• Title/Summary/Keyword: 단백질 동정

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Growth Inhibitory Activity of Enterococcus faecium Isolated from Bovine Intestinal Tract against Enterobacter sakazakii (소 장관 유래 Enterococcus faecium의 Enterobacter sakazakii에 대한 생육저해활성)

  • Park, Ju-Hui;Yoon, Sung-Sik;Park, Young-Seo
    • Food Science of Animal Resources
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    • v.28 no.1
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    • pp.99-104
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    • 2008
  • A lactic acid bacterium showing growth inhibitory activity against Enterobacter sakazakii was isolated from bovine intestinal tracts. From biochemical and molecular biological studies, the isolate was identified and named as Enterococcus faecium JH95. This strain was resistant to kanamycin and streptomycin at a concentration of $100{\mu}g/mL$. E. faecium JH95 had high antimicrobial activity against food-borne pathogens such as Escherichia coli O157:H7, Listeria monocytogenes, Salmonella typhimurium, Staphylococcus aureus, and Clostridium perfrigens. The culture supernatant of this strain did not have antimicrobial activity. The culture broth of this strain failed to show the antimicrobial activity by heat treatment at $100^{\circ}C$ for 5 min or by pretense treatments for 2 hr. This result suggested that the putative antimicrobial substance produced by E. faecium JH95 is likely a protein which is not secreted into culture medium.

Characteristics of Pichia anomala K15 Producing Killer Toxin Isolated from Traditional Nuruk (전통 누룩으로부터 분리된 Killer Toxin 생산 균주 Pichia anomala K15의 특성)

  • Jung, Hee-Kyoung;Park, Chi-Duck;Lee, Gee-Dong;Park, Seung-Chun;Park, Hwan-Hee;Hong, Joo-Heon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.8
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    • pp.1077-1082
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    • 2007
  • In this study, killer yeasts were isolated from traditional Nuruk to improve storage and suppress contaminant in food industry. Among killer yeasts, yeast K15 showed strong killer toxin activity and inhibited growth of Salmonella Typhimurium and Vibrio parahaemolyticus. Killer yeast K15 was identified with Pichia anomala by the Microlog TM 4.0 identification system and homology of the ITS sequence. Killer toxin generated from P. anomala K15 was inactivated by pronase E and suggested to be a protein. Therefore killer toxin of P. anomala K15 was thought to be safe in human such as bacteriocin. P. anomala K15 was sufficient for growth in 50% glucose and could be used to prevent contaminant in initial stages of alcohol beverage fermentation.

Isolation and Identification of Fibrinolytic Bacteria from Korean Traditional Chungkookjang (전통식품(청국장)으로 부터 fibrin용해 세균의 분리 동정)

  • Heo, Seok;Joo, Hyun-Kyu;Lee, Si-Kyung
    • Applied Biological Chemistry
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    • v.41 no.2
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    • pp.119-124
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    • 1998
  • In this study, the bacteria which could hydrolyze the fibrin produced through the blood coagulation mechanism in the human body, were isolated from Chungkookjang. The KCK-7 strain was selected among the isolated bacteria as the best strain for fibrinolytic activity. It was spore forming and Gram positive. $C_{150}$ anteiso fatty acid and $C_{150}$ iso fatty acid were 40.85% and 19.47%, respectively as major component among its cellular fatty acid composition. It showed the similarity of 63.6%, compared with standard strain. It was thus identified to be Bacillus subtilis according to Bergey's manual of systematic bacteriology and its fatty acid profiles af Gas chromatography. The optimum culture temperature and pH were $37^{\circ}C$ and 8 for the production of fibrinolytic enzyme by Bacillus subtilis KCK-7.

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The Changes of Quantity and Quality of Proteins in Medium and Cytoplasm during In Vitro Maturation of Bovine Oocytes (한우 난포란의 체외성숙 배지와 세포질 내의 단백질 변화)

  • Park Y. S.;Park H. Y.
    • Reproductive and Developmental Biology
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    • v.29 no.3
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    • pp.187-191
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    • 2005
  • This study was designed to investigate the changes of quantity and quality of proteins in medium and cytoplasm during in vitro maturation of bovine oocytes. The total quantity of proteins in medium decreased from 0 to 4.5 hr, but increased from 13.5 to 18 hr after the onset of in vitro maturation. The total quantity of protein in cytoplasm increased from 0 to 4.5 hr, decreased from 4.5 to 9 hr, and increased after 18 hr after the onset of in vitro maturation. A total of 298 protein spots was detected on a gel of 2D SDS-PAGE form maturation medium. Among 28 protein spots expressed significant differences in their quantity, 8 proteins were identified by peptide mass fingerprinting (aldose reductase, alpha enolase, apolipoprotein A-1 precursor, 43kDa collectin precursor, heat shock 27kDa protein, plasminogen activator inhibitor-1 precursor, thrombospondin 1, transitional endoplasmic reticulum ATPase). Among total of 35 protein spots detected on gel of 2D SDS-PACE from oorytes cytoplasm, $\beta$-tubulin was identified by peptide mass fingerprinting.

