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Characterization of Extracellular Cholesterol Oxidase Produced from Soil Microorganism (토양 미생물로부터 생산된 Extracellular Cholesterol Oxidase의 특성)

  • Park, Jeong-Su;Jeong, Jong-Moon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.11
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    • pp.1507-1514
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    • 2008
  • Cholesterol oxidase catalyses the conversion of cholesterol to 4-cholesten-3-one. This enzyme has been used for clinical assay of human serum cholesterol and for reduction of cholesterol level in foods and feeds. In order to search the microorganism which has a high extracellular and stable activity of cholesterol oxidase, soil microorganisms were screened. As a result, the one with the highest extracellular cholesterol oxidase activity was obtained and named as the BEN 115. The BEN 115 strain was identified as one of the Nocardia species based on our taxonomic studies. The cholesterol oxidase from this strain was shown to have two bands of extracellular proteins on SDS-PAGE and Western blot. Their molecular masses were estimated to be about 55 and 57 kDa, respectively. In addition, this cholesterol oxidase was considerably stable at the broad range of pH $3.5{\sim}9.5$ and at the temperature of $25{\sim}55^{\circ}C$. The optimum pH and temperature of this cholesterol oxidase were pH 5.5 and $35^{\circ}C$, respectively. The activity of extracellular cholesterol oxidase could be enhanced 1.6 to 2.0 folds by the addition of nonionic detergent such as Triton X-114, Triton X-100, or Tween-80 into the culturing broth. The substrate specificities against campesterol, sitosterol and stigmasterol were measured to be 50%, 50%, and 27%, respectively, compared to the cholesterol. These results suggest that Nocardia sp. BEN 115 may be useful as a microbial source of cholesterol oxidase production.

Immobilization of Cyclodextrin Glucanotransferase for Production of 2-O-\alpha-D-Glucopyranosyl L-Ascorbic Acid. (2-O-\alpha-D-Glucopyranosyl L-Ascorbic acid 생산을 위한 Cyclodextrin glucanotransferase의 고정화)

  • 성경혜;김성구;장경립;전홍기
    • Microbiology and Biotechnology Letters
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    • v.31 no.4
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    • pp.368-376
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    • 2003
  • Cyclodextrin glucanotransferase (CGTase) from Paenibacillus sp. JB-13 was immobilized on various carriers by several immobilization methods such as ionic binding, covalent linkage and ultrafiltration to improve the process performance. The ultrafiltration and covalent linkage with CNBr-activated sepharose 4B were found as the best method for immobilization of CGTase. The ability of CGTase immobilization onto CNBr-activated sepharose 4B was as high as 18,000 units/g resin when the conditions was as follows: contact time 9 hrs at $37^{\circ}C$, pH 6.0, 100 nm and enzyme loading 24,000 units/g resin. The optimum conditions for production of 2-O-$\alpha$-D-Glucopyranosyl L-Ascorbic acid by immobilized CGTase turned out to be: pH 5.0, temperature $37^{\circ}C$, 20% substrate solution containing 8% (w/v) of soluble starch and 12% (w/v) of L-ascorbic acid sodium salt, 100 rpm, far 25 hrs and with 800 units of immobilized CGTase/ml substrate solution. Moreover the CGTase activity could be stably maintained for 8 times of repetitive reactions after removing products by ultrafiltration through YM 10 membrane.

Measurement of the Intestinal Digestibility of Rumen Undegraded Protein Using Different Methods and Correlation Analysis

