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Development of TaqMan probe-based real-time PCR for rapid identification of beef, pork and poultry meat (소, 돼지, 가금육류의 신속한 동정을 위한 TaqMan probe를 이용한 real-time PCR 개발)

  • Koh, Ba-Ra-Da;Kim, Ji-Yeon;Na, Ho-Myung;Park, Seong-Do;Kim, Yong-Hwan
    • Korean Journal of Veterinary Service
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    • v.35 no.3
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    • pp.215-222
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    • 2012
  • Species-specific $TaqMan^{(R)}$ probe-based real-time PCR assays were developed for detection of beef, pork, chicken, duck, goose and turkey. The primer and probe sets used in this study were designed to be complementary to fibroblast growth factor (FGF) for cattle and pig, mitochondrial NADH dehydrogenase (ND) subunit 3 and ND2 for chicken and duck, 12S rRNA for goose and turkey, respectively. As internal positive control we used conserved region in the ribosomal 18S RNA gene to ensure the accuracy of the detection of target DNA by real-time PCR. We confirmed that real-time PCR assays with the primer and probe sets were positive for cattle, pig and chicken intended target animal species with no cross-reactivity with other non-target animal species. Only >50 ng DNA of beef show cross-reactivity in the determination of duck. Using species-specific primer and probe sets, it was possible to detect amounts of 0.1 ng DNA of cattle and pig, 1.0 pg DNA of chicken, duck and turkey, and 0.1 pg DNA of goose for raw samples, respectively. The detection limits were 0.1 ng DNA of cattle, 1.0 ng DNA of pig and 1.0 pg DNA of chicken for DNA mixtures (beef, pork and chicken) extracted from heat-treated ($121^{\circ}C$/5 min) meat samples. In conclusion, it can be suggested that the $TaqMan^{(R)}$ probe-based assay developed in this study might be a rapid and specific method for the identification of meat species in raw or cooked meat products.

Antioxidant, Antidiabetic, and Anti-Inflammatory Effects of Extracts and Fractions from Parthenocissus tricuspidata Stems (담쟁이덩굴 추출물과 분획물의 항산화, 항당뇨 및 항염증 효과)

  • Cho, Eun Kyung;Choi, Young Ju
    • Journal of Life Science
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    • v.23 no.3
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    • pp.399-405
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    • 2013
  • This study was carried out to analyze the effects on antioxidative, antidiabetic, and anti-inflammatory activities of Parthenocissus tricuspidata (PT) stem extracts. The total phenolic contents of hot water and ethanol extracts from PT stems were 61.5 mg TAE/g and 122.1 mg TAE/g, respectively. The antioxidative activities of hot water and ethanol extracts from PT stem were measured by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and superoxide dismutase (SOD) assay. The DPPH radical scavenging activities of ethanol extract and butanol fraction were approximately 95% and 92% at 1 mg/ml, respectively, and the SOD activities of ethanol extract and butanol fraction were about 91% and 97% at 1 mg/ml, respectively. The DPPH radical scavenging and SOD activities of ethanol extract and butanol fraction from PT stem increased remarkably increased in a dose-dependent manner and were higher than in the hot water extracts. Compared to the acarbose, a known anti-diabetic drug, which was used as a positive control, the ${\alpha}$-glucosidase inhibitory capacity of PT stem showed a strong inhibitory rate in ethanol extract and in butanol and hexane fractions. We investigated the effect of hot water extract from PT stem on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 cells. Hot water extract from PT stem inhibited LPS-induced NO production up to 40% at a treatment of 1 mg/ml. These results suggest that PT stem extracts have an effect on antioxidative, antidiabetic, and anti-inflammatory activities and thus have great potential as antidiabetic materials and a source for natural health products.

Novel Alkali-Tolerant GH10 Endo-${\beta}$-1,4-Xylanase with Broad Substrate Specificity from Microbacterium trichothecenolyticum HY-17, a Gut Bacterium of the Mole Cricket Gryllotalpa orientalis