Viral Hemorrhagic Septicemia Virus NV Gene Decreases Glycolytic Enzyme Gene Transcription (바이러스성 출혈성 패혈증 바이러스 NV 단백질에 의한 glucokinase 전사 활성의 억제)

  • Cho, Mi Young;Hwang, Jee Youn;Ji, Bo Young;Park, Myoung Ae;Seong, Mi So;Kim, So Young;Jung, Ye Eun;Cheong, Jae Hun;Choi, Yung Hyun
    • Journal of Life Science
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    • v.26 no.12
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    • pp.1470-1476
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    • 2016
  • The viral hemorrhagic septicemia virus (VHSV), which belongs to the Novirhabdovirus genus of the Rhabdoviridae family, is a viral pathogen that causes severe losses in the olive flounder farming industry. Among six encoding VHSV proteins, the non-virion (NV) protein has been shown to have an impact on virulence. In our previous studies, transcriptomics microarray analysis by using VHSV-infected olive flounder showed that VHSV infection significantly down-regulated the mRNA expression of glycolytic enzymes. In addition, VHSV NV protein variants decreased the intracellular ATP level. Based on these results, we have tried to examine the effect of VHSV NV protein on glycolytic enzyme glucokinase expression, which phosphorylates glucose to glucose 6-phosphate. Our results indicated that the NV protein significantly decreased the mRNA expression of glucokinase in olive flounder HINAE cells. Furthermore, the NV protein played a negative role in the promoter activation of glucokinase. Furthermore, glucose uptake was effectively inhibited by VHSV infection and NV protein expression in olive flounder HINAE cells. These results suggest that the VHSV NV protein negatively regulates glycolytic enzyme expression by a transcription level and eventually leads to gradual morbidity of olive flounder through cellular energy deprivation. The present results may be useful for the prevention and diagnosis of VHSV infection in olive flounder.

Identification of Rhizobium fredii and Bradyrhizobium japonicum by Polyacrylamide Gel Electrophoresis (전기영동법(電氣泳動法)에 의(依)한 대두(大豆) 근류균(根瘤菌) Rhizobium fredii와 Bradyrhizobium japonicum의 분류(分類) 및 동정(同定))

  • Yun, Han-Dae;Cho, Moo-Je;Lee, Ke-Ho
    • Applied Biological Chemistry
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    • v.30 no.2
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    • pp.163-168
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    • 1987
  • A method, based upon the separation of cellular proteins by one-and two-dimensional electrophoresis was used for distinguishing butween Bradyrhizobium japonicum strains and Rhizobium fredii strains. Significant differences in protein pattern of one-dimensional SDS-PAGE vs-ere observed between Rhizobium fredii strains and Bradyrhizobium japonicum strains. The differences in six distinct main lands were observed among total 52 kinds of protein bands. Furthermore, the distribution of proteins in two groups by two-dimensional polyacrylamide gel electrophoresis was very different. The majority of visible proteins of Rhizobium fredii were acidic, whereas those of Bradyrhizobium japonicum were basic. In addition, amino acid composition was analyzed to detect the differences between two groups. No significant differences in amino acid composition were observed between Bradyrhizobium japonicum strains and Rhizobium fredii strains. The results indicate that one-and two-dimensional polyacrylamide gel electrophoresis were useful for identifying rhizobia isolates. One-dimensional SDS-PAGE of rhizobia proteins provided a rapid method for screening a large number of isolates, whereas two-dimensional electrophoresis was more of resolution and easiness for analyzing protein spots.

