• Title/Summary/Keyword: 천연 추출물

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Isolation and Purification of Antimicrobial Peptide from Hard-shelled Mussel, Mytilus coruscus (참담치(Mytilus coruscus) 유래 항균 펩타이드 분리 및 정제)

  • Oh, Ryunkyoung;Lee, Min Jeong;Kim, Young-Ok;Nam, Bo-Hye;Kong, Hee Jeong;Kim, Joo-Won;An, Cheul Min;Kim, Dong-Gyun
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1259-1268
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    • 2016
  • In this study, we investigated antimicrobial peptide from the acidified muscle extract of Mytilus coruscus, which mostly inhabits China, Japan, and Korea, to develop a natural product-derived antibiotics substitution in terms of its abuse and restriction. Antimicrobial peptide was purified by $C_{18}$ reversed-phase high-performance liquid chromatography and was detected as having a molecular mass of 6,701 Da by MALDI-TOF/MS. The N-terminal amino acid sequence of the purified peak was obtained from edman degradation, and 20 identified residues shown 100% identity with the N-terminus region of sperm-specific protein and protamine-like PL-II/PL-IV precursor of Mytilus californianus. We also identified 60 open-reading frame (ORF) encoding amino acids with 183 bp of purified peptide based on the obtained amino acid residues. The amino acid sequence of ORF showed 100% and the nucleotide sequence revealed 97.2% identity with the protamine-like PL-II/PL-IV precursor of Mytilus californianus. Synthesized antimicrobial peptide showed antimicrobial activity against gram-positive bacteria, including Bacillus cereus (minimal effective concentration [MEC], $20.8{\mu}g/ml$), Bacillus subtilis (MEC, $0.2{\mu}g/ml$), Streptococcus mutans (MEC, $0.2{\mu}g/ml$), gram-negative bacteria including Pseudomonas aeruginosa (MEC, $5.7{\mu}g/ml$), Escherichia coli (MEC, $2.6{\mu}g/ml$) and fungi, Candida albicans (MEC, $56.3{\mu}g/ml$). In addition, synthesized peptide showed stable activities under heat and salt conditions against gram-positive and gram-negative bacteria, but was inhibited by salt against only C. albicans. With these results, isolated peptide from M. coruscus could be an alternative agent to antibiotics for defending against pathogenic microorganisms, and helpful information to understand the innate immune system of marine invertebrates.

Comparative Analysis of Technical System by Six Organic Rice Cultivation Type in the Southern Provinces (남부지방 벼 유기농법 유형별 투입기술체계 비교분석)

  • Ahn, In;Park, Joo-Seob;Kim, Sam-Hyun;Maeng, Woon-Young;Lee, In-Eae
    • Korean Journal of Organic Agriculture
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    • v.20 no.4
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    • pp.535-542
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    • 2012
  • This study was carried out from 2011 to 2012 in order to analyze the cultivation technology system by six organic rice cultivation types in the southern provinces for the purpose of utilizing organic rice farming guidelines. This surveys were conducted by site visit on 8 counties and 115 farms in southern area. Cultivation technology system by six organic rice cultivation types were cultivated by the principle of minimum input or non-input. cropping systems, a mixture of barley and rice, or rice green manure crops were grown. Cultivars were mixture of the local cultivars and Japanese varieties. Soil were managed by a self-manufactured compost, indigenous microorganisms and barley straw without any fertilization. Pest control was mainly practied by purchased eco-friendly organic materials or herbal medication residue and indigenous microbial self-manufactured. Weed was controled by snails and barley straw. In addition, considering the restore of a longtail pitching shrimp and grasshoppers, the fuction of environmental ecology of six organic rice cultivation type proved to be excellent all. Meanwhile, the productivity of the organic farming per 10a were the order of Life and environment > Natural circulation > Stevia > Ji jang > Art and nature > Taepyoung of Farming Methods, but the farmers received prices showed the opposite tendency. Simple profitablity did not show a big difference. But, the productivity of all type of the organic farming were lower then the conventional farming. Finally, If we want to conduct safety and easy farming, we can suggest stevia farming, Natural circulation farming and life and environmental farming out of 6 organic rice cultivation types.

Growth Inhibition Effect of Environment-friendly Farm Materials on Fungal Pathogens of Grape (친환경농자재의 포도 진균병 병원균에 대한 생장억제 효과)