  • Wang, Y.;Zhang, Y.G.;Liu, Xiaolan;Kopparapu, N.K.;Xin, Hangshu;Liu, J.;Guo, Jianhua
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.10
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    • pp.1454-1464
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    • 2015
  • Four methods were adopted, including the mobile nylon bag (MNB) method, modified three-step in vitro (MTS) method, original three-step in vitro (OTS) method, and acid detergent insoluble nitrogen (ADIN) estimating method, to evaluate the intestinal digestibility of rumen undegradable protein (DRUP) of 10 types of concentrates and 7 types of roughages. After correlation analysis to determine the DRUP values using the MNB, MTS, OTS, and ADIN methods, the study aimed to find out appropriate methods to replace the MNB method due to its disadvantages such as high price, long time period, and use of a duodenal T-fistula. Three dairy cows with a permanent ruminal fistula and duodenal T-fistula were used in a single-factor experimental design. The results showed that the determined DRUP values using the MNB method for soybean meal, cottonseed meal, rapeseed meal, sunflower meal, corn germ meal, corn, rice bran, barley, wheat bran, corn fiber feed, Alfalfa (Zhao dong), Alfalfa (Long mu 801), Alfalfa (Long mu 803), grass (North), Grass (Inner Mongolia), corn silage and corn straw were 98.13%, 87.37%, 88.47%, 82.60%, 75.40%, 93.23%, 69.27%, 91.27%, 72.37%, 79.03%, 66.72%, 68.64%, 73.57%, 50.47%, 51.52%, 54.05%, and 43.84%, respectively. The coefficient of determination ($R^2=0.964$) of the results between the MTS method and the MNB method was higher than that ($R^2=0.942$) between the OTS method and the MNB method. The coefficient of determination of the DRUP values of the concentrates among the in vitro method (including the MTS and OTS methods) and the MNB method was higher than that of the roughage. There was a weak correlation between the determined DRUP values in concentrates obtained from the ADIN method and those from the MNB method, and there was a significant correlation (p<0.01) between the determined DRUP values of the roughage obtained from the MNB method and those obtained from ADIN method. The DRUP values were significantly correlated with the nutritional ingredients of the feeds. The regression equation was DRUP =100.5566+0.4169CP - 0.4344SP - 0.7102NDF - 0.7950EE ($R^2=0.8668$, p<0.01; CP, crude protein; SP, soluble protein; NDF, neutral detergent fiber; EE, ether extract). It was concluded that both the MTS method and the OTS may suitable to replace the MNB method for determining the DRUP values and the former method was more effective. Only the ADIN method could be used to predict the values of the roughages but conventional nutritional ingredients were available for all of the samples' DRUP.

Insecticide Susceptibility of Western Flower Thrip, Frankliniella occidentalis (Thysanoptera: Thripidae) on Horticultural Crops in Gyeonggi Area (경기지역 원예작물 꽃노랑총채벌레 약제 감수성)

  • Lee, Young-Su;Lee, Hee-A;Lee, Hyun-Ju;Hong, Soon-Sung;Kang, Chang-Sung;Choi, Yong-Seok;Kim, Hyeong-Hwan;Jang, Myoung-Jun
    • Korean journal of applied entomology
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    • v.56 no.2
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    • pp.179-186
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    • 2017
  • This study was conducted to monitor the insecticidal susceptibility of western flower thrip, Frankliniella occidentalis, which damage horticultural crops in the Gyeonggi area. Bioassays were conducted under laboratory and greenhouse conditions by using the recommended concentrations of commercial insecticides being used for the control of thrips. Neonicotinoid insecticides, especially acetamiprid (8%) soluble powder (SP), clothianidin (8%) SC, imidacloprid (10%) wettable powder (WP) and thiacloprid (10%) water dispersible granule (WG) were not toxic to F. occidentalis collected from horticultural crops. However, F. occidentalis collected from vegetable greenhouses was extremely susceptible to acetamiprid + spinetoram (6 + 4%) suspension concentration (SC), clothianidin + spinetoram (6 + 4%) SC and methoxyfenozide + spinetoram (6 + 4%) SC, which resulted in over 90% control of thrips. In the greenhouse test, spinetoram (5%) WG, which caused 100% F. occidentalis mortality in the laboratory test, showed 87.4, 88.0, and 98.3% control at 3, 6, and 9 days after treatment, respectively, while imidacloprid (10%) WP showed below 44% control. From the results of this study, spinosin insecticides, such as more than over 4 and 10% of spinetoram and spinosad, and pyrrole insecticide, such as 5% chlorfenapyr, are recommended for the effective control of F. occidentalis.

