• Title/Summary/Keyword: Protein fractions

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Characteristics and purification of proteoglycan from Phellinus igniarius (Phellinus igniarius로부터 분리한 단백다당류의 분리 및 특성)

  • Kim, Seon-Hee;Jung, In-Chang;Kwon, Yong-Il;Kim, So-Yeun;Lee, Jong-Suk;Lee, Hang-Woo;Lee, Jae-Sung
    • Applied Biological Chemistry
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    • v.43 no.1
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    • pp.57-62
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    • 2000
  • The proteoglycan, intracellular and extracellular, extracted from the liquid culture of Phellinus igniarius were purified and characterized. The mycelial productivity was proved to be better in shaking culture compared to standing culture. The productivity of intracellular proteoglycan of Phellinus igniarius appeared to be similar in two culturing methods. The standing culture of Phellinus igniarius produced 6 times as much extracellular proteoglycan compared to shaking culture. The proteoglycan were purified to a single peak by ion exchange chromatography(DEAE-cellulose) followed by gel filtration(Sepharose 2B). PIEPDG contained 79.0% total sugar and 7.2% protein. PIEPAG contained 56.7% total sugar and 40.8% protein. PIIPDG contained 64.8% total sugar and 17.4% protein. PIIPAG contained 56.9% total sugar and 41.5%n protein. The molecular weights of all the fractions were estimated to be above 100,000, from 134KDa of PIEPDG to 560 KDa of PIEPAG. The results of sugar analysis by HPLC showed that PIEPDG contains glucose only. The sugar part of PIIPDG and PIIPAG were consisted of glucose and inositol. The PIEPAG contained three kinds of monosaccharides, glucose, fructose and inositol.

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Cellular responses and proteomic analysis of hemolytic Bacillus cereus MH-2 exposed to epigallocatechin gallate (EGCG) (Epigallocatechin Gallate (EGCG)에 노출된 용혈성 Bacillus cereus MH-2의 세포 반응 및 프로테옴 분석)

  • Kim, Dong-Min;Park, Sang-Kook;Oh, Kye-Heon
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.260-268
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    • 2016
  • The aim of this work was to investigate the cellular responses and proteomic analysis of Bacillus cereus MH-2 exposed to EGCG. Strain MH-2 was isolated from commercial Ssamjang and has the hemolytic activity. Survival of the MH-2 strain with time in the presence of different concentrations of EGCG under sublethal conditions was monitored. The amount of alginate from MH-2 strain decreased depending on the increasing concentrations of EGCG and increased depending on the exposure time at any particular EGCG concentration. Analysis of SDS-PAGE and Western blot using anti-DnaK and anti-GroEL revealed that two stress shock proteins, 70 kDa DnaK and 60 kDa GroEL were found to decrease in proportion to the EGCG concentration in exponentially growing cultures. Scanning electron microscopic analysis demonstrated the presence of protrusions and fused rod forms on the cells treated with EGCG. 2-DE of soluble protein fractions from MH-2 cultures showed 20 protein spots changed by EGCG exposure. These proteins involved in enterotoxins (hemolysin BL lytic component L1 and hemolysin BL-binding protein), chaperons (DnaK and GroEL), cell defense (peptidase M4 family proteins), and various biosynthesis and energy metabolism were identified by peptide mass fingerprinting using MALDI-TOF. These results provide clues for understanding the mechanism of EGCG-induced stress and cytotoxicity on B. cereus MH-2.

Isolation and Partial Physicochemical Characterization of Bile Acid-Binding Fraction from Rice Bran Protein Hydrolysates (미강 단백질 가수분해물에서 담즙산 결합 획분의 분리 및 특성구명)

  • Cho, Wan-Il;Moon, Tae-Wha
    • Korean Journal of Food Science and Technology
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    • v.29 no.3
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    • pp.417-426
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    • 1997
  • Rice bran protein hydrolysates were prepared and some of their physicochemical properties were investigated to utilize rice bran as starting material for functional food ingredient. Rice bran proteins (RBP) were prepared from defatted rice bran by alkaline extraction and isoelectric precipitation. The enzyme for hydrolysis of RBP was selected through measuring relative activity by pH-drop method and comparing the degree of hydrolysis (DH) of hydrolysates. The enzymatic hydrolysates prepared by $Esperase^{\circledR}$ treatment were partitioned into two fractions by ultrafiltration(UF) with a 10 kDa molecular weight cut-off membrane. Each fraction was applied to a cholic acid-conjugated ${\omega}-aminohexyl$ Sepharose 4B column and the bile acid-binding components were obtained by eluting with deoxycholate. Gel permeation chromatography on a Sephadex G-50 column revealed that molecular weight of the bile acid-binding fraction of UF permeate was distributed in ranges of $2\;kDa{\sim}10\;kDa$ and $0.2\;kDa{\sim}0.6\;kDa$. Three peaks (R-1, R-2 and R-3) were obtained by prep-HPLC of bile acid-binding fraction of UF retentate and analyzed for total and free amino acid composition. The results showed that proline content of the bile-acid binding polypeptides and peptides was four times as much as that of rice bran protein and that the peak corresponding to higher average hydrophobicity had a higher free amino acid content. Average hydrophobicity slightly increased with enzymatic hydrolysis.

