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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.

Experimental Models of Schizophrenia (정신분열병의 실험적 모델)

  • Cheon, Jin-Sook
    • Korean Journal of Biological Psychiatry
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    • v.6 no.2
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    • pp.153-160
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    • 1999
  • Animal models can provide a useful tool for the study of some aspects of psychiatric disorders and their treatment. The four criteria for the evaluation of animal models of psychiatric disorders are as following : 1) similarity of inducing conditions 2) similarity of behavioral state 3) common underlying neurobiological mechanisms 4) reversal by clinically effective treatment techniques. Several animal models have been proposed for schizophrenia : phenylethylamine model, L-dopa model, hallucinogen model, cocaine model, amphetamine model, phencyclidine model, noradrenergic reward system lesion model, reticular stimulation model, social isolation model, conditioned avoidance reaction, catalepsy test, paw test, self-stimulation paradigms, latent inhibition paradigms, blocking paradigms, prepulse inhibition of the startle reflex, rodent interaction, social behavior in monkeys, hippocampal damage, high ambient pressure, and models using selective breeding. Among them, animals with bilateral lesion of the hippocampus may provide an adequate animal model for several symptoms of schizophrenia, and ketamine model can reproduce negative symptoms and cognitive deficits as well as positive symptoms of schizophrenia. In conclusion, no model of schizophrenia is entirely representative of the disease, and findings gleaned from model systems must be cautiously interpreted. Furthermore, the process of developing and validating animal models must work in concert with the process to identify reliable measures of human phenomenology.

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Bioactivities and Isolation of Functional Compounds from Decay-Resistant Hardwood Species (고내후성 활엽수종의 추출성분을 이용한 신기능성 물질의 분리 및 생리활성)

  • 배영수;이상용;오덕환;최돈하;김영균
    • Journal of Korea Foresty Energy
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    • v.19 no.2
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    • pp.93-101
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    • 2000
  • Wood of Robinia pseudoacacia and bark of Populus alba$\times$P. glandulosa, Fraxinus rhynchophylla and Ulmus davidiana var. japonica were collected and extracted with acetone-water(7:3, v/v) in glass jar to examine whether its bioactive compounds exist. The concentrated extracts were fractionated with hexane, chloroform, ethylacetate and water, and then freeze-dried for column chromatography and bioactive tests. The isolated compounds were sakuranetin-5-O-$\beta$-D-glucopyranoside from Populus alba $\times$Pl glandulosa, 4--ethyoxy-(+)-leucorobinetinidin frm R. pseudoacacia and fraxetion from F. rhynchophylla and were characterized by $^1H$ and$^{13}C $ NMR and positive FAB-MS. Decay-resistant activity was expressed by weight loss ratio and hyphae growth inhibition in the wood dust agar medium inoculated wood rot fungi. R. pseudoacacia showed best anti-decaying property in both test and its methanol untreated samples, indicating higher activity than methanol treated samples in hyphae grwoth test. In antioxidative test, $\alpha$-tocopherol, one of natural antioxidants, and BHT, one of synthetic antioxidants, were used as references to cmpare with the antioxidant activities of the extacted fractions. Ethylacetate fraction of F. rhynchophylla bark indicated the hightest activity in this test and all fractions of R. pseudiacacia extractives also indicated higher activities compared with the other fractions. In the isolated compounds, aesculetin isolated from F. rhynchophylla bark showed best activity and followed by robonetinidin from R. pseudoacaica.

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Isolation and Characterization of Starch-hydrolyzing Pseudoalteromonas sp. A-3 from the Coastal Sea Water of Daecheon, Republic of Korea (대한민국 대천 해안에서 분리한 전분 분해능을 갖는 Pseudoalteromonas sp. A-3 균주의 특징 및 동정)

