• Title/Summary/Keyword: 단백질 동정

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Isolation and identification of protease-producing bacteria from the intertidal zone in Jeju Island, Korea (제주 조간대로부터 단백질 가수분해효소를 생산하는 세균의 분리 및 동정)

  • Moon, Young-Gun;Dharaneedharan, Subramanian;Kim, Dong-Hwi;Park, So-Hyun;Heo, Moon Soo
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.382-388
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    • 2015
  • Eleven protease-producing bacteria were isolated from the organisms' external shells and the inorganic materials collected from intertidal zone of Jeju Island, Republic of Korea. The samples were diluted serially, inoculated on Zobell agar plates with 1% skim milk and incubated at $20^{\circ}C$. Clear zone forming colonies were selected as protease-producing bacteria and each strain was identified based on the phylogenetic analysis with their 16S rDNA sequences. Strains JJM125, JJM129, YG47 and YG49 belong to the marine bacterial genus Pseudoalteromonas; strain JJM122 belong to the genus Microbulbifer; strains YG51, YG52, YG62 and YG63 belong to the genus Vibrio; and strain YG65 belong to genus Bacillus. Hence, the present study suggests that these protease producing bacteria could be further used to develop new varieties of protease with various biotechnological applications.

Studies on the Changes of Protein, Peptide and Amino Acid During Natto Preparation (납두제조중(納豆製造中)의 단백질(蛋白質), Peptide 및 AminoAcid의 변화(變化)에 관(關)한 연구(硏究))

  • Kim, Su-Yung;Kim, Ze-Uook
    • Applied Biological Chemistry
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    • v.8
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    • pp.11-20
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    • 1967
  • In order to study the change of nitrogeneous compounds during the "Natto" preparation, the contents of insoluble protein, water soluble protein and amino nitrogen were determined and the pattern of peptides and amino acids was investigated by paper chromatography for the fractions resulting from molecular sieving. The results are summarized as follows: 1. Insoluble protein nitrogen which was increased to 84% by autoclaving the native soybean decreased to 44%, whereas the trichloroacetic acid soluble nitrogen increased from 8% to 45% during Natto preparation, But the soluble protein nitrogen showed a slight increase. 2. Fractionation of the peptides using Dowex-50 resins showed that they consisted mostly of lower molecular weight peptides which increased in accordance with the progress of fermentation, especially after 30-hour period. 3. Sixteen known and two unknown amino acids, and three peptides with different Rf values were identified during the Natto preparation. Their appearance showed some difference in that phenylalanine appeared after 10 hours, methionine, after 20 hours and proline, after 30 hours, respectively. The three peptides appeared at the different stage of fermentation.

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Effects of Some Plant Growth Regulators on Protein Biosynthesis of Carrot Cells (당근 세포의 단백질 생합성에 대한 몇가지 식물생장조절제의 영향)

  • Yoo, Ki-Jung;Park, Chang-Kyu;Song, Tae-Chul
    • Applied Biological Chemistry
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    • v.28 no.3
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    • pp.187-195
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    • 1985
  • Electrophoretic studies of protein extracts from carrot calluses suspension-cultured on the media containing kinetin, BA, IAA, NAA or $GA_3$ at the levels of $10^{-6},\;10^{-5},\;10^{-4}M$, respectively, were performed to identify polypeptides and proteins regulated by auxin, cytokinin or GA. Fifteen bands of polypeptide(s) were observed in the callus cultured in the control medium devoid of growth regulators, and their molecular weights were $18._4,\;20._2,\;20._0,\;34._9,\;35._7,\;37._4,\;40._3,\;42._2,\;44._1,\;44._4,\;49._3,\;55._0,\;56._6,\;58._1,\;and\;59._9\;KD$, respectively. The synthesis of polypeptide appeared to be promoted in two bands by kinetin, in six bands by BA, in one band by IAA, in two bands by NAA, and in four bands by $GA_3$, while inhibited in five bands by kinetin, in three bands by BA, in four bands by IAA, in three bands by NAA and in three bands by $GA_3$. The polypeptides of $40._3\;KD\;42._2\;KD$ seemed to be regulated by cytokinins, and those of $44._1\;KD,37._4\;KD,\;and\;56._6\;KD$ by auxins. The proteins of three bands with relative mobilities of 0.56, 0.84, and 0.92, respectively, increased in the calluses cultured on the media containing kinetin, IAA, $GA_3$, NAA or BA, compared to the control, but it was difficult to identify the proteins specific for each growth regulator.

