• Title/Summary/Keyword: species characterization

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Purification of an Antibacterial Peptide from the Gills of the Pufferfish Takifugu pardalis (졸복의 아가미로부터 항균성 펩타이드의 정제)

  • Kim, Tae Young;Go, Hye-Jin;Park, Nam Gyu
    • Journal of Life Science
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    • v.27 no.1
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    • pp.50-56
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    • 2017
  • An antibacterial peptide was purified from an acidified gill extract of the pufferfish Takifugu pardalis. The acidified gill extract was put through a Sep-Pak C18 solid phase extraction cartridge using a stepwise gradient and divided into a flow-through (F.T.) and 60% methanol fraction (RM 60). Among the eluents, RM 60 had potent antibacterial activity against Bacillus subtilis KCTC 1021. RM 60 was partially purified on a cationic-exchange column (SP-5PW) by a linear gradient, and the antibacterial peptide was then further purified, using a series of cationic-exchange and $C_{18}$ reversed-phase HPLC columns. For characterization of the purified peptide, its molecular weight and amino acid sequence were analyzed by MALDI-TOF MS and Edman degradation. The molecular weight of the peptide was about 1171.6 Da. The amino acid sequence of the peptide was partially determined as: STKEKAPRKQ. A comparison of the N-terminal amino acid sequence of the purified peptide with that of other known polypeptides revealed high homology with the N-terminus of the histone H3 protein, which belongs to the histone H3 family. Thus, this peptide was designated as a puffer fish gill (PFG)-related antimicrobial peptide. This is the report to describe an antimicrobial function for the N-terminus of histone H3 of an animal species. The findings suggest that this peptide plays a significant role in the innate defense system of the pufferfish.

Preparation and Characterization of Phytochemical-Rich Extract from Sasa quelpaertensis Leaf (식물화합물 다량 함유 제주조릿대 잎 추출물의 제조와 특성)

  • Lee, Ju Yeop;Ko, Hee Chul;Jang, Mi Gyeong;Kim, Se Jae
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1330-1335
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    • 2016
  • Sasa species leaves have been used in traditional medicine for their anti-inflammatory, antipyretic, and diuretic properties. Sasa quelpaertensis Nakai is a small bamboo grass that grows only on Mt. Halla on Jeju Island, Republic of Korea. This small bamboo grass has recently been the focus of much attention due to its potential biomass as well as its beneficial health effects. In this study, to promote the efficient utilization of the S. quelpaertensis leaf, we established a simple preparation method for phytochemical-rich extract (PRE) by comparing phytochemical contents and biological activities according to extraction methods. high performance liquid chromatography (HPLC) analysis revealed that the contents of two major phytochemicals such as, tricin (5.35 mg/g) and p-coumaric acid (44.10 mg/g) contained in PRE were higher than those in fresh hot water extract (SQH, p-coumaric 23.39 mg/g, tricin 0.18 mg/g) and ethanol extract (SQE, p-coumaric 10.8 mg/g, tricin 0.38 mg/g). The antioxidant activities [1,1-diphenyl-2-picrylhydrazy (DPPH) radical scavenging activity, 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activity, nitric oxide (NO) scavenging activity, and xanthine oxidase inhibitory activity] of PRE were higher than those of SQH and SQE. PRE effectively inhibited NO production in LSP- stimulation RAW 264.7 cells, and the growth of human promyelocytic leukemia (HL-60) cells. These results suggest that PRE has a potential as a promising antioxidant and anti-inflammatory agent.

Molecular Cloning and mRNA Expression of the Bovine Peroxisome Proliperator Receptor Gamma(PPARγ) (한우 PPARγ 유전자의 동정과 mRNA의 발현)