Tribological study on the thermal stability of thick ta-C coating at elevated temperatures

  • Lee, Woo Young;Ryu, Ho Jun;Jang, Young Jun;Kim, Gi Taek;Deng, Xingrui;Umehara, Noritsugu;Kim, Jong Kuk
    • Proceedings of the Korean Vacuum Society Conference
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    • 2016.02a
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    • pp.144.2-144.2
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    • 2016
  • Diamond-like carbon (DLC) coatings have been widely applied to the mechanical components, cutting tools due to properties of high hardness and wear resistance. Among them, hydrogenated amorphous carbon (a-C:H) coatings are well-known for their low friction properties, stable production of thin and thick film, they were reported to be easily worn away under high temperature. Non-hydrogenated tetrahedral amorphous carbon (ta-C) is an ideal for industrial applicability due to good thermal stability from high $sp^3$-bonding fraction ranging from 70 to 80 %. However, the large compressive stress of ta-C coating limits to apply thick ta-C coating. In this study, the thick ta-C coating was deposited onto Inconel alloy disk by the FCVA technique. The thickness of the ta-C coating was about $3.5{\mu}m$. The tribological behaviors of ta-C coated disks sliding against $Si_3N_4$ balls were examined under elevated temperature divided into 23, 100, 200 and $300^{\circ}C$. The range of temperature was setting up until peel off observed. The experimental results showed that the friction coefficient was decreased from 0.14 to 0.05 with increasing temperature up to $200^{\circ}C$. At $300^{\circ}C$, the friction coefficient was dramatically increased over 5,000 cycles and then delaminated. These phenomenon was summarized two kinds of reasons: (1) Thermal degradation and (2) graphitization of ta-C coating. At first, the reason of thermal degradation was demonstrated by wear rate calculation. The wear rate of ta-C coatings showed an increasing trend with elevated temperature. For investigation of relationship between hardness and graphitization, thick ta-C coatings(2, 3 and $5{\mu}m$) were additionally deposited. As the thickness of ta-C coating was increased, hardness decreased from 58 to 49 GPa, which means that graphitization was accelerated. Therefore, now we are trying to increase $sp^3$ fraction of ta-C coating and control the coating parameters for thermal stability of thick ta-C at high temperatures.

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A facile synthesis of transfer-free graphene by Ni-C co-deposition

  • An, Sehoon;Lee, Geun-Hyuk;Jang, Seong Woo;Hwang, Sehoon;Yoon, Jung Hyeon;Lim, Sang-Ho;Han, Seunghee
    • Proceedings of the Korean Vacuum Society Conference
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    • 2016.02a
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    • pp.129-129
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    • 2016
  • Graphene, as a single layer of $sp^2$-bonded carbon atoms packed into a 2D honeycomb crystal lattice, has attracted much attention due to its outstanding properties. In order to synthesize high quality graphene, transition metals, such as nickel and copper, have been widely employed as catalysts, which needs transfer to desired substrates for various applications. However, the transfer steps are not only complicated but also inevitably induce defects, impurities, wrinkles, and cracks of graphene. Furthermore, the direct synthesis of graphene on dielectric surfaces has still been a premature field for practical applications. Therefore, cost effective and concise methods for transfer-free graphene are essentially required for commercialization. Here, we report a facile transfer-free graphene synthesis method through nickel and carbon co-deposited layer. In order to fabricate 100 nm thick NiC layer on the top of $SiO_2/Si$ substrates, DC reactive magnetron sputtering was performed at a gas pressure of 2 mTorr with various Ar : $CH_4$ gas flow ratio and the 200 W DC input power was applied to a Ni target at room temperature. Then, the sample was annealed under 200 sccm Ar flow and pressure of 1 Torr at $1000^{\circ}C$ for 4 min employing a rapid thermal annealing (RTA) equipment. During the RTA process, the carbon atoms diffused through the NiC layer and deposited on both sides of the NiC layer to form graphene upon cooling. The remained NiC layer was removed by using a 0.5 M $FeCl_3$ aqueous solution, and graphene was then directly obtained on $SiO_2/Si$ without any transfer process. In order to confirm the quality of resulted graphene layer, Raman spectroscopy was implemented. Raman mapping revealed that the resulted graphene was at high quality with low degree of $sp^3$-type structural defects. Additionally, sheet resistance and transmittance of the produced graphene were analyzed by a four-point probe method and UV-vis spectroscopy, respectively. This facile non-transfer process would consequently facilitate the future graphene research and industrial applications.