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The Effect of Aster glehni Fr. Schm. Extracts on Whitening and Anti-Wrinkle (섬쑥부쟁이(Aster glehni Fr. Schm.) 분획물의 미백 및 주름 개선 효과)

  • Kim, Han-Hyuk;Park, Gun-Hye;Park, Kang-Soo;Lee, Jin-Young;Kim, Tae-Hoon;An, Bong-Jeun
    • Journal of Life Science
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    • v.20 no.7
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    • pp.1034-1040
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    • 2010
  • Plants and their extracts can be utilized as inexpensive and rich resources of active constituents in the cosmetic field, as well as the food, pharmaceutical and medicinal fields. Until now, Aster glehni Fr. Schm. had no known active effect, except on anti-oxidation, that was found during investigations for application in the cosmetic field. In this study, we examined the inhibition of enzymatic reactions to protein levels in inclusive B16F10 melanoma cell lines. Significant inhibition of enzymatic reactions was observed in the EtOAc extract, which advanced to tyrosinase protein and TRP-1 in the B16F10 melanoma cell line. These results indicated that the effect of EtOAc extract inhibited expressions of tyrosinase protein and TRP-1 in the B16F10 melanoma cell line by 30.5% and 41.5% at 100ug/ml respectively. On the other hand, antimicrobial activity was evaluated to the four fractions in normal flora of the skin. Hexane extract was only exhibited in the higher clear zone in all strains. In conclusion, any cosmeceutical effects of Aster glehni Fr. Schm. may have a potential meaning, as well as possible value for further studies regarding the effects of chemical constituents of Aster glehni Fr. Schm.

Enhanced functional and structural properties of high-density lipoproteins from runners and wrestlers compared to throwers and lifters

  • Lee, Hwa-Hyung;Park, Jeong-Euy;Choi, In-Ho;Cho, Kyung-Hyun
    • BMB Reports
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    • v.42 no.9
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    • pp.605-610
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    • 2009
  • Plasma high-density lipoprotein cholesterol (HDL-C) levels are inversely correlated with the risk of cardiovascular disease, and are known to increase with repetitive exercise. In the current study, HDL fractions from athletes' sera were isolated and compared as a function of the type of sport (runners [n = 10], throwers [n = 10], wrestlers [n = 10], and weight lifters [n = 8]), and as an age- and gender-matched reference group (n = 14). Among athletes, HDL from runners had the strongest antioxidant activity. Immunodetection showed that runners and wrestlers had the highest levels of apoA-I and lowest levels of apoA-II in their HDL. Electron microscopy also revealed that HDL2 of runners and wrestlers were the largest in size. In conclusion, although all athlete groups had significantly better serum lipid/lipoprotein profiles than the reference group, runners and wrestlers had the most desirable lipoprotein function and structure, including antioxidant activity, HDL-associated enzyme activities and increased particle size.

Transposable Elements and Genome Size Variations in Plants

  • Lee, Sung-Il;Kim, Nam-Soo
    • Genomics & Informatics
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    • v.12 no.3
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    • pp.87-97
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    • 2014
  • Although the number of protein-coding genes is not highly variable between plant taxa, the DNA content in their genomes is highly variable, by as much as 2,056-fold from a 1C amount of 0.0648 pg to 132.5 pg. The mean 1C-value in plants is 2.4 pg, and genome size expansion/contraction is lineage-specific in plant taxonomy. Transposable element fractions in plant genomes are also variable, as low as ~3% in small genomes and as high as ~85% in large genomes, indicating that genome size is a linear function of transposable element content. Of the 2 classes of transposable elements, the dynamics of class 1 long terminal repeat (LTR) retrotransposons is a major contributor to the 1C value differences among plants. The activity of LTR retrotransposons is under the control of epigenetic suppressing mechanisms. Also, genome-purging mechanisms have been adopted to counter-balance the genome size amplification. With a wealth of information on whole-genome sequences in plant genomes, it was revealed that several genome-purging mechanisms have been employed, depending on plant taxa. Two genera, Lilium and Fritillaria, are known to have large genomes in angiosperms. There were twice times of concerted genome size evolutions in the family Liliaceae during the divergence of the current genera in Liliaceae. In addition to the LTR retrotransposons, non-LTR retrotransposons and satellite DNAs contributed to the huge genomes in the two genera by possible failure of genome counter-balancing mechanisms.

Molecular Characterization and Bitter Taste Formation of Tryptic Hydrolysis of 11S Glycinin

  • Kim, Mi-Ryung;Choi, Sang-Yun;Lee, Cherl-Ho
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.509-513
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    • 1999
  • The molecular size reduction and the formation of bitterness during a tryptic hydrolysis of soybean 11S glycinin were determined by using quantitative analysis and organoleptic evaluation. The 11S glycinin of 90% purity was prepared by cryoprecipitation and Con A Sepharose 4B affinity chromatography, and hydrolyzed with trypsin in a pH-stat reactor for 4 h. Bitterness was formed within 1 h of hydrolysis, and then slowly increased up to $3.5\times10^{-5}$ M quinine-HCl equivalent. The extent of hydrolysis (DH) was 7% at 1 h and increased up to 12% by the end of the reaction. The -amino nitrogen content increased from an initial 0.7 mM to 7 mM at the end of the period. The SDS-PAGE analysis showed that the acidic subunit of 11S glycinin was mostly hydrolyzed. The GP-HPLC analysis indicated that the bitterness was mainly contributed by the peptide fractions of molecular weights of 360-2,100 Da.