  • Jeoung, Y.H.;Lee, S.M.;Park, H.Y.;Yoon, D.H.;Choi, J.G.;Moon, S.J.;Kang, M.J.
    • Journal of Animal Science and Technology
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    • v.46 no.1
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    • pp.23-30
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    • 2004
  • The peroxisome proliferator-activated receptor $\gamma$(PPAR$\gamma$), a member of the steroid/thyroid nuclear hormone receptor suferfamily of ligand-activated transcription factor, is an important regulator of adipocyte gene expression and differentiation. In this studies, we report the identification, characterization, and expression of a Hanwoo PPAR$\gamma$ gene. The PPAR$\gamma$ cDNA sequence of the Hanwoo show strong conservation with the corresponding sequences reported in other species except of three amino acid sequences. The distribution of PPAR$\gamma$ mRNA in various tissues of Korean cattle aged 12 months were investigated using Northern Blot analysis. The highest expression was detected in adipose tissue, more lower expression was detected in colon, small intestine, kidney, lung, while expression was not detected in brain, heart. PPAR$\gamma$ expression was higher in adipose tissue of Korean cattle when aged 30 months than aged 12 months. These results indicated PPAR$\gamma$, regulator adipocyte gene expression and differentiation, related on adipose differentiation in Korean native cattle(HANWOO).

Isolation and Partial Characterization of Isoflavone Transforming Lactobacillus plantarum YS712 for Potential Probiotic Use (Isoflavone 비배당화 및 항산화 활성을 지닌 Lactobacillus plantarum YS712의 선발)

  • Cho, Yoon-Hee;Imm, Jee-Young;Kim, Hwa-Young;Hong, Seong-Gil;Hwang, Sung-Joo;Park, Dong-Jun;Oh, Se-Jong
    • Food Science of Animal Resources
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    • v.29 no.5
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    • pp.640-646
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    • 2009
  • Lactic acid bacteria (LAB) are typical probiotic microbes that are used in various industries including fermented foods, feed additives, and pharmaceuticals. The purpose of this study was to compare the ability of isoflavone biotransformation and antioxidative activity of 17 LAB. Six strains including the Lactobacillus species exhibited a 100% hydrolysis rate for daidzein during fermentation. The content of total genistein in soymilk fermented with these strains was $872-943\;{\mu}g/g$. The DPPH (1, 1-diphenyl-2-picrylhydrazyl) radical scavenging ability of the LAB was widely variable and ranged from 23-78%. A selected strain was isolated from kimchi and the strain was identified as Lactobacillus plantarum ssp. through the API carbohydrate fermentation pattern and 16S rDNA profile. The strain exhibited excellent acid tolerance in an artificial gastric solution. L. plantarum YS712 showed high $\beta$-glucosidase activity in fermentation. The concentration of genistein and daidzein in soymilk fermented with L. plantarum YS712 increased from 3.64 to $917.3\;{\mu}g/g$ and from 58.18 to $1062.17\;{\mu}g/g$, respectively. These results demonstrate the potential of L. plantarum YS712 as a probiotic culture that can be utilized in the manufacturing of fermentation foods and dietary supplements.

Isolation, Characterization, and Molecular Cloning of the cDNA Encoding a Novel Phytase from Aspergillus niger 113 and High Expression in Pichia pastoris

  • Xiong, Ai Sheng;Yao, Quan-Hong;Peng, Ri-He;Li, Xian;Fan, Hui-Qin;Guo, Mei-Jin;Zhang, Si-Liang
    • BMB Reports
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    • v.37 no.3
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    • pp.282-291
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    • 2004
  • Phytases catalyze the release of phosphate from phytic acid. Phytase-producing microorganisms were selected by culturing the soil extracts on agar plates containing phytic acid. Two hundred colonies that exhibited potential phytase activity were selected for further study. The colony showing the highest phytase activity was identified as Aspergillus niger and designated strain 113. The phytase gene from A. niger 113 (phyI1) was isolated, cloned, and characterized. The nucleotide and deduced amino acid sequence identity between phyI1 and phyA from NRRL3135 were 90% and 98%, respectively. The identity between phyI1 and phyA from SK-57 was 89% and 96%. A synthetic phytase gene, phyI1s, was synthesized by successive PCR and transformed into the yeast expression vector carrying a signal peptide that was designed and synthesized using P. pastoris biased codon. For the phytase expression and secretion, the construct was integrated into the genome of P. pastoris by homologous recombination. Over-expressing strains were selected and fermented. It was discovered that ~4.2 g phytase could be purified from one liter of culture fluid. The activity of the resulting phytase was 9.5 U/mg. Due to the heavy glycosylation, the expressed phytase varied in size (120, 95, 85, and 64 kDa), but could be deglycosylated to a homogeneous 64 kDa species. An enzymatic kinetics analysis showed that the phytase had two pH optima (pH 2.0 and pH 5.0) and an optimum temperature of $60^{\circ}C$.