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Proteomic Responses of Diploid and Tetraploid Roots in Platycodon grandiflorum (2배체와 4배체 도라지의 단백질 발현양상 비교 분석)

  • Kim, Hye-Rim;Kwon, Soo-Jeong;Roy, Swapan Kumar;Cho, Seong-Woo;Kim, Hag-Hyun;Moon, Young-Ja;Boo, Hee-Ock;Woo, Sun-Hee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.3
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    • pp.394-400
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    • 2015
  • The roots of Platycodon grandiflorum species either dried or fresh, are used as an ingredient in salads and traditional cuisine in Korea. To interpret the root proteins, a systematical and targeting analysis were carried out from diploid and tetraploid roots. Two dimensional gels stained with CBB, a total of 39 differential expressed proteins were identified from the diploid root under in vivo condition using image analysis by Progenesis Same Spot software. Out of total differential expressed spots, 39 differential expressed protein spots (${\geq}\;1.5$-fold) were analyzed using LTQ-FTICR mass spectrometry. Except two proteins, the rest of the identified proteins were confirmed as down-regulated such as Isocitrate dehydrogenase, Proteasome subunit alpha type-2-B. However, the most of the identified proteins from the explants were mainly associated with the oxidoreductase activity, nucleic acid binding, transferase activity and catalytic activity. The exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.

Inhibitory Effects of a Recombinant Viral Cystatin Protein on Insect Immune and Development (바이러스 유래 시스타틴 재조합 단백질의 곤충 면역 및 발육 억제효과)

  • Kim, Yeongtae;Eom, Seonghyun;Park, Jiyeong;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.53 no.4
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    • pp.331-338
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    • 2014
  • Cystatins (CSTs) are reversible and competitive inhibitors of C1A cysteine proteases, corresponding to papain-like cathepsins in plants and animals. A viral CST (CpBV-CST1) was identified from a polydnavirus, Cotesia plutellae bracovirus (CpBV). Our previous study indicated that a transient expression of CpBV-CST1 interfered with immune response and development of Plutella xylostella larvae. To directly demonstrate the protein function, this study produced a recombinant CpBV-CST1 protein (rCpBV-CST1) using bacterial expression system to determine its inhibitory activity against cysteine protease and to assess its physiological alteration in insect immune and development. The open reading frame of CpBV-CST1 encodes a polypeptide of 138 amino acids (${\approx}15kDa$). rCpBV-cystatin protein in BL21 STAR (DE3) competent cells containing a recombinant pGEX4T-3:CpBV-CST1 was over-expressed by 0.5 mM IPTG for 4 h. In biological activity assay, the purified rCpBV-CST1 showed a significant inhibition against papain activity. It inhibited a cellular immune response of hemocyte nodule formation in the beet armyworm, Spodoptera exigua. Moreover, its oral administration retarded larval development of the diamondback moth, Plutella xylostella in a dose-dependent manner. These results suggest that CpBV-CST1 may be applied to control insect pest populations.

A Potent Tissue Destructive Activity of Secreted Proteins of Aeromonas hydrophila (조직 괴사 활성을 지닌 Aeromonas hydrophila 의 분비 단백질에 관한 연구)

  • Kim, Kyu Lee;Choe, Yunjeong;Kang, Ho Young
    • Journal of Life Science
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    • v.25 no.2
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    • pp.214-222
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    • 2015
  • Aeromonas hydrophila is the most common water fish pathogen and cause diseases such as hemorrhagic septicemia, dropsy, ulceration and asymptomatic septicemia. A. hydrophila secretes many extracellular products (ECPs) which contribute to effective infection, wide distribution and great adaptability to environmental changes. Crude ECPs of A. hydrophila CK257, a strain used in this study, exhibits a toxic activity to the animals including mouse, rabbit and fish. Toxic symptoms were indicated by tissue damage and skin injuries in animal. When ECPs were subcutaneously injected to animals, skin damages were observed, appearing like necrosis. Preliminary research demonstrated that the active factors are protein component. The crude ECPs were collected after ammonium sulfate precipitation of cell-free culture supernatant. ECPs were fractionated with the use gel filtration chromatography. Five ECP fractions were obtained, of which one fraction was found to be toxic to goldfish. MALDI-TOF analyses provided two interesting proteases called M35 and M28. Both M35 and M28 are known as metalloprotease. Accordingly, proteins in an active fraction exhibited caseinolytic activity. These proteins were difference of caseinolytic activity under different metallic ions. Also active fraction has elastolytic activity. These results suggested that peptidase M28 and M35 may be a candidate factor for tissue necrosis activity about infection with A. hydrophila.