  • Chi, Won-Jae;Park, Da-Yeon;Jeong, Sung-Cheol;Chang, Yong-Keun;Hong, Soon-Kwang
    • Microbiology and Biotechnology Letters
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    • v.39 no.4
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    • pp.317-323
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    • 2011
  • Strain A-3, an amylase-producing bacteria, was isolated from coastal seawater near Daecheon in the Republic of Korea. It was seen to possess a single polar flagella and grow well, on ASW-YP agar plates, at temperatures of between $20-37^{\circ}C$. However, it grew more slowly at the temperatures of $15^{\circ}C$ and $40^{\circ}C$. Similarly, it was observed to grow abundantly, in an Artificial Sea Water-Yeast extract-Peptone (ASW-YP) liquid medium, in a pH range of 6-9, but not grow at pHs of 4-5 and a pH of 10. Strain A-3 was noted as being close to Pseudoalteromonas phenolica O-$BC30^T$, Pseudoalteromonas luteoviolacea $NCIMB1893^T$, Pseudoalteromonas rubra $ATCC29570^T$, and Pseudoalteromonas byunsanensis $FR1199^T$, with 98.30%, 97.86%, 97.78%, and 97.25% similarities respectively, in its 16S rRNA sequence. A phylogenetic tree revealed that strain A-3 and P. phenolica O-$BC30^T$ belong to a clade. However, strain A-3 differed from P. phenolica O-$BC30^T$ in relation to a number of physiological characteristics. Strain A-3 exhibited no growth above 5% NaCl concentrations, no utilization of D-glucose, D-mannose, D-maltose, or D-melibose, and no lipase (C-14) activity. All of these properties strongly indicate that strain A-3 is distant from P. phenolica O-$BC30^T$ and thus led us to name it Pseudoalteromonas sp. A-3. Pseudoalteromonas sp. A-3 produces ${\alpha}$-amylase throughout growth. Maximal amylase activities of 144.48 U/mL and 149.20 U/mL were seen at pH 7.0 and $37^{\circ}C$, respectively. Pseudoalteromonas sp. A-3's high, stable production of ${\alpha}$-amylase in addition to its biochemical features, such as alkalitolerance, suggest that it is a good candidate for industrial applications.

Study on Low Temperature Tolerant Methane-Producing Bacteria for the Treatment of Agricultural and Livestock Wastes (농축산(農畜産) 폐기물(廢棄物) 처리(處理)를 위(爲)한 저온내성(低溫耐性) 메탄 생성균(生成菌)의 특성(特性)에 관(關)한 연구(硏究) 1. 저온조건(低溫條件)에서 시료별(試料別) 메탄 생성기작(生成機作) 연구(硏究))

  • Jung, Kwang-Yong;Kim, Jai-Joung
    • Korean Journal of Environmental Agriculture
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    • v.12 no.1
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    • pp.41-49
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    • 1993
  • The Study was conducted to develope the low temperature tolerant methane-producing bacteria(LTTB) and to increase the efficiency of anaerobic fermentation for the treatment of agricultural and livestock wastes at low temperature. The samples were collected from muddy soil, water logged sediment, organic layer and anaerobic sludge at three latitudes, $34.8{\sim}37.4\;^{\circ}N(Korea)$, $41.4\;^{\circ}N(USA)$ and $54.5{\sim}56.9\;^{\circ}N(Canada)$. They were used for determination of the methanogenesis rates for isolation and identification of the LTTB. The methanogenesis rate of smaples at low temperature were higher in the cellulose medium than methanol medium. The methanogenesis rate in the samples of subarctic region were $15{\sim}19$ moles/ml during 30 days at low temperature($8\;^{\circ}C$), whereas not detected in the samples of temperate region. The methanogenesis rate in the enrichment culture of subarctic samples were inhibited by the $40\;{\mu}g/ml$ of streptomycin + vancomycin or ampicillin + oleandomycin which were not effect to the methanogens. An inhabitation of high temperature tolerant methane producing bacteria was identified in the samples of temperate region, whereas that of the LTTB growing at $8{\sim}13^{\circ}C$ was identified in the subarctic region.

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Isolation and Characterization of Bacillus Species Having Antifungal Activity Against Pathogens of Ginseng Damping Off (인삼모잘록병원균에 항균활성을 갖는 Bacillus 균의 분리 및 특성조사)

  • Park, Kyeong Hun;Park, Hong Woo;Lee, Seong Woo;Lee, Seung Ho;Myung, Kyung Sun;Lee, Sang Yeob;Song, Jaekyeong;Kim, Young Tak;Park, Kyoung Soo;Kim, Young Ock
    • The Korean Journal of Pesticide Science
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    • v.20 no.4
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    • pp.380-387
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    • 2016
  • This study was performed to select potentially available biological control agent from soil bacteria for prevention of ginseng damping off. More than five hundred strains were isolated from ginseng rhizosphere soil. By testing antifungal activity, we have selected three soil bacteria strains and their ability to produce antibiotics and lytic enzymes such as cellulase, protease and pectate lyase was examined. Also, the presence of genes for biosynthesis of lipopeptide such as fengycin, bacillomycin D, surfactin, iturin A, and zwittermicin A was investigated in selected strains. All three strains produced cellulase, protease, and xylanase. Moreover, these strains had gene for biosynthesis of bacillomycin D, surfactin, and iturin A. ES1 and ES3 strains were identified Bacillus methylotrophucus and ES2 was confirmed Bacillus amyloliquefaciens using phylogenetic analysis on the basis of 16S rRNA gene sequences. In field test, control value of ES1, ES2 and ES3 treatment was 32.4%, 46.8% and 36.7%, respectively. This results indicate that antagonistic microbes with high ability of antifungal and lytic enzyme activity can be used as a useful biological control agent to control ginseng damping off.

