SF sprouts. Furthermore, higher amounts of daidzein were observed on the 3rd, 5th, and 9th days of cultivation. The highest total isoflavone content was observed on either the 3rd or 5th day of cultivation. The highest content of total phenolics was observed on the 5th of cultivation. When DPPH radical and peroxyl radical scavenging activities of SF sprout were measured in order to measure antioxidant efficacy, it was found that 5 day-cultivated SF sprout had the highest scavenging activities. In conclusion, SF sprout cultivated for 9 or 11 days was found to be a good source of minerals. Furthermore, the fifth-day after seeding was the optimal time for the production of SF sprout with effective natural antioxidant activity and high amounts of functional components such as vitamins, total phenolics, and isoflavones.

Effect of Pre-plant pH Change and Leaching on Crop Growth, Nutrient Uptake and Yield of Strawberry(Fragaria ${\times}$ ananassa Duchesne) 'Nyoho' (정식전 pH 변화를 수반한 토양 용탈이 딸기의 생육, 무기염 흡수 및 수확량에 미치는 영향)

  • Choi Jong Myung;Jung Suck Kee;Lee Young Bok
    • Journal of Bio-Environment Control
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    • v.13 no.4
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    • pp.233-239
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    • 2004
  • This research was conducted to determine the effect of soil pH change followed by leaching with ground water on crop growth, yield and nutrient uptake of strawberry. Treatments were leaching with ground water (A), hydrated lime application of dissolved portion in $500g{\cdot}L^{-1}$ solution followed by leaching (B), application of $Al_2SO_4$ solution with the concentration of $0.75g{\cdot}L^{-1}$ followed by leaching (C), pH elevation by (B) followed by pH lowering by (C), then leaching (D). Soil samples taken before experiment had 6.0 in soil pH. But the pH of all treatments taken after pH changes and leaching were in the range from 5.8 to 6.2 without statistical differences among treatments. Electrical conductivity of soils taken before experiment was $1.82dS{\cdot}m^{-1}$ and those of (A), (B), (C), and (D) were 1.65, 1.72, 1.71 and $1.83dS{\cdot}m^{-1}$, respectively. The plant height in treatment (C) was 20.4 cm at 80 days after transplanting which was the highest among treatments tested. The other growth characteristics did not show statistical differences among treatments. Tissue contents of Fe, Mn, Zn, Cu and B in treatment (C) were higher than those of other treatments at 80 days after transplanting. The treatment (C) had the highest number of special sizes and fruit weight per 50 plants such as 137 and 3.784 g, respectively. The treatment also had 3.123 kg in total fruit weight and 2,489 kg in marketable fruit weight per 10 are. The treatment (B) also had good results such as 2,820 kg in marketable fruit weight per 10 are.

Purification and Enzymatic Characteristics of the Bacillus pasteurii Urease Expressed in Escherichia coli (Escherichia coli에서 발현된 Recombinant Bacillus pasteurii Urease의 정제 및 효소학적 특성)

  • 이은탁;김상달
    • Microbiology and Biotechnology Letters
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    • v.20 no.5
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    • pp.519-526
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    • 1992
  • The gene coding for urease of alkalophilic Bacillus pasteurii had been cloned in Escherichia coli previously. The urease protein was purified 63.1-fold by TEAE-cellulose, DEAE-Sephadex A-50, Sephadex G-150 and Sephadex G-200 chromatographies with a 7.3% yield from the sonicated fluid of the E. coli HB1Ol(pBUll) encoding B. pasteurii urease gene. The ureases of E. coli (pBUll) and B. pasteurii possessed as a $K_m$ for urea, 42.1 mM and 40.4 mM, respectively. They hydrolyzed urea with $V_{max}$ of 86.9$\mu$mol/min and 160$\mu$mol/min, respectively. Both ureases were composed with four subunits (Mrs 67,000) and a subunit (Mr 20,000). The molecular weight of both native enzymes was Mr 280,OOO$pm$10,000 determined by gel filtration chromatography and Coomassie blue staining of the subunits. The optimal reaction pH of both ureases were pH 7.5. The ureases were stabled in pH 5.5-10.5. The optimal reaction temperature of both ureases were $60^{\circ}C$, and the ureases were stable for an hour at $50^{\circ}C$, 40min at $60^{\circ}C$ and 10 min at $70^{\circ}C$ The activity of both enzymes were inhibited completely by $Ag^{2+}$, $Hg^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and were inhibited 60% by CoH, 30% by $Fe^{2+}$ and 10% by $Pb^{2+}$. However it was increased by the addition of $Sn^{2+}$, $Mn^{2+}$, $Mg^{2+}$ at concentration of $1{\times}10^{-3}$M. Both ureases were inhibited completely by p-CMB and acetohydroxamic acid. The urease expressed in E. coli (pBU11) was inhibited 70% by SDS. The urease of B. pasteurii was inhibited 40% by hydroxyurea, whereas the recombinant urease of E. coli strain was inhibited 17%. Both enzymes were not inhibited by cyclohexanediaminetetraacetic acid (CDTA) and ethylendiaminetetraacetic acid (EDTA).