Changes of the Protein Contents of Seafood Cooking Drips by Gamma Irradiation (감마선 조사에 의한 수산 자숙액의 단백질 함량 변화)

  • Choi, Jong-Il;Kim, Hyun-Joo;Sung, Nak-Yun;Byun, Eui-Baek;Kim, Jae-Hun;Chun, Byung-Soo;Ahn, Dong-Hyun;Cho, Kook-Yeon;Byun, Myung-Woo;Lee, Ju-Woon
    • KSBB Journal
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    • v.23 no.6
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    • pp.489-493
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    • 2008
  • Although the seafood cooking drips were the byproducts from the fishery industry and being wasted, it had many nutrients including proteins. In this study, the effect of a gamma irradiation on the cooking drips from Hizikia fusiformis, Enteroctopus dofleni and Thunnus thynnus were investigated. The cooking drips were extracted with 70% ethanol solution, and the extracts were analysed for the protein concentration by three different methods of Lowry, BCA and Kjeldahl. The extracts were irradiated with different doses and the protein contents were compared with respect to the absorbed doses. Total content of the proteins was increased with increasing irradiation dose. The change of protein pattern in the irradiated cooking drips was also confirmed by SDS-PAGE analysis. These results shown that the proteins in cooking drips could be unfolded or aggregated by the irradiation. Therefore, gamma irradiation could be considered as an effective method for extracting useful proteins.

Characterization of Antimicrobial Substance Produced by Lactobacillus paraplantarum KNUC25 Isolated from Kimchi (김치로부터 분리된 Lactobacillus paraplantarum KNUC25가 만드는 항균 물질의 특성)

  • Kim, Ma-Rie;Lee, Su-Jin;Seul, Keyung-Jo;Park, Yu-Mi;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.37 no.1
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    • pp.24-32
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    • 2009
  • The KNUC25 strain isolated from over-fermented whole Chinese cabbage kimchi was examined for its physiological characteristics using API 50 CHL system assay and identified as Lactobacillus paraplantarum by analysis of whole-cell protein SDS-PAGE pattern assay and similarity of 16S rDNA sequence. L. paraplantarum KNUC25 had a broad antimicrobial activity spectrum from Gram positive to Gram negative bacteria. Scanning electron micrograph analysis showed that KNUC25 might attack to cell surface of indicator cells and destruction can lead to inhibition of the cell growth. The antimicrobial substance of the KNUC25 strain was stable to various degrading enzymes and at high temperature and not a plasmid-born matter. Resistance to proteolytic enzymes showed that an antimicrobial activity of KNUC25 might not be caused by proteinous substance. Maximum production of antimicrobial substance was the exponential growth phase at $30^{\circ}C$.

Comparative Analysis of Genetic Variation of Cucumber Mosaic Virus from Commelina communis in Korea (국내 닭의장풀에서 분리된 오이모자이크바이러스 분리주들의 외피단백질 유전자와 병징 다양성 비교)

  • Park, Tae-Seon;Hong, Jin-Sung
    • Research in Plant Disease
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    • v.24 no.2
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    • pp.170-173
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    • 2018
  • Three isolates of Cucumber mosaic virus isolated from Commelina communis plants showing chlorosis and mosaic were collected in Chungju and Chuncheon, Korea. To confirm genetic variation of these three isolates (CMV-Co, CMV-Co2, and CMV-Co3), we performed PCR-RFLP and sequence analysis. Sequences of coat protein genes of CMV-C0, -Co2 and -Co3 were compared with CMV-Fny and showed 96.3%, 96.3%, and 95.9% similarities, respectively. In host reactions, three CMV-Co isolates induced systemic necrosis in Cucurbita pepo unlike CMV-Fny and CMV-Co, CMV-Co2 and CMV-Co3 observed differential symptoms responses in Physalis angulata and Nicotiana rustica. These results indicated that three isolates of CMV isolated from C. communis have genetic and biological variation.

Purification and Characterization of Xylanase from Fomitopsis palustris in Rice Straw Culture (볏짚분해과정 중에 생산하는 Fomitopsis palustris 균체 외 Xylanase의 분리정제 및 효소특성)

  • Yoon, Jeong-Jun;Lee, Young-Min;Choi, Doo-Yeol;Kim, Young-Kyoon;Kim, Yeong-Suk
    • Journal of the Korean Wood Science and Technology
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    • v.35 no.6
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    • pp.159-165
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    • 2007
  • An extracellular xylanase from the brown-rot fungus Fomitopsis palustris grown on rice straw culture was purified to a single protein band. On SDS-PAGE, the molecular mass of purified enzyme was estimated to be about 43 kDa. The amino acid sequence of the proteolytic fragments showed high homology with fungal glycoside hydrolase family 10 xylanases. The $K_m$, $K_{cat}$ and $V_{max}$ for birch xylan were $31mg/m{\ell}$, $2.3{\times}10^4/min$ and 252.3 U/mg, respectively. The optimal activity of the purified xylanase from F palustris was observed at pH 4.0~5.0 and $70^{\circ}C$.