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Status of intestinal parasite infections in inhabitants of Palmar, Guavas Province, Ecuador (에꽈돌국 과야스주 빨말 본당 지역 주민의 장내 기생충 감염상태)

  • Jo, Seung-Yeol;Kim, Jung-Ho;Park, Seong-Hak
    • Parasites, Hosts and Diseases
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    • v.28 no.2
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    • pp.109-114
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    • 1990
  • Status of intestinal protozoan and helminthic infections was surveyed in mestizo population living in rural parish of Palmar and its nearby recintos(villages) , Three of the surveyed villages were at the Pacific coast and 5 villages were in inner pasture land, located about 100 km west of Guayaquil, the second largest city in Ecuador. One stool sample was examined by one cellophane thick smear for helminth ova and one direct smear stained with Lugol's iodine solution for protozoan cysts. Of 325 persons examined, 66.1% were positive for any ova or cyst. The positive rates were: 18.1% for Ascaris lumbricoides, 19.4% for Trichuris trichiura, 0.6% for hookworm, 3.7% for Hyntenolepis nana, 1.8% for Taenia sp., 19.4% for Entnmoeba histolytica, 28.6% for Entamoeba coli, 5.5% for Endolimax nasa, 1.5% for ledamoeba buetschlii, 11.l% for eiardia lamblia and 0.6% for Chilomastix mesnili. Poor supply of potable water was considered the main cause of high prevalence of intestinal protozoan infections.

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Studies on the promoted Aging of flue-cured leaf tobacco by cellulolytic Enzyme and Nicotinophiles (섬유소 분해효소 및 니코틴 분해세균을 이용한 잎담배의 발효촉진효과)

  • 이태호;성낙계
    • Journal of the Korean Society of Tobacco Science
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    • v.13 no.2
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    • pp.5-20
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    • 1991
  • For the quality enhancement of harvested-year leaf tobacco to the quality of 2-year naturally aged leaf tobacco, cellulose and nicotine degradative bacteria were isolated and identified. Effects of artificial fermentation treated cellulase and nicotine degradative bacteria on the quality of leaf tobacco were investigated from the chemical and sensory points of view. 1, Changes in chemical composition of leaf tobacco resulted from the addition of cellulase extracted from Cellulomonas sp. [3ml(${\mu}{\textrm}{m}$ D-glucose/ml. mil-1) of enzymes solution 11009 of leaf tobacco] and nicotine degradative bacteria, Pseudomonas sp. 2ml(IX109 cells$\div$ 100g of leaf tobacco), and subsequently fermented at 40${\mu}{\textrm}{m}$$^{\circ}C$, 65% R. H. for 40 days are as follows : 1) Content of crude fiber decreased 12% It took 9 min, 53 sec. to reach full combustion in control group but took only 7 min. 47 sec. in the treated group, taking almost equal time to 2-year naturally aged leaf tobacco(7 min. 35sec.). 2) Light intensity of control group was 60.96% with bright lemon color but that of treated leaf tobacco accounted for 47.69 with orange to dark brown color series, which was almost equal to the value, 45.69, of 2-year naturally aged leaf tobacco. 3) Linoleic acid, serving mild taste among organic acids, amounted to 1.llmg/g in control group but increased to 1.35m9/9 in the treated leaf tobacco, identical to the content(1.35mg/g) of 2-year naturally aged leaf tobacco. 4) Content of solanone, on of the typical leaf tobacco flavor compounds, accounted for 2.95% in control group but increased to 2.87% in treated group. 5) Methyl furan, useful flavor compound in smoke composition, accounted for 17.6$\mu\textrm{g}$/cig. in control group but increased to 25.9$\mu\textrm{g}$/cig. in treated group. However, acroleine decreased from 69.3$\mu\textrm{g}$/cig. in control group to 58.6$\mu\textrm{g}$/cig. in treated group 2. In sonsory test, mild taste evaluation of control group scored 5.47 and treated group 7.93 which was evaluted almost equal to the value(8.00) of 2-year naturally aged leaf tobacco. Aroma evaluation of control group scored 5.60, treated group 8.20, and 2-year naturally aged leaf tobacco 8.33. In addition, total harmony taste of control group showed 5.67, treated group 8.07 (p<0.01), and 2-year naturally aged leaf tobacco 8.00. From these results, it can be said that quality of treated leaf tobacco is not inferior to that 2-year naturally aged leaf tobacco.