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Dyeing Properties of Yerba Mate Tea on the Fabrics (마테차를 이용한 직물의 염색성)

  • Yoo, Hye-Ja;Jeon, Sung-Taek
    • Journal of the Korean Society of Clothing and Textiles
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    • v.36 no.4
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    • pp.412-421
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    • 2012
  • The yerba mate that grows in the subtropical forests of South America is an evergreen tree within the Aquifoliaceae family. The yerba mate (Ilex paraguariensis) tea is habitually used as a drink in Argentina, Paraguay, Uruguay, and Southern Brazil. A wild mate tree needs about 25 years to fully grow to 15 a height of meters; however, they only grow to a height of 3-5 meters when cultivated. The leaves are 7-11 cm long and 3-5cm wide with a serrated margin. Yerba mate tea-based beverages are made from the leaves and stems of the mate tree. It is known that they contain a rich content of antioxidants and polyphenol, vitamins, amino acids, minerals and colorants. Dyeing properties of a mate plant in the silk, wool, linen and cotton fabrics were investigated. Constituents of colorant of mate tea are chlorophyll and carotinoid; in addition, it has more minerals such as (Fe, Ca, Mn, Mg, Na, K, Zn, and Cu) than green tea. Mate tea colorants showed good affinity to silk and wool fabrics. It was found that the optimum condition for dyeing was 30 minutes for dyeing time, 7 for pH of dyebath, and at a dyeing temperature of $80^{\circ}C$. The maximum absorbance wavelength (${\lambda}_{max}$) of silk and wool fabrics dyed with mate tea were at 420-440nm; however, that of linen and cotton fabrics were at 400nm. K/S values of fabrics dyed under optimum conditions were 1.979 for silk fabrics, 1.541 for wool fabrics, 0.551 for linens and 0.465 for cotton fabrics. Munsell hue values of dyed fabrics measured 3.1Y-6.4GY for silk, 1.4Y-8.3Y for wool, 5.5Y-3.7GY for linen, and 5.3Y-1.3GY for cotton. All dyed fabrics showed hues in-between greenish yellow and greenyellow. Colorfastness to rubbing, perspiration and dry-cleaning of dyed fabrics with mate tea were very good; however, the grades of colorfastness to washing of dyed linen and cotton fabrics were poor, and colorfastness to light were poor.

Exposure Assessments of Environmental Contaminants in Ansim Briquette Fuel Complex, Daegu(II) - Concentration distribution and exposure characteristics of TSP, PM10, PM2.5, and heavy metals - (대구 안심연료단지 환경오염물질 노출 평가(II) - TSP, PM10, PM2.5 및 중금속 농도분포 및 노출특성 -)