Isolation, Identification, and Characteristics of Lactic Acid Bacteria for Production of Fermented Soymilk which Has Improved Sensory Quality (관능이 개선된 발효두유 제조를 위한 젖산균 분리·동정 및 특성)

  • Jung, Min-Gi;Kim, Su-In;Hur, Nam-Youn;Seong, Jong-Hwan;Lee, Young-Geun;Kim, Han-Soo;Chung, Hun-Sik;Kim, Dong-Seob
    • Microbiology and Biotechnology Letters
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    • v.44 no.1
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    • pp.74-83
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    • 2016
  • In order to improve the sour taste and foul odor of fermented soymilk, bacteria were isolated from kimchi and identified. Of the 89 bacterial strains isolated from kimchi, 3 isolates produced fermented soymilk with a sour taste and foul odor. The selected bacterial strains R53, R83, and R84 were identified by morphological, biochemical, and 16S rRNA analyses as Weissella koreensis. The strain R83, which produced fermented soymilk having the mildest sour taste and foul odor, was selected for further investigation and named W. koreensis KO3. The optimum culture condition for the fermentation of soymilk by W. koreensis KO3 was at $30^{\circ}C$ for 12 h. When soymilk was fermented under the optimum culture conditions, the viable cell count reached up to $8.71{\times}10^8CFU/ml$ and pH and acidity reached as low as 6.02 and as high as 0.33%, respectively. Twenty-seven amino acids and their derivatives were detected in fermented soymilk. The amounts of serine, glycine, threonine, alanine, and aspartic acid, which contribute to a sweeter taste, increased during fermentation. Orinithine, which was not detected before fermentation, increased during fermentation. Sensory evaluation showed that W. koreensis KO3-fermented soymilk has improved bean, roasted nut, and sour flavors as well as an enhanced mouthfeel, appearance, preferability, and overall acceptability compared with those of standard fermented soymilk. With further study and development, soymilk fermented by W. koreensis KO3 could serve as a health-promoting food with favorable sensory qualities.

Isolation and Chemical Analysis of Potent Anti-Complementary Polysaccharides from Fruiting Bodies of the Fomes fomentarius (말굽버섯 자실체에서 분리한 항보체 활성 다당체의 화학적 분석)

  • Park, Jung-Keun;Park, Kwe-Won;Shin, Kwang-Soon;Lee, Chang-Muk;Seok, Soon-Ja;Kim, Jeong-Bong;Koo, Bon-Sung;Han, Bum-Soo;Yoon, Sang-Hong
    • Microbiology and Biotechnology Letters
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    • v.41 no.2
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    • pp.198-206
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    • 2013
  • The five anti-complementary polysaccharides (MFKF-NP, MFKF-AP1${\alpha}$, ${\beta}$, and MFKF-AP2${\alpha}$, ${\beta}$) were separated from hot water extracts of fruiting bodies of Fomes fomentarius by two subsequent column chromatography using DEAE-sepharose FF and Concanavalin A-sepharose 4B. The order of anti-complementary activity was MFKF-AP1${\beta}$ > MFKF-AP1${\alpha}$ > MFKF-AP2${\alpha}$ > MFKF-AP2${\beta}$ > MFKF-NP > Polysaccharide Krestine (PSK). Especially, MFKF-AP1${\beta}$ among those showed the most excellent anti-complementary activity (70% of ITCH50 value at $20{\mu}g/ml$). The monosaccharide composition analysis by gas chromatography indicates that MFKF-AP1${\alpha}$ and ${\beta}$ are a kind of homoxylan consisted mainly of xylose above 97%. Molecular weight of MFKF-AP1${\beta}$, major anti-complementary polysaccharide, was estimated to be about 12,000 by high performance liquid chromatography (HPLC). After the incubation of the serum with MFKF-AP1${\beta}$ in the presence or absence of $Mg^{++}$ and $Ca^{++}$ ions, its anti-complementary activity was investigated. This result indicated that MFKF-AP1${\beta}$ seems to be activator both on the classical and the alternative pathway of complement activation.

