• Title/Summary/Keyword: protein chemistry

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Targeting Analysis of Lumenal Proteins of Chloroplast of Wheat using Proteomic Techniques

  • Kamal, Abu Hena Mostafa;Kim, Da-Eun;Oh, Myoung-Won;Chung, Keun-Yook;Cho, Yong-Gu;Kim, Hong-Sig;Song, Beom-Heon;Lee, Chul-Won;Uozumi, Nobuyuki;Choi, Jong-Soon;Cho, Kun;Woo, Sun-Hee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2010.05a
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    • pp.14-14
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    • 2010
  • Plastid proteomics are essential organelles present in virtually all cells in plants and green algae. Plastids are responsible for the synthesis and storage of key molecules required for the basic architecture and functions of plant cells. The proteome of plastid, and in particular of chloroplast, have received significant amounts of attention in recent years. Various fractionation and mass spectrometry (MS) techniques have been applied to catalogue the chloroplast proteome and its sub-organelles compartments. To better understanding the function of the lumenal sub-organelles within the thylakoid network, we have carried out a systematical analysis and identification of the lumenal proteins in the thylakoid of wheat by using Tricine-SDS-PAGE, and LTQ-ESI-FTICR mass spectrometry followed by SWISS-PROT database searching. We isolation and fractionation these membrane from fully developed wheat leaves using a combination of differential and gradient centrifugation couple to high speed ultra-centrifuge. After collecting all proteins to eliminate possible same proteins, we estimated that there are 407 different proteins including chloroplast, chloroplast stroma, lumenal, and thylakoid membrane proteins excluding 20 proteins, which were identified in nucleus, cytoplasm and mitochondria. A combination of these three programs (PSORT, TargetP, TMHMM, and TOPPRED) was found to provide a useful tool for evaluating chloroplast localization, transit peptide, transmembranes, and also could reveal possible alternative processing sites and dual targeting. Finally, we report also sub-cellular location specific protein interaction network using Cytoscape software, which provides further insight into the biochemical pathways of photosynthesis. The present work helps understanding photosynthesis process in wheat at the molecular level and provides a new overview of the biochemical machinery of the thylakoid in wheat.

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Effect of Sub- and Super-critical Water Treatment on Physicochemical Properties of Porcine Skin

  • Jo, Yeon-Ji;Kim, Jae-Hyeong;Jung, Kyung-Hun;Min, Sang-Gi;Chun, Ji-Yeon
    • Food Science of Animal Resources
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    • v.35 no.1
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    • pp.35-40
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    • 2015
  • Super- and sub-critical water treatments have been of interest as novel methods for protein hydrolysis. In the present study, we studied the effect of sub-critical water (Sub-$H_2O$, $300^{\circ}C$, 80 bar) treatment as well as super-critical water (Super-$H_2O$, $400^{\circ}C$, 280 bar) treatment on the physicochemical properties of porcine skin (PS), which has abundant collagen. Porcine skin was subjected to pre-thermal treatment by immersion in water at $70^{\circ}C$, and then treated with sub- or super-critical water. Physicochemical properties of the hydrolysates, such as molecular weight distribution, free amino acid content, amino acid profile, pH, color, and water content were determined. For the molecular weight distribution analysis, 1 kDa hydrolyzed porcine skin (H-PS) was produced by Super-$H_2O$ or Sub-$H_2O$ treatment. The free amino acid content was 57.18 mM and 30.13 mM after Sub-$H_2O$ and Super-$H_2O$ treatment, respectively. Determination of amino acid profile revealed that the content of Glu (22.5%) and Pro (30%) was higher after Super-$H_2O$ treatment than after Sub-$H_2O$ treatment, whereas the content of Gly (28%) and Ala (13.1%) was higher after Sub-$H_2O$ treatment. Super-$H_2O$ or Sub-$H_2O$ treatment affected the pH of PS, which changed from 7.29 (Raw) to 9.22 (after Sub-$H_2O$ treatment) and 9.49 (after Super-$H_2O$ treatment). Taken together, these results showed that Sub-$H_2O$ treatment was slightly more effective for hydrolysis than Super-$H_2O$ was. However, both Sub-$H_2O$ and Super-$H_2O$ treatments were effective processing methods for hydrolysis of PS collagen in a short time and can be regarded as a green chemistry technology.

Enzymatic Properties of Cytidine Deaminase Encoded by cdd Gene in Bacillus subtilis (Bacillus subtilis의 cdd 유전자에 의해 코드되는 Cytidine Deaminase의 효소학적 성질)

  • Song, Bang-Ho;Yoon, Mi-Sook;Kim, Kyung-Hwa;Yeo, Jeung-Sook;Jan Neuhard
    • Microbiology and Biotechnology Letters
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    • v.16 no.6
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    • pp.468-475
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    • 1988
  • The cloned B. subtilis cdd gene encoding cytidine/2'deoxycytidine deaminase (EC 3.5.4.5) was expressed in the cdd deficient B. subtilis mutant ED40. The gene was isolted from the cdd complementing plasmid pSO21, and inserted into the EcoR1/Pvu1 sites of pGB215-110 ΔB, which is a temperature sensitivie E. coli-B. subtilis shuttle vector. In the transformed B. subtilis ED4O harboring the resulting plasmid pSO100, cdd was expressed at several hundred fold elevated levels, and the cytidine deaminase activity in E. coli containing pSO100 was twice the level in B. subtilis/pSO0100. The Km value for cytidine of the partially purified enzyme is 1.88$\times$10$^{-4}$M at pH 7.0 and the V$_{max}$ = 11.1 $\mu$mol/min/mg of protein. The enzyme was completely inhibited by 0.1M mercaptoethanol and HgCl$_2$. The inhibition by p-chrolomercurybenzoic acid showed a Ki = 5 uM. These results suggest that sulfhydryl reagents block an active site thiol group, and/or disturb the formation of the tetrameic holoenzyme.