  • Jung, Jong-Hyeon;Phee, Young-Gyu;Lee, Jun-Jung;Oh, In-Bo;Shon, Byung-Hyun;Lee, Hyung-Don;Yoon, Mi-Ra;Kim, Geun-Bae;Yu, Seung-do;Min, Young-Sun;Lee, Kwan;Lim, Hyun-Sul
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.25 no.3
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    • pp.380-391
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    • 2015
  • Objectives: The objective of this study is to assess airborne particulate matter pollution and its effect on health of residents living near Ansim Briquette Fuel Complex and its vicinities. Also, this study measured and analyzed the concentration of TSP, $PM_{10}$, $PM_{2.5}$, and heavy metals which influences on the environmental and respiratory disease in Ansim Briquette Fuel Complex, Daegu, Korea. Methods: In this study, we analyzed various environmental pollutants such as particulate matter and heavy metals from Ansim Briquette Fuel Complex that adversely affected local residents's health. In particular, we verified the concentration distribution and characteristics of exposure for TSP, $PM_{10}$, and $PM_{2.5}$ among particulate matters, and heavy metals(Cd, Cr, Cu, Mn, Ni, Pb, Fe, Zn, and Mg). In that regard, the official test method on air pollution in Korea for analysis of particulate matter and heavy metal in atmosphere were conducted. The large capacity air sampling method by the official test method on air pollution in Korea were applied for sampling of heavy metals in atmosphere. In addition, we evaluated the concentration of seasonal environmental pollutants for each point of residence in Ansim Briquette Fuel Complex and surrounding area. The sampling measured periods for air pollutants were from August 11, 2013 to February 21, 2014. Furthermore, we measured and analyzed the seasonal concentrations(summer, autumn and winter). Results: The average concentration for TSP, $PM_{10}$, and $PM_{2.5}$ by direct influence area at Ansim Briquette Fuel Complex were 1.7, 1.4 and 1.9 times higher than reference region. In analysis results of seasonal concentrations for particulate matter in four direct influence and reference area, concentration levels for winter were generally somewhat higher than concentrations for summer and autumn. The average concentrations for Cd, Cr, Mn, Ni, Pb, Fe, and Zn in direct influence area at Ansim Briquette Fuel Complex were $0.0008{\pm}0.0004{\mu}g/Sm^3$, $0.0141{\pm}0.0163{\mu}g/Sm^3$, $0.0248{\pm}0.0059{\mu}g/Sm^3$, $0.0026{\pm}0.0011{\mu}g/Sm^3$, $0.0272{\pm}0.0084{\mu}g/Sm^3$, $0.4855{\pm}0.1862{\mu}g/Sm^3$, and $0.3068{\pm}0.0631{\mu}g/Sm^3$, respectively. In particularly, the average concentrations for Cd, Cr, Mn, Ni, Pb, Fe, and Zn in direct influence area at Ansim Briquette Fuel Complex were 1.9, 3.6, 2.1, 1.9, 1.4, 2.6, and 1.2 times higher than reference area, respectively. The continuous monitoring and management were required for some heavy metals such as Cr and Ni. Moreover, the average concentration in winter for particulate matter in direct influence area at Ansim Briquette Fuel Complex were generally higher than concentrations in summer and autumn. Also, average concentrations for TSP, $PM_{10}$, and $PM_{2.5}$ were from 1.5 to 2.0 times, 1.2 to 1.8 times, and 1.1 to 2.3 times higher than reference area, respectively. In results for seasonal atmospheric environment, TSP, $PM_{10}$, $PM_{2.5}$, and heavy metal concentrations in direct influence area were higher than reference area. Especially, the concentrations in C station were a high level in comparison with other area. Conclusions: In the results, some particulate matters and heavy metals were relatively high concentration, in order to understand the environmental pollution level and health effect in surrounding area at Ansim Briquette Fuel Complex. The concentration of some heavy metals emitted from direct influence area at Ansim Briquette Fuel Complex were relatively higher than reference area. In particular, average concentration for heavy metals in this study were higher than average concentrations in air quality monitoring station for heavy metal for 7 years in Deagu metropolitan region. Especially, the residents near Ansim Briquette Fuel Complex may be exposed to the pollutants(TSP, $PM_{10}$, $PM_{2.5}$, and heavy metals, etc) emitted from the factories in Ansim Briquette Fuel Complex.

Studies on the Nutritional Value of Elderberry (Sam bucus canadensis) Fruits (Elderberry(Sam bucus canadensis) 과실(果實)의 영양가(營養價)에 대(對)하여)