  • Kim, Do Young;Shin, Dong-Ha;Jung, Sora;Kim, Hyangmi;Lee, Jong Suk;Cho, Han-Young;Bae, Kyung Sook;Sung, Chang-Keun;Rhee, Young Ha;Son, Kwang-Hee;Park, Ho-Yong
    • Journal of Microbiology and Biotechnology
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    • v.24 no.7
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    • pp.943-953
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    • 2014
  • The XylH gene (1,167-bp) encoding a novel hemicellulase (41,584 Da) was identified from the genome of Microbacterium trichothecenolyticum HY-17, a gastrointestinal bacterium of Gryllotalpa orientalis. The enzyme consisted of a single catalytic domain, which is 74% identical to that of an endo-${\beta}$-1,4-xylanase (GH10) from Isoptericola variabilis 225. Unlike other endo-${\beta}$-1,4-xylanases from invertebrate-symbiotic bacteria, rXylH was an alkali-tolerant multifunctional enzyme possessing endo-${\beta}$-1,4-xylanase activity together with ${\beta}$-1,3/${\beta}$-1,4-glucanase activity, which exhibited its highest xylanolytic activity at pH 9.0 and 60oC, and was relatively stable within a broad pH range of 5.0-10.0. The susceptibilities of different xylosebased polysaccharides to the XylH were assessed to be as follows: oat spelts xylan > beechwood xylan > birchwood xylan > wheat arabinoxylan. rXylH was also able to readily cleave p-nitrophenyl (pNP) cellobioside and pNP-xylopyranoside, but did not hydrolyze other pNP-sugar derivatives, xylobiose, or hexose-based materials. Enzymatic hydrolysis of birchwood xylan resulted in the product composition of xylobiose (71.2%) and xylotriose (28.8%) as end products.

VOC/HAPs Emission Characteristics & Adsorption Evaluation for Paint Products in Busan Area (부산지역 페인트제조시설의 VOC/HAPs 배출특성 및 흡착능 평가)

  • Song, Bok-Joo;Lee, Seung-Min;Cho, Gab-Je;Cho, Jeong-Gu;You, Pyung-Jong;Kim, Gi-Gon
    • Journal of Korean Society of Environmental Engineers
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    • v.34 no.5
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    • pp.316-325
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    • 2012
  • In this study, we suggested adequate control technology by analyzing emission process and main chemical of VOC/HAPs generated from four paint factories in Busan area. And we estimated whether Kapok fiber, which is a natural material, can be commercially used for an absorbent by testing adsorption ability. As a result of this sturdy, above 60% of VOC/HAPs was volatilized and dispersed inside the working place without conducting control system of facilities during manufacturing paints. Concentration profile of VOC/HAPs, which is volatile naturally outside the factories, is surveyed above 70% at Toluene, Ethylbenzene, and Xylene. And a result of evaluation of odor attribution level about the component whose Odor Threshold is known, it is estimated that major cause material of A, B, C factories is Toluene and that of D factory is m/p-Xylene. And that result presented design arguments such as facilities specifications, activated carbon filling volume, and replace cycle of activated carbon as control technology. Also, that result presented emission process improvement such as adsorption of central-controlled ventilation device, installation of inlet flenge, and potable cleaning process. The rate of pollutant adsorption of Kapok fiber, which is natural material, is indicated about 91.9%, 66.7%. That result validated the possibility as replacement of activated carbon.

한국산 장뇌산삼의 부위별 유용성분함량 및 추출용매조건의 영향

  • 김준한;문혜경;강우원;김종국
    • Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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    • 2003.10a
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    • pp.150.2-151
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    • 2003
  • 한국산 장뇌산삼의 열매, 잎, 줄기 및 뿌리를 -7$0^{\circ}C$ 동결건조 분말화시킨 시료에 대한 추출용매와 농도에 따른 유용성분의 함량을 비교, 분석하였다. 추출액의 당도는 잎과 줄기의 80% 에탄올추출액이 각각 22.58%와 22.53%로 가장 높았고, pH는 4.43-7.41 수준이었고, 갈변도는 흡광도가 잎과 뿌리 80% 에탄올추출액이 1.803과 1.085로 가장 높았다. 색도의 경우 L값는 줄기 100% 증류수추출액이 24.56, 열매 80% 메탄올추출액이 24.35로 가장 높았고, 뿌리와 잎 80% 에탄올추출액이 각각 17.47과 19.59로 가장 낮은 값을 나타내었다. a값은 잎100% 증류수추출액이 0.41로 가장 높았고, 줄기 80%메탄올추출 액이 -0.49로 가장 낮은 값을 보였다. b값은 줄기 80% 메탄올추출액이 3.69로 가장 높았고, 열매 100% 증류수추출액 이 0.45로 가장 낮은 값이었다. 주요 유리당은 sucrose, glucose 및 fructose 이었고, 유리당 총함량은 열매와 줄기 100% 증류수추출액이 각각 6733 mg/100g과 6142 mg/100g으로 가장 많은 량을 함유하였고, sucrose는 뿌리 80% 메탄올추출액 이 3673 mg/100g으로, glucose 및 fructose는 줄기 80% 에탄올추출액과 잎 80% 메탄올추출액에 각각 4283 mg/100g과 1897 mg/100g으로 높은 함유량을 나타내었다. 또한 잎과 줄기에는 xylose가 304-524 mg/100g 수준으로 함유되어 있었고, 뿌리에는 maltose가 소량 함유하고 있었다. 주요 유기산으로는 citric acid, tartaric acid 및 malic acid가 확인되었고, citric acid는 뿌리 80% 메탄올추출액이 1849 mg/100g으로 가장 높은 함량이었고, tartaric acid는 잎 80% 메탄올추출액이 3263 mg/100g으로 가장 높은 함량이었고, malic acid는 뿌리 80% 메탄올추출액이 1856 mg/100g으로 가장 높은 함량이었으며, 뿌리 80% 메탄올추출액에는 succinic acid, malonic acid 및 oxalic acid 등이 확인되었다. 유리아미노산은 L-Arginine, ${\gamma}$-Amino-n-butyric acid, Ethanolamine, L-Proline, $\beta$-Alanine 및 L-sarcosine 등 총 35종이 확인되었고, L-Arginine은 251-7379 mg/100g 수준으로 특히, 뿌리 80% 메탄올추출액에는 1319 mg/100g으로 총함유량의 79.13%로 매우 높은 함유량을 나타내었다. P, K, Na, Ca 및 Mg 등이 주된 무기질로 확인되었고, 그 중 P는 줄기 100% 증류수추출액에 15563 mg/100g으로 가장 높았고, K은 잎 80% 메탄을 추출액에 4952 mg/100g, Ca과 Na은 잎과 열매 100% 증류수추출액에 각각 3052 mg/100g과 1798 mg/100g, Mg은 잎 100% 증류수추출액에 950 mg/100g으로 매우 높은 함유량을 보였고, 또한 미량원소로는 Zn, Cu, Cr, Mn, Co, Mo, Fe 등이 함유되어 있었다.