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Effect of Nordihydroguaiaretic Acid on the Secretion of Lipoprotein Lipase

  • Kim, Sun-Mee;Park, Tae-Won;Park, Jin-Woo
    • BMB Reports
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    • v.35 no.5
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    • pp.518-523
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    • 2002
  • Nordihydroguaiaretic acid (NDGA), an inhibitor of lipoxygenase, inhibits the secretion of proteins and causes the redistribution of resident Golgi proteins into the endoplasmic reticulum (ER). In this study, the effect of NDGA on lipoprotein lipase (LPL) secretion was investigated in 3T3-L1 adipocytes, and compared with those of brefeldin A (BFA), a well-known fungal metabolite that exhibits similar ER-Golgi redistribution. Both BFA and NDGA blocked secretions of LPL. In the presence of BFA, the active and dimeric LPL was accumulated in adipocytes. After endoglycosidase H (endo H) digestion, the proportion of LPL subunits with partially endo H-sensitive oligosaccharide was significantly increased with BFA. However, in the presence of NDGA, the cellular LPL became inactive, and only the endo H-sensitive fraction of the LPL subunit was observed. An increase of the aggregated forms was observed in the fractions of the sucrose-density gradient ultracentrifugation. These properties of LPL in the NDGA-treated cells were similar to those of LPL that is retained in ER, and the effects of NDGA could not be reversed by BFA. These results indicate that the inhibitory mechanism of NDGA on the LPL secretion is functionally different from the ER-Golgi redistribution that is induced by BFA.

Biochemical Changes in Sorghum Leaves Infected with Leaf Spot Pathogen, Drechslera sorghicola

  • Khan, A.J.;Deadman, M.L.;Al-Maqbali, Y.M.;Al-Sabahi, J.;Srikandakumar, A.;Rizvi, S.G.
    • The Plant Pathology Journal
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    • v.17 no.6
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    • pp.342-346
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    • 2001
  • The physiological changes in sorghum (Sorghum vulgare Pers.) leaves infected with Drechslera sorghicola were investigated through five recognizable stages of disease development. Water-soaked yellowish brown spots developed two days after inoculation, turned brown with yellow halo, enlarged and coalesced at later stages of disease development. Healthy and infected leaves were analyzed for different biochemical constituents. The chlorophyll contents were decreased significantly with the progress of infection. The levels of reducing and total sugars increased while non-reducing sugars decreased to a significant extent with the progress of disease. The concentration of total phenolics, orthodihydroxy phenols, free and glycosidic phenols showed significant changes due to infection, whereas basic and acid phenols showed little or no change with disease development. Levels of phenolic compounds increased four days after inoculation and decrease thereafter, but the concentration was higher at every stage of disease development relative to healthy tissues. Polyphenol oxidase and peroxidase enzyme activities increased to varying degrees at different stages of infection. Analysis of protein fractions showed a significant increase with the progress of disease.

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Immunoelectron microscopic localization of partially purified antigens in adult Paragonimus iloktsuenensis

  • Lee, Ok-Ran;Chung, Pyung-Rim
    • Parasites, Hosts and Diseases
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    • v.39 no.2
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    • pp.119-132
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    • 2001
  • An immunoelectron microscopy employing immunogold labeling method was performed to detect tissue origin of Dl fraction (DIA) among 5 antigenic protein fractions partially purified by DEAE- anion exchange chromatography from water- soluble crude antigen (PIWA) of adult Paragonimus iloktsuenensis. Immune reactions of adult worm tissues with rabbit serum immunoglobulin immunized with crude antigen (PI-Ig) and D1 antigen (D1-Ig), as well as rat serum immunoglobulin infected with P. iloktsuenensis were observed. DlA showed strong antigenicity in the intestinal epithelium of the worms during the early infection period of 2-4 weeks after infection. The vitellaria also showed stronger antigenicity than the other tissue sites in immune reaction of tissues against all immunoglobulins from 4 to 33 weeks after vitelline development. Therefore, it is suggested that DlA was mainly originated from the intestinal epithelial tissues before the development of vitelline gland of the parasites. Immune-reactivity of two immunoglobulins (PI-Ig, Dl-Ig) was significantly different in intestinal epithelial cytoplasmic protrusions (CP) and intestinal epithelial secretory granules (SG). In the experimental group with Dl-Ig, gold particles were labeled significantly in CP than in SG when compared to the PI-Ig group. Thus, the major antigenic materials in Dl antigen having a strong antigenicity in the early infection period was considered to be originated from the intestinal epithelial tissue .

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