  • Kim, Geon-Ju;Choi, Min-Kyung;Park, Jong-Han;Cha, Jae-Soon
    • Research in Plant Disease
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    • v.14 no.3
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    • pp.187-192
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    • 2008
  • Five environment-friendly farm materials including $Chitomate^{(R)}$, $Diegyun^{(R)}$, IC-$66D^{(R)}$, Gold $Bordo^{(R)}$, and $Biospot^{(R)}$ were examined for their growth inhibition effect of the 7 fungal pathogens of grape in vitro. $Diegyun^{(R)}$, being composed of natural ingredients which are extracted from a plant, was the most effective in suppression of mycelial growth of the fungi. $Diegyun^{(R)}$ inhibited the mycelial growth of all of fungi over 75% at $2,500{\mu}g{\cdot}mL^{-1}$ on potato dextrose agar(PDA) except Colletotrichum gloeosporioides 04-159. Growth inhibition effect of $Chitomate^{(R)}$, being composed of the chitosan, varied depending on the fungal pathogens on PDA. It inhibited the mycelial growth of the Botrytis cinerea 06-063 at the rate of 75.8% at $40,000{\mu}g{\cdot}mL^{-1}$ on PDA while it inhibited the mycelial growth of the C. gloeosporioides 04-159 at the rate of 6.5%. IC-$66D^{(R)}$ and Gold $Bordo^{(R)}$ are two different formula of the Bordeaux mixture, showed different control effects on mycelial growth inhibition. Except of Acremonium sp. the growth inhibition of IC-$66D^{(R)}$ was a little higher than Gold $Bordo^{(R)}$. $Biospot^{(R)}$, a chlorine formula, showed the strongest growth inhibition on C. gloeosporioides 04-159 among the farm materials used. Inhibition of spore germination of $Chitomate^{(R)}$, $Biospot^{(R)}$ and Gold $Bordo^{(R)}$ was higher than mycelial growth inhibition for Pseudocercospora vitis 04-152. The results suggest that the different types of environment-friendly farm materials are needed for different disease control in organic grape farm.

Safety of Nano-sized Bee Pollen in both In-vitro and In-vivo Models (생체 외 및 생체 내 실험조건에서 나노화 벌 화분의 안전성 규명)

  • Pyeon, Hae-In;So, Soojeong;Bak, Jia;Lee, Seunghyun;Lee, Seungmin;Suh, Hwa-Jin;Lim, Je-Oh;Kim, Jung-Woo;Kim, Sun Youn;Lee, Se Ra;Lee, Yong Hyun;Chung, Il Kyung;Choi, Yun-Sik
    • Journal of Life Science
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    • v.28 no.5
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    • pp.605-614
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    • 2018
  • Bee pollen has an outer wall which is resistant to both acidic and basic solutions and even the digestive enzymes in the gastrointestinal tract. Therefore, the oral bioavailability of bee pollen is only 10-15%. A previous study reported on wet-grinding technology which increased the extraction of active ingredients from bee pollen by 11 times. This study was designed to investigate the safety of wet-ground bee pollen. First, a single dose of wet-ground bee pollen was tested in both rats and beagle dogs at dosages of 5, 10, and 20 g/kg and 1.5, 3, and 6 g/kg, respectively. In rats, compound-colored stools were found in those administered 10 g/kg or more of wet-ground bee pollen. In beagle dogs, 6 g/kg of wet-ground bee pollen induced diarrhea in one male for four hours. However, no obvious clinical signs were found through the end of the experiment in rats and beagle dogs. In addition, no histological abnormality was found in all animals. The data indicates that a single dose of up to 20 g/kg of wet-ground bee pollen is safe. Next, the genetic toxicity of nano-sized bee pollen was tested. This study employed a bacterial reverse mutation test, a micronucleus assay, and a chromosomal aberration assay. In the micronucleus assay, there was no genetic toxicity up to the dosage of 2 g/kg. There was also no genetic toxicity in the bacterial reverse mutation test and chromosomal aberration assay. This data provides important information in developing nano-sized bee pollen into more advanced functional foods and herbal medicines.

Antioxidant Activities of Fractions Obtained from Dryopteris crassirhizoma, D. nipponensis and Polystichum lepidocaulon (관중, 참지네고사리 및 더부살이고사리 분획물의 항산화 활성)

  • Kim, Na-Rae;Kwon, Hyuk-Joon;Cho, Ju-Sung;Lee, Cheol-Hee
    • Korean Journal of Plant Resources
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    • v.25 no.2
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    • pp.176-183
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    • 2012
  • This study was carried out to develop ferns as the natural antioxidant materials by graduating and extracting fronds of $Dryopteris$ $crassirhizoma$, $Dryopteris$ $nipponensis$, and $Polystichum$ $lepidocaulon$, which belong to Dryopteridaceae, using solvent, and analyzing the antioxidant effect of each fraction. The n-butanol fraction of $D.$ $crassirhizoma$ (550.0 $mg{\cdot}g^{-1}$), the ethyl acetate fraction of $D.$ $nipponensis$ (374.8 $mg{\cdot}g^{-1}$), and the n-butanol fraction of $P.$ $lepidocaulon$ (781.8 $mg{\cdot}g^{-1}$) showed relatively higher total contents of polyphenol. The chloroform fraction of $D.$ $crassirhizoma$ (72.9 $mg{\cdot}g^{-1}$), and the n-hexane fraction of $D.$ $nipponensis$ (72.9 $mg{\cdot}g^{-1}$) and $P.$ $lepidocaulon$ (154.5 $mg{\cdot}g^{-1}$) contained relatively higher total contents of flavonoids. DPPH radical scavenging activity was most excellent in the n-butanol fraction of $D.$ $crassirhizoma$ ($RC_{50}=0.02mg{\cdot}mL^{-1}$) and $P.$ $lepidocaulon$ ($RC_{50}=0.04mg{\cdot}mL^{-1}$), and the water fractions of $D.$ $nipponensis$ ($RC_{50}=0.01mg{\cdot}mL^{-1}$). ABTS radical scavenging activity was potent in the n-hexane and n-butanol fractions of $D.$ $crassirhizoma$ (each $RC_{50}=0.02mg{\cdot}mL^{-1}$), the ethyl acetate fraction of $D.$ $nipponensis$ ($RC_{50}=0.03mg{\cdot}mL^{-1}$), and the n-butanol fraction of $P.$ $lepidocaulon$ ($RC_{50}=0.06mg{\cdot}mL^{-1}$). There was the large amount of total polyphenol content in the n-butanol fraction of $D.$ $crassirhizoma$ and $P.$ $lepidocaulon$, and their radical scavenging activities were potent. Therefore, it was thought that biologically active substances of each fraction layer are required to be analyzed and used.