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Development of the Formula for Natural Bread-making Starter (천연제빵 발효 Starter의 개발)

  • 이종열;이시경;조남지;박원종
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.8
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    • pp.1245-1252
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    • 2003
  • A starter formulation was developed to substitute a baker's yeast with natural starter when manufacturing bread products. To develop an active starlet, starter was formulated varying with types of wheat flours, level of water contents and various nutrients. Activities of starter were investigated in terms of viable counts of microbes and change of pH and total titratible acidity Domestic wheat flours contain 100 times more number of lactic acid bacteria than yeast regardless of types of wheat flours. The more protein contents in wheat flours, the more stable microbes in starter. This was considered to be the result of buffering effect of wheat proteins. The optimum level of protein content to ensure the activity of starter was more than 12.0%. Optimum level of water content in active starter was 110% based on strong flour. The more water or the less water had the tendency of decreasing viable counts of microbes. Addition of salt and sucrose had increased the activity of starters. However oligosaccharides did not affect the activity of starter. The optimum concentrations of salt and sucrose were 1.0% and 5.0% respectively. Bread with the starter was higher scored than breads with yeast in terms of all the quality and sensory characteristics except their volumes. In conclusion, a starter formulated with strong flour 100%, water 110%, salt 1% and sucrose 5% was considered to have high potential as a substitute of yeast in making natural bread.

Mechanism of Melanogenesis Inhibition by Melanoston Isolated from Yeast (효모에서 분리한 멜라닌 생성 억제 물질의 작용 기전)

  • Lee, Seung-Sun;Jung, Ho-Kwon;Oh, Chul;Choi, Tae-Boo
    • KSBB Journal
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    • v.19 no.2
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    • pp.118-124
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    • 2004
  • Melanocytes synthesize melanin within discrete organelle termed melanosomes which are transferred to the surrounding keratinocytes and can be produced in varying sizes, numbers and densities. Skin whitening products have become increasingly popular in the past few years. The most successful natural skin whitening agents are: Arbutin, Vitamin C, Kojic acid, Mulberry, which are all tyrosinase inhibitors. In this work, melanoston, a melanogenesis inhibitor isolated from yeast was studied to understand its mechanism of melanogenesis inhibition. It was found that melanoston was not a tyrosinase inhibitor, while when melanoston was applied to the B16 melanoma cell culture media, the intracellular tyrosinase activity was decreased by more than 30%, When B16 melanoma was stimulated with ${\alpha}$-MSH, cell morphololgy was dramatically changed to have lots of dendrites on the cell membrane surface. On the other hand, B16 was treated with ${\alpha}$-MSH and melanoston, simultaneously, the change of cell morphology was not so great. This inhibition effect of melanoston was found to be related to the inhibition of intracellular activation and transportation of tyrosinase, which was observed by immunostaining of B16 melanoma using anti-tyrosinase antibody. From these results, melanoston was regarded as an inhibitor to the differentiation of melanoma cells.