Characterization of a peroxidase in excretory-secretory product of adult Parasonimus westermani (폐흡충 성충이 분비배설하는 anti-oxidant ensymes의 특성 관찰 및 peroxidase의 정제)

  • Chung, Young-Bae;Kong, Yoon;Cho, Seung-Yull;Kang, Shin-Yong;Choi, Byung-Chan;Lee, Hi-Sung
    • Parasites, Hosts and Diseases
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    • v.31 no.3
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    • pp.259-268
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    • 1993
  • When activity of peroxidase in auld Pnrqfonimn westermqni was monitored using o-dianisidine and $H_2O_2$ as substrates, its specific activity was 1.5 times higher In excretory-secretory product (ESP) than in crude extract. The one was purified by two purification steps of Sephacryl S-300 Superfine gel permeation and DEAE-Trisacryl M anion exchange chromatographies. Its activity increased 16.9 fold with 32.3% recovery. The enzyme was inhibited totally by 1 millimoles of dithiothreitol (DTT), 2-mercaptoethanol and azide. Molecular mass was 16 kDa in reducing SDS-polyacrylamide gel electrophoresis (PAGE) or 19 kDa in TSK-Blue gel filtration high performance liquid chromatography (HPLC). respectively. Special staining for peroxidase by diaminobenzidine on SDS-PAGE confirmed the activity. The peroxidase was less reactive to a paragonimiasis serum when observed by SDS-PAGE/immunoblot. In addition, specific activities of superoxide dismutase (SOD) and catalase were also identified in the ESP. High activities of these antioxidant enzymes in ESP indicate that they are parts of defense mechanisms against reactive oxygen intermediates from host.

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Studies on Some Physiological and Morphological Characteristics of Actinomycetes in Paddy Soil applied Organic Materials (유기물(有機物) 시용시(施用時) 논 토양(土壤)에서 방선균(放線菌)의 형태(形態) 및 생리학적(生理學的) 특성(特性))

  • Park, Kyung-Soo;Park, Youl;Ryu, Jin-Chang
    • Korean Journal of Soil Science and Fertilizer
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    • v.20 no.1
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    • pp.77-84
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    • 1987
  • The population of actinomycetes in paddy soils, with hay compost and inorganic fertilizer had been applied respectively was investigated. Actinomycetes were isolated by using selective medium and the population densities of actinomycetes in paddy soil was examined. The population of actinomycetes were reached at a range from $2.1{\times}10^6$ to $7.4{\times}10^7$ per gram of the soil. The composition of actinomycetes flora changed considerably after hay compost applied. The significant positive correlations between the organic matter content in paddy soil and the actinomycetes populations were given at 1.38 to 2.69 level. According to the result of several morphological observation, similar strains isolated were classified into 21 groups. More detailed taxonomic characterization were carried out on the isolated strains. Therefore, 15 groups of Streptomyces and 6 groups of non-Streptomyces were classified into actinomycetes isolates, percentage of streptomyces and non-Streptomyces strains were 87.2% and 12.8% in the isolated 250 actinomycetes strains respectively. Streptomyces with sporophore of the spiral chain form accounted for 80% of all the Streptomyces isolates. Surface morphology of spores were determined with the electron microscope, three species have a spiny surface, and 13 strains have a smooth spore surface.