  • Park, Kyo Soo
    • Journal of Korean Society of Forest Science
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    • v.67 no.1
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    • pp.42-49
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    • 1984
  • The elderberry was known to the ancients for its medicinal properties, and in Europe the inner back was formerly administered as a cathartic. The flowers contain a voletile oil, and serve for the distilation of elder-lower water, used in confectionery, perfumes and lotions. The leaves are employed to impart a green colour to fat and oil, and the leaves and bark emit a sickly odour, believed to be repugnant to insect. With its unique flavor and natural food colour, commercial processing companies used the fruit mainly in the making for jam, jelly, pies, juice, and wines. Its vitamin-C content is reported by Andross (1941) as 25-30mg/100g. Harvesting and processing have been mechanized to some extent. However, the cotains with nutritional value has not been reported yet. In the present study the various contains with nutritional value in the fresh elderberry juice is reported by the quantitative analysis. In this study results obtained can be summarized as follows. 1) The fresh elderberry juice contained following mineral elements; calcium 0.012%, magnesium 0.023%, potassium 0.10%, sodium 0.0019%, iron 0.0009%, cobalt 0.0002%, zinc 0.0004%, copper 0.0001%, phosphorus 0.036%, manganese 0.0006%, iodide $1{\mu}g/g$. 2) Five kinds of vitamines were also found ; vitamin-$B_1$ $0.1{\mu}g/g$, vitamin-$B_2$ $0.5{\mu}g/g$, vitamin-C 0.3mg/g, niacin $14{\mu}g/g$, choline chloride 0.3mg/g. 3) Fresh elderberry juice also contains crude protein 1.10%, fat 0.26%, carbohydrate 6.9%, pectin 0.76%, tannin 0.89%, ash 0.80%, water 90.9% and 34.3 cal/100g. 4) The absorption spectrum of the purplishblack color of fresh elderberry juice has a peak between 523-530mm.

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Evaluation of Slurry, Urine and Fermented Liquid Manure at Pig Farms in the Jeju Area Regarding Chemical Composition and Pollution Level (제주지역 양돈장에서 생산된 액비의 비료성분 및 오염도 평가)

  • Kim, M.C;Song, S.T.;Hwang, K.J.
    • Journal of Animal Science and Technology
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    • v.46 no.3
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    • pp.469-478
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    • 2004
  • Pig slurry, urine and fermented liquid manure were evaluated measuring electrical conductivity (EC), dry matter (DM) contents and other components. Samples were collected during a period from February to April, 2001: slurry samples from 70 storage tanks, urine samples from 19 and 20 fermented liquid manure samples from 109 pig farms in Jeju. Samples were analyzed for pH, EC, DM, $NH_4$-N, K, P, Ca, Mg, Na, BOD, S-S, Cd, Cr, Cu, Fe, Mn, Ni, Pb, Zn and OM. Relationships between EC or DM and various mineral and $NH_4$-N contents were examined using linear regression. EC, DM, all mineral contents and $NH_4$-N were higher in slurry samples than in urine or fermented liquid manure. EC, $NH_4$-N, Ca, Mg and Na concentration was similar in urine and fermented liquid manure, but K and DM values in liquid manure were twice as much as in urine. P concentration was higher in urine than in liquid manure. $NH_4$-N and Na in slurry, $NH_4$-N in urine and $NH_4$-N, P and Ca in fermented liquid manure were highly correlated with EC, while $NH_4$-N, P, Ca and Mg in slurry, $NH_4$-N and Ca in urine, and K, P, Ca, Mg, and Na in fermented liquid manure with DM (P<0.05). BOO in slurry (22,520 mg/mL) was higher than that in urine (4,763) and fermented liquid manure (2,701). Results indicated that slurry is a better fertilizer source than urine or fermented liquid manure. However, slurry may pollute soil more than urine or fermented liquid manure when applied to land, although the levels are not above the permit.

Studies on the Constituents of Gynostemma pentaphyllum Makino (덩굴차(Gynostemma pentaphylum Makino)의 성분에 관한 연구)