Structural Characteristics of Expression Module of Unidentified Genes from Metagenome (메타게놈 유래 미규명 유전자의 발현에 관련된 특성분석)

  • Park, Seung-Hye;Jeong, Young-Su;Kim, Won-Ho;Kim, Geun-Joong;Hur, Byung-Ki
    • KSBB Journal
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    • v.21 no.2
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    • pp.144-150
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    • 2006
  • The exploitation of metagenome, the access to the natural extant of enormous potential resources, is the way for elucidating the functions of organism in environmental communities, for genomic analyses of uncultured microorganism, and also for the recovery of entirely novel natural products from microbial communities. The major breakthrough in metagenomics is opened by the construction of libraries with total DNAs directly isolated from environmental samples and screening of these libraries by activity and sequence-based approaches. Screening with activity-based approach is presumed as a plausible route for finding new catabolic genes under designed conditions without any prior sequence information. The main limitation of these approaches, however, is the very low positive hits in a single round of screening because transcription, translation and appropriate folding are not always possible in E. coli, a typical surrogate host. Thus, to obtain information about these obstacles, we studied the genetic organization of individual URF's(unidentified open reading frame from metagenome sequenced and deposited in GenBank), especially on the expression factors such as codon usage, promoter region and ribosome binding site(rbs), based on DNA sequence analyses using bioinformatics tools. And then we also investigated the above-mentioned properties for 4100 ORFs(Open Reading Frames) of E. coli K-12 generally used as a host cell for the screening of noble genes from metagenome. Finally, we analyzed the differences between the properties of URFs of metagenome and ORFs of E. coli. Information derived from these comparative metagenomic analyses can provide some specific features or environmental blueprint available to screen a novel biocatalyst efficiently.

Selection of Mutant Silkworm with Oxidation-deficient Haemolymph for Insect Cell Culture (곤충세포 배지 개발을 위한 체액산화지연 돌연변이 누에계통 선발)

  • Choi, Ji-Young;Kim, Jong-Gill;Choi, Young-Cheol;Yoon, Hyung-Joo;Ahn, Mi-Young;Kim, Sam-Eun;Hwang, Seok-Jo
    • Journal of Sericultural and Entomological Science
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    • v.49 no.2
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    • pp.47-50
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    • 2007
  • Insect cell culture system has been demonstrated the effective means of producing medical and agricultural products. Furthermore, Fetal bovine serum (FBS) is in wide use in insect cell culture. Silkworm hemolymph was tested to develop as a substitute for FBS and was effective in insect cell growth. Hemolymph is oxidized and darkens visibly during the collection from silkworms due to the activity of tyrosinase in it. Toxic quinones are produced by the oxidation and consequently inhibit the cell growth. Heat treatment can be used to prevent the oxidation; however, the oxidation may occur during the collection of hemolymph before it is heat-treated. Hemolymphs collected from 257 different strains of silkworms were examined to select the slowly oxidized hemolymphs. Hemolymphs collected from mutant strains such as $Y_4$, TBO and $wE^b$ showed relatively slow color changes. Oxidation rates of the hemolymphs were measured by the absorbance change using a spectrophotometer. The absorbance of mutant hemolymph reached the saturation value at $20^{\circ}C$ in each 330 min ($Y_4$), 360 min (TBO) and 450 min ($wE^b$) min, whereas the total oxidation time of the wild-type (Baekokjam) hemolymph at the same temperature was 120 min. The cell growth in the medium supplemented with mutant species hemolmph was more effective that in the medium supplemented with Baekokjam species hemolymph.

Pigment Distribution Analysis of High Speed Fan for Dusting Large Pasture at Livestock Farms (축산농가를 위한 대규모 목초지 방제용 고속 팬의 실험을 통한 색소 분포에 따른 분석)

  • Kim, C.S.;Min, B.R.;Seo, K.W.;Lee, D.W.
    • Journal of Animal Environmental Science
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    • v.15 no.2
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    • pp.99-106
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    • 2009
  • We conducted a comparison experiment of our wide-area spraying high speed fan with a fan which was developed by Japan MARUYAMA Inc. have been much recognized for it's history and skills in a field of spraying machine. In result, MARUYAMA Inc. fan sprayed up to 120 m and, on the other hand, wide-area spraying high speed fan of our own making was able to spray up to more than 160 m. Wide-area spraying machine have been supplied to domestic demands by importing from Japan, but it is determined to be possible that home products will occupy market for it's ability, price and A/S environment in future. The main research results are below. Some plastic cups which are in the open market are used for our experiment. After distributing 90 cups in a range of 10m vertically with localizing 5 cups horizontally and 18 cups vertically, spraying machine was moved and finally we make distribution charts by estimating the sprayed amounts on each cup. Vertical distance was set up to 5m and we calculated average values by using sprayed amount and data of 4 observers. In result our fan showed much sprayed amounts than MARUYAMA Inc. all over the field except situations of vehicle departing and stopping.

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