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Characterization of Weed Occurrence in Apple Orchards (사과 과수원(果樹園)에서의 잡초발생(雜草發生) 특성(特性)에 관(關)한 연구(硏究))

  • Woo, I.S.;Pyon, J.Y.
    • Korean Journal of Weed Science
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    • v.8 no.2
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    • pp.164-168
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    • 1988
  • In order to get basic information for establishing weed control methods in apple orchards, weed distribution was surveyed at 2 locations in Chungnam Province at 15 days intervals. In apple orchard 48 weed species (18 families) were distributed and Chenopodium album and Polygonum hydropiper were dominated and Digitana sanguinalis, Portulaca oleracea, and Erigeron canadensis were also abundant among them. Highest number of weeds emerged in April and dry weight of weeds were heavy in June and October, especially in summer broadleaf weeds and winter broadleaf weeds, and winter grasses were abundant in spring. Seasonal Shannon's diversity index(H'), maximum diversity(H max'), eveness (J') for the Shannon diversity index, and Simpson index were high in apple orchards. Interspecific competition was more severe than intraspecific competition.

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A Commensal Thermophile, Symbiobacterium toebii: Distribution, Characterization, and Genome Analysis

  • Bae Jin-Woo;Kim Kwang;Song Jae Jun;Ha Jae Seok;Kim Joong-Jae;Kang Gwan-Tae;Kim Mi-Hwa;Hong Seung-Pyo;Sung Moon-Hee
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2001.11a
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    • pp.46-53
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    • 2001
  • A commensal thermophile, Symbiobacterium toebii, isolated from hay compost (toebii) in Korea commensally interacted with a thermophilic Geobacillus toebii sp. nov., which was a new species within the genus Geobacillus on the basis of the phenotypic traits and molecular systematic data. S. toebii required the crude extracts and/or culture supernatant of the Geobacillus toebii for axenic growth and could grow on the temperature between 45 and $70^{\circ}C$ (optimum: $60^{\circ}C$; 2.4 h doubling time) and pH 6.0 and 9.0 (optimum: pH 7.5). The G+C content of the genomic DNA was $65 mol\%$, and the major quinones were MK-6 and MK-7. A phylogenetic analysis of its 16S rDNA sequence indicated that Symbiobacterium toebii was closely related with solely reported Symbiobacterium thermophilum. The presence of the commensal thermophile 16S rDNA and accumulation of indole in all the enriched cultures indicate that Symbiobacterium toebii is widely distributed in the various soils. The genome of S. toebii constituted a circular chromosome of 3,280,275 base pairs and there was not an extra-chromosomal element (ECE). It contained about 4,107 predicted coding sequences. Of these protein coding genes, about $45.6\%$ was encoded well-known proteins and annotated the functional assignment of 1,874 open reading frames (ORFs), and the rest predicted to have unknown functions. The genes encoding thermostable tyrosine phenol-lyase and tryptophan indole-lyase were cloned from the genomic DNA of S. toebii and the enzymatic production of L-tyrosine and L-tryptophan was carried out with two thermostable enzymes overexpressed in recombinant E. coli.

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Identification and Characterization of Paenibacillus polymyxa DY1 Isolated from Korean Soil with New Antibacterial Activity (새로운 항균활성을 보이는 토양 분리 세균 Paenibacillus polymyxa DY1의 분류와 동정)

  • Shin, Eun-Seok;Lee, Hee-Moo;Lee, Bok-Kwon;Kim, Sung-Hoon;Kwon, Sun-Il;Yoo, Kwan-Hee
    • Korean Journal of Microbiology
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    • v.43 no.1
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    • pp.47-53
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    • 2007
  • The DY1 strain of Gram-positive, rod-shaped bacteria was isolated from the soil sample collected from Daeam mountain, Korea. The culture filtrate of DY1 strain showed a broad spectrum of antimicrobial activity on various pathogenic and food poisoning enteric bacterial species tested in vitro. It showed significant growth-inhibitory effect on Salmonella enterica sp., Shigella sp., pathogenic Escherichia coli, Vibrio cholerae, Vibrio parahemolyticus, and Yersinia enterocolitica. For the identification of the DY1 strain, morphological, biochemical and molecular phylogenetic approaches were performed. The DY1 strain was found to be a member of the genus Paenibacillus on the basis of morphological and biochemical analyses. The 16S rDNA of DY1 showed the highest pairwise identity with Paenibacillus polymyxa with 99.79% (1,413 bp/1,416 bp). The antimicrobial entity from DY1 looked different from preciously reported ones and seems to have a great potential to be further studied as a candidate of new antibiotics to control multi-drug resistant pathogens.