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Studies on Screening and Iolation of ${\alpha}-Amylase$ Inhibitors of Soil Microorganisms( II ) -Isolation and Activities of the Inhibitor of Streptomyces Strain DMC-72- (토양균의 ${\alpha}-Amylase$ 저해제 검색 및 분리에 관한 연주(제2보) -스트렙토마이세스속 DMC-72 균주의 저해 성분의 분리 및 작용-)

  • Kim, Kyung-Jae;Lee, Shung-Hee;Kim, Jung-Woo;Kim, Ha-Won;Shim, Mi-Ja;Choi, Eung-Chil;Kim, Byong-Kak
    • The Korean Journal of Mycology
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    • v.13 no.4
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    • pp.203-212
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    • 1985
  • Of 450 strains isolated from the soil microbes collected in various locations in Korea, a strain had a strong inhibitory activity against bacterial ${\alpha}-amylase$ and was named strain DMC-72 of the genus Streptomyces. The amylase inhibitory metabolite produced by this strain was purified by means of acetone precipitation, adsorption on Amberlite IRC-50 and SP-Sephadex C-25. The inhibitor was found to be a derivative of oligosaccharides by spectral and chemical data. The inhibitor was stable at the pH range of $1{\sim}13$ and at $100^{\circ}C$ for half an hour, also inhibited other amylases such as salivary ${\alpha}-amylase$, pancreatic ${\alpha}-amylase$, fungal ${\alpha}-amylase$ and glucoamylase. However, it showed no inhibitory activity against ${\alpha}-glucosidase$, ${\beta}-glucosidase$, dextranase, and ${\beta}-amylase$. The kinetic studies of the inhibitor showed that its inhibitory effects on starch hydrolysis by ${\alpha}-amylase$ were noncompetitive.

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Enzymatic Properties of Pectin Esterase from Aspergillus sp. (Aspergillus속(屬)이 생성(生成)하는 Pectin Esterase의 효소학적(酵素學的) 성질(性質))

  • Song, Hyung-Ik;Yu, Tae-Shick;Chung, Ki-Taek
    • The Korean Journal of Mycology
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    • v.9 no.1
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    • pp.39-43
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    • 1981
  • The enzymatic properties of pectin esterase from Aspergillus sp. were studied. The optimum conditions for the enzyme reaction were pH 4.2 and temperature of $45^{\circ}C$. The crude enzyme was very stable at pH range of $2.2{\sim}4.6$, but about 20 percent of activity was lost at the range of pH $5.4{\sim}8.0$. The crude enzyme was very stable at $50^{\circ}C$ for one hour, however almost 100 percent of enzyme activity was lost at $70^{\circ}C$ for 30 minutes. The pectin esterase activity of crude enzyme was greatly inhibited by addition of sodium chloride at lower pH range. That is, the inhibition rates of enzyme activity at pH 3.0 and 4.0 were 47% and 28% in concentration of 1M sodium chloride, respectively. The enzyme activity was not affected by sodium chloride at pH 7.0 at different concentration of sodium chloride. Although the enzyme activity was not affected by addition of sucrose, it was slightly inhibited at higher concentration of sucrose.

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