High Yield Bacterial Expression and Purification of Active Cytochrome P450 p-coumarate-3-hydroxylase (C3H), the Arabidopsis Membrane Protein (대장균 시스템을 이용한 Arabidopsis 막 단백질 cytochrome P450 p-coumarate-3hydroxylase (C3H) 활성형의 과발현 및 분리정제)

  • Yang, Hee-Jung;Kim, Wan-Yeon;Yun, Young-Ju;Yoon, Ji-Won;Kwon, Tae-Woo;Youn, Hye-Sook;Youn, Bu-Hyun
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1039-1046
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    • 2009
  • The cytochrome P450s (P450s) metabolizing natural products are among the most versatile biological catalysts known in plants, but knowledge of the structural basis for their broad substrate specificity has been limited. The activity of p-coumarate 3-hydroxylase (C3H) is thought to be essential for the biosynthesis of lignin and many other phenylpropanoid pathway products in plants however, all attempts to express and purify the protein corresponding C3H gene have failed. As a result, no conditions suitable for the unambiguous assay of the enzyme are known. The detailed understanding of the mechanism and substrate-specificity of C3Hdemands a method for the production of active protein on the milligram scale. We have developed a bacterial expression and purification system for the plant C3H, which allows for the quick expression and purification of active wild-type C3H via introduction of combinational mutagenesis. The modified cytochrome P450 C3H ($C3H_{mod}$) could be purified in the absence of detergent using immobilized metal affinity chromatography and size exclusion chromatography following extraction from isolated membranes in a high salt buffer and catalytically activated. This method makes the use of isotopic labeling of C3H for NMRstudies and X-ray crystallography practical, and is also applicable to other plant cytochrome P450 proteins.

Free Amino Acid and Polysaccharide Content of Submerged Mycelial Culture of Fomitopsis pinicola (귤피 및 녹차추출물에서 배양한 소나무잔나비버섯(Fomitopsis pinicola) 균사체의 유리아미노산 및 다당류 함량)

  • Jang Kyung-Ho;Shin Kyung-Ok;Kim Soon-Dong
    • Food Science and Preservation
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    • v.12 no.4
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    • pp.379-386
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    • 2005
  • Free amino acids and polysaccharide content of submerged mycelial culture of Fomitopsis pinicola using $2\%$ citrus peel water extracts(CP), $2\%$ green tea water extracts(GT) and yeast-malt broth(YM) were investigated. Cultivations were conducted at $30^{\circ}C$ and 150 rpm for 10 days. Yields of the mycelium as fresh weight basis in GT-, CP- and YM-broth were $42.3\%,\;34.2\%\;and\;9.89\%$, and their turbidity(OD at 660 nm) of the broth without mycelium were 0.14, 0.16 and 0.22, respectively. Total free amino acid content in the mycelium were 928.19 $mg\%$ in YM, 1060.53 $mg\%$ in CP, 764.83 $mg\%$ in GT, and the major free amino acid was lysine in YM, glutamic acid in CP and GT. Total free exo-amino acid contents were 659.75 $mg\%$ in YM, 954.55 $mg\%$ in CP, 838.69 $mg\%$ in GT, and the major in the all broths was glutamic acid Total amino acid derivatives content of the mycelium were in order of CP>GT>YM, and the major was cystathionine in YM, hydroxy proline in CP and GT. The major among exo-amino derivatives was hydroxy proline in the all broth. Exo-AIS content was in order of CP>GT>YM. Acid soluble polysaccharide content of the mycelium was GT($0.69\%$)>YM($0.39\%$)>CP($0.18\%$). The exo-polysaccharide content was in order of GT($0.87\%$)>CP($0.69\%$)>YM($0.09\%$). Alkali soluble polysaccharide content of the mycelium was in order of CP($5.21\%$)>GT($5.18\%$)>YM($4.56\%$), and exo-polysaccharide was in order of GT($6.79\%$)>YM($3.57\%$)>CP($3.01\%$). The alkali soluble polysaccharide eluted from mycelium cultivated in CP broth was supposed to polysaccharide(about 500,000 daltons) composed of hexose and uronic acid bounded with protein(below 10,000 daltons).