Structural Identification of Antibiotics from Pseudomonas sp. RRj 228, a Antifungal Activity of Collectotrichum acutatum Causing Anthracnose on Pepper (Pseudomonas sp. RRj 228이 분비하는 항균물질의 동정과 고추탄저균 C. acutatum에 대한 항균활성)

  • Jeon, Sang-Yoon;Kim, Yong-Gyun;Lee, Sang-Mong;Son, Hong-Joo;Park, Hyean-Cheal;Kim, Sun-Tae;Park, Ki-Do;Kang, Ui-Gum;Kim, Keun-Ki
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1254-1260
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    • 2010
  • Microorganisms near the plant rhizosphere usually inhabit the surface or the inside of the plant roots and have a direct effect on plant growth by secreting plant growth promoters or antagonistic materials which protect the root zone system from various pathogens. This study was carried out to identify and isolate the antagonistic materials after isolation of microorganisms showing high antagonistic activities, in hopes of contributing to the development of sustainable agriculture and the preservation of agricultural environments. A number of antagonistic bacteria were isolated from paddy soil. Among isolates, RRj 228 showed plant growth promotion and antagonistic activity. RRj 228 was identified as Pseudomonas sp. according to the results of physiological properties and genetic methods. On the basis of the results of anti-fungal spectrum against several pathogens by RRj 228, the antagonistic effect of the isolate against Botrytis cinerea, Pythium ultimum, Phytopthola capsici, and Rhizoctonia solani, especially against red-pepper anthracnose caused by Colletotrichum acutatum, was remarkable. The experiment evaluating the biological control effect by RRj 228 revealed that the $ED_{50}$ value by the RRj 228 culture against C. acutatum, R. solani and P. ultimum were 0.14 mg/ml, 0.16 mg/ml and 0.29 mg/ml, respectively. An antagonistic substance was isolated and purified by several chromatographies from the RRj 228 culture. The $^1H$ and $^{13}C$ assignment of the antagonistic substance was achieved from two-dimensional $^1H-^1H$ COSY, HMQC, and HMBC. Finally, the antagonistic substance was identified as Phenazine-1-carboxylic acid ($C_{13}H_8N_2O_2$, M.W.=224).

Isolation of Strain for the Preparation of the Fermented Antler and Its Physiological Activities (발효녹용의 균주 선별 및 발효녹용의 생리활성)

  • Kim, Min-Kyung;Jung, Eun-Young;Lee, Hyun-Sun;Shin, Kwang-Soon;Kim, Yoo-Kyung;Ra, Kyung-Soo;Park, Chul-Soo;Woo, Moon-Jea;Lee, Sang-Hun;Kim, Jin-Soo;Suh, Hyung-Joo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.9
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    • pp.1237-1242
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    • 2009
  • This study was conducted to isolate strain for the preparation of fermented antler (Cervus cornu parvum) and evaluate its physiological activities. The growth degrees of twenty-one samples from Bacillus sp., Lactobacillius sp. and mushroom strain on antler extract agar were evaluated in this study, and Bacillus subtilis KH-15, SCB-3, Cordyceps militaris, Phellinus linteus, Inonotus obliquus 26136, and Inonotus obliquus 26147 were selected. The fermented antler extract by C. militaris had relatively higher contents of total sugar (1619.3 ${\mu}g$/mL), uronic acid (302.0 ${\mu}g$/mL), sulfated-glycosaminoglycan (S-GAGs) (119.9 ${\mu}g$/mL) and sialic acid (21.6 ${\mu}g$/mL) than any other extracts. The anti-complementary activities of all fermented antler extracts were higher than non-fermented antler extract, and among these samples, fermented antler extract by C. militaris showed the highest anti-complementary activity (inhibition of 50% total complement hemolysis, $ITCH_{50}$; 50.1% at 1,000 ${\mu}g$/mL). The ability of fermented antler extract by B. subtilis KH-15 to scavenge 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical ($IC_{50}$; 4.97 mg/mL) was significantly the highest (p<0.05), whereas the extract from I. obliquus exerted significantly (p<0.05) high 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity ($IC_{50}$; 16.98 mg/mL) among all samples. The results of this study suggest that physiological effects including immuno-modulating and antioxidant activities of the antler may be increased through fermentation process.