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Separation Characteristics of IgY (Immunoglobulin Yolk) in Various HPLC Columns (다양한 HPLC Column에서의 IgY(Immunoglobulin Yolk) 분리특성)

  • Song, Sung Moon;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.50 no.4
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    • pp.659-665
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    • 2012
  • IgY (Immunoglobulin Yolk) in egg yolk corresponds to IgG (Immunoglobulin G) in animal serum and plays an important role as immunological proteins in intestines. Carrageenan and Arabic gum were used as pretreatment agents to purify IgY from fresh egg yolk. DEAE (Diethylaminoethyl) Sepharose column in FPLC (Fast Protein Liquid chromatography) was an ion exchange tool to remove contaminants as well as to elute IgY from the column. GF HPLC (Gel Filtration High Performance Liquid Chromatography) enables to measure the molecular weights of IgY and to identify the purified IgY by comparing the molecular weight of standard IgY with the purified one. IgY is a heterogeneous group of different molecular weight and ionic properties, which was investigated with various IE HPLC (Ion Exchange High Performance Liquid Chromatography) columns such as AX, CX and SCX. Three peaks of IgY were separated in the AX column under the conditions of 0.5 M NaCl and pH=8. The SCX column also gave the three peaks of IgY at 0.5 M NaCl and pH=5.

Characterization of Extracellular Proteolytic Enzyme of Isolated Psychrotrophic Bacteria from Cheddar Cheese (체다치즈에서 분리한 내냉성미생물의 단백질분해효소의 특성)

  • Kim, Eun-Ah;Lee, Kyung-Wook;Boo, Won-Back;Lee, Hyung-Hoan;Kwak, Hae-Soo
    • Korean Journal of Food Science and Technology
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    • v.23 no.4
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    • pp.452-458
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    • 1991
  • Psychrotrophs producing protease were isolated during ripening periods of Cheddar cheese and one of them containing the highest protease activity was identified as Pseudomonas fluorescens 65. The extracelluar proteolytic enzyme was partially purified from P. fluorescens 65 through the Sephadex G-100 gel filtration. The protease was eluted between 190 ml and 230 ml of elution volume of sodium phosphate buffer. The purified protease showed a single band in SDS-PAGE and its molecular weight was 47,000. The composition of amino acid for the protease was determined and the most abundant amino acids were glutamic acid (14.96%) and serine (13.86%). The optimum temperature and pH for the activity was $45{\sim}50^{\circ}C$ and 6.0, respectively.

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First Report of Cucumber mosaic virus Isolated from Sambungai (Gynura procumbens) (명월초에서 분리한 오이모자이크바이러스의 감염 첫 보고)

  • Kwon, June;Hong, Jin-Sung
    • Research in Plant Disease
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    • v.23 no.4
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    • pp.379-382
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    • 2017
  • In March 2016, an isolate of Cucumber mosaic virus (named Gyp-CMV) was isolated from the Sambungai (Gynura procumbens) showing the symptoms of mosaic and chlorosis. The isolate Gyp-CMV was characterized by disease reactions in several indicator plants, reverse transcription-polymerase chain reaction (RT-PCR), PCR-restriction fragment length polymorphism, and sequence analysis of movement protein (3a) and coat protein (CP) genes. Tobacco, tomato, pepper, ground cherry, and lambsquarters (Chenopodium quinoa and C. amaranticolor) appeared typical CMV symptoms, but zucchini and cucumber were not infected. Phylogenetic analysis of the 3a and CP gene indicated that Gyp-CMV belongs to the CMV subgroup II. Sequence identities of the Gyp-CMV 3a and CP genes showed 99.3% and 100% to that of Hnt-CMV at amino acid level. To our knowledge, this is the first report of CMV infection in Gynura procumbens.

Isolation and Selection of Functional Microbes for Eco-friendly Turfgrass Management in Golf Course from Livestock Manure Compost (친환경 잔디관리를 위한 가축분퇴비 중 기능성미생물의 분리 및 선발)

  • Jeong, Je-Yong;Kim, Young-Sun;Cho, Sung-Hyun;Lee, Geung-Joo
    • Weed & Turfgrass Science
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    • v.6 no.2
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    • pp.157-164
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    • 2017
  • Functional microorganisms decompose various organic matter by enzyme activity and suppress plant disease caused by pathogen. This study was conducted to isolate and select functional microorganisms with protein or carbohydrate degradation activities and antagonistic activity against turfgrass fungal pathogens for eco-friendly turfgrass management in golf course from compost containing livestock manure of poultry or swine. Totally 68 isolates collected from livestock manure compost strains were isolated and tested for their activities of amylase, protease and lipase and antagonistic activities against Rhizoctonia solani AG2-2, R. solani AG1-1, and Sclerotinia homoeocarpa. Among the isolates, 34 strains were selected as functional microbes showing higher activities of amylase and protease. Three isolates of ASC-14, ASC-18, and ASC-35 among the 34 strains were selected as antifungal bacterial strains repressing the above 3 turfgrass fungal pathogens. Analysis results of 16s rRNA gene sequence and phylogenic cluster indicated that ASC-14 and ASC-18 belonged to Bacillus amyloliquefaciens, while ASC-35 was B. subtilis, respectively.