  • Lee, Heon-Ok;Ko, Young-Su
    • Korean journal of food and cookery science
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    • v.6 no.4 s.13
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    • pp.69-83
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    • 1990
  • Gynostemma pentaphyllum Makino is a kind of perennial liana plant belonging to the Cucurbitaceae family. Since it was first discovered and reported to the Japanese academy in 1977, it came to be widely known to China and Southeast Asia areas as well as Japan. In Korea its name began to appear in the first half of 1980's, and it is now being nationwidely cultivated and popularized as tea stuffs. Gynostemma pentaphyllum Makino has remarkably strong propagation power. In addition. since it contains a great quantity of beneficial components, especially saponin it is believed to have excellent medical effects, too. This study was undertaken to examine the value of Gynostemma pentaphyllum Makino as a health-promotint tea, by investigating the components creating the flaver of tea and the saponin ingredient making medical effects as well as by inspecting whether Gynostemma pentaphyllum Makino products show different component arrangements according to their growing districts. As raw materials of the experiment, three kinds of Gynostemma pentaphyllum Makino samples produced in Suwon, Geochang and Uleungdo were taken. To accomplish the formerly stated goals, the contained quantity of the proximate composition, free sugar, reducing sugar, free amino acid, minerals, tannin, caffeine and vitamin C were measured respectively and compared between the samples, while the saponin content was compared with heat extracted from panax ginseng. The results of the experiment are as follows: 1. In the case of the proximate composition, the crude fat content was the highest in the Gymostemma pentaphyllum Makino material from Geochang (1.62%), the second in that from Suwon (1.56%), and the lowest in that from Uleungdo (1.0%). In addition the Geochang: produced sample had the greatest quantity of the crude protein and ash contents: the order of the crude Protein content was the Geochang produced (17.83%), the Suwon-produced (15.87%), and the Uleungdo-produced(12.28%), while that of the ash content was the Geochang-produced (14.80%), the Uleungdo-produced(10.17%), and the Suwon-produced(9.34%). 2. As for the reducing sugar contents, the Suwon-produced scored the highest of the three (3.3%), while the Geochang-produced was 1.3% and the Uleungdo-produced 0.5%. The total content of free sugar was 1.07% (the Suwon-produced), and 0.49% (the Geochang-produced) respectively but the sample from Uleungdo showed almost no free sugar content. The contained quantity of fructose and glucose was the highest among the kinds of free sugar both in the Suwon-produced and in the Geochang-produced. 3. The content of amino acid was the highest in the Suwon-produced(1.41%), the second in the Geochang-produced(1.37%), and the lowest in the Uleungdo-produced(0.53%). In the experiment, sixteen kinds of amino acid were extracted-Asp. Thr. Ser. Glu. Gly. Ala. Val. Het. Ileu. Leu. Tyr. Phe. Lys. His. Arg. Try. All of them except glutamic acid and methionine showed the highest quantity score in the Suwon-produced, while the glutamic acid content was the higest in the Uleungdo-produced and the methionine content in the Geochang-produced. The sequential arrangement of the sixteen contents according to their magnitude ranged from glycine, aspartic acid, and glutamic acid (the highest) to tryptophan, serine and lysine (the lowest). 4. Ten kinds of mineral were detected-Ca, Mn, Cd, K, Na, Pb, Mg, Fe, Zn, Cu. Among them, the content of Cd, Na, Mg, Zn and Fe was the highest in the Geochang-produced and that of K in the Suwon-produced. 5. The Geochang-produced materials contained much a larger quantity of tannin (6.3%) than the Suwon-produced (2.6%). Neither caffeine nor vitamin C was detected in the three kinds of materials. 6. In the case of the saponin content the Geochang-produced showd 2.39%, the Uleungdo-produced 1.77% and the Suwon-produced 1.49% respectively. However, it was found also that the saponin content of Gymostemma pentaphyllum Makino was the same kind as that of panax ginseng.

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Physico-Chemical Properties of Aggregate By-Products as Artificial Soil Materials (골재 부산물의 용토재 활용을 위한 특성 분석)

  • Yang, Su-Chan;Jung, Yeong-Sang;Kim, Dong-Wook;Shim, Gyu-Seop
    • Korean Journal of Soil Science and Fertilizer
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    • v.40 no.5
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    • pp.418-428
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    • 2007
  • Physical and chemical properties of the aggregate by-products including sludge and crushed dust samples collected from the 21 private companies throughout the country were analyzed to evaluate possible usage of the by-products as artificial soil materials for plantation. The pH of the materials ranged from 8.0 to 11.0. The organic matter content was $2.85g\;kg^{-1}$, and the total nitrogen content and available phosphate content were low as 0.7 percents and $12.98mg\;kg^{-1}$, respectively. Exchangeable $Ca^{2+}$, $Mg^{2+}$, $K^+$, and $Na^+$ were 2.29, 0.47, 0.02 and $0.05cmol\;kg^{-1}$, respectively. Heavy metal contents were lower than the limits regulated by environmental law of Korea. Textural analysis showed that most of the materials were silt loam with low water holding capacity ranged from 0.67 to 7.41 percents, and with low hydraulic conductivity ranged from 0.4 to $2.8m\;s^{-1}$. Mineralogical analysis showed that the aggregate by product materials were mostly composed of silicate, alumina and ferric oxides except calcium oxide dominant materials derived from limestones. The primary minerals were quartz, feldspars and dolomites derived from granite and granitic gneiss materials. Some samples derived from limestone material showed calcite and graphite together with the above minerals. According to the result, it can be concluded that the materials could be used as the artificial soil material for plantation after proper improvement of the physico-chemical properties and fertility.