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Hemato-Biochemical Indices of Hanwoo Cattle Raised at Different Altitudes (서로 다른 고도에서 사육된 비육한우의 혈액-혈청 지수)

  • Ji, Joong-Ryong;Alam, Md. Rafiqul;Na, Chong-Sam;Kim, Nam-Soo
    • Journal of Veterinary Clinics
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    • v.28 no.2
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    • pp.219-224
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    • 2011
  • The health status of three groups of Hanwoo steers (n=157) aged 2-3 years, raised at three different altitudes (600 m, n=50; 200-400 m, n=58; plane land, n=49) and environment with more or less similar management have been evaluated through hematological, biochemical and globulin examinations in order to determine the optimum environment suitable for raising cattle while at the same time minimizing the risk of disease. Five mL of blood samples from each animal were collected by jugular veinipuncture and 2 mL was transferred to a tube containing EDTA for complete blood count (CBC) and 3 mL in lithium heparin for chemistry screening (CS) and immunoassay. Among the CBC parameters a significantly higher white blood cell count (tWBC), total red blood cell count (tRBC), hemoglobin (Hb) and packed cell volume (PCV) were noticed in the high altitude groups, whereas those were lower in the plane land group. In the CS parameters higher levels of aspartate aminotransferase (AST), creatinine, lactate dehydrogenase (LDH) and total bilirubin (TBL) were found in the plane land group, whereas those were lower in the high altitude groups. The total protein (significantly) and globulins were higher in the 600 m group. The results of this study revealed that the overall health status of the Hanwoo cattle based on the hemato-biochemical indices was superior in the highest altitude and inferior in the plane land group but all the parameters were within the reference range in all the groups. Therefore, for recommendation of a suitable environment at an appropriate altitude for raising cattle there need to be further studied along with the hemato-biochemical parameters; considering, breeding, feeding, management, marketing, waste disposal and other factors.

Effects of Storing Time on the Values of the Clinical Biochemistry in Sprague-Dawley(SD) Rats (랫드 혈청의 저장기간에 따른 혈액생화학치 변화)

  • Son, Hwa-Young;Lee, Hyun-Sook;Kim, Young-Hee;Kim, Yong-Beom;Kim, Il-Hwan;Ha, Chang-Su;Kang, Boo-Hyon
    • Korean Journal of Veterinary Pathology
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    • v.3 no.2
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    • pp.87-91
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    • 1999
  • The present study was undertaken to compare the variation on serum biochemical values by storage in the rats. Sera were prepared from 30 Sprague-Dawley rats of each sex. 5 aliquots from each serum were placed in a -80$^{\circ}C$ freezer with the exception of I aliquots which was analyzed immediately. The analysis was performed on the following months; 1, 2, 3, 6, and 12 months after freezing. The parameters measured) were aspartate aminotransferase(AST), alanine aminotransferase(ALT), alkaline phosphatase(ALP), blood urea nitrogen(BUN) creatinine(CRE), glucose(GLU), total cholesterol(TCHO), triglyceride (TG), total protein(TP), albumin(ALB), total bilirubin(TBIL), calcium(Ca$\^$++/), inorganic phosphorus(IP), creatine kinase (CK), phospholipid(PL), albumin-globulin ratio(A/G), sodium(Na$\^$+/), potassium(K$\^$+/), and chloride(Cl$\^$-/) The statistical analysis with Repeated Measures ANOVA, did not show statistical significance in the parameters of AST, ALT, BUN, TG, CK, A/G, Na$\^$+/ of 1 month freezed sera, in those of AST, TG, CK, K$\^$+/) of 2 month freezed sera, in those of AST, ALT, BUN, CRE, TCHO, TP, TBIL, CK, PL, Na$\^$+/), K$\^$+/), Ct on month fteezed sera, in those of Cl$\^$-/ of 6 month fteezed sera, and in those of ALT, TG, ALB of 12 month freezed sera in male SD rats. On the other hand, it did not show statistical significance in the parameters of AST, ALT, ALP, BUN, GLU, TCHO, TG, TBIL, CK, PL, A/G, Na$\^$+/ of 1 month freezed sera, in those of AST, TCHO of 2 month freezed sera, in those of AST, BUN, CRE, TCHO, TP, TBIL, CK, PL of 3 month freezed sera, in those of TCHO, IP, PL of 6 month freezed sera, and in those of ALB of 12 month freezed sera in female SD rats. On the basis of the results, although there are some statistical variations in the biochemical values of the sera, it is suggested that if sera are analysed at the same time before 12 months storage in a -80 $^{\circ}C$ freezer, the storing time does not affect the biochemical evaluation of the sera in SD rats.

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Serum Chemical Values of Korean Native Goats from Birth to Maturity (한국재래산양(韓國在來山羊)의 성장(成長)에 따르는 혈청화학치(血淸化學値)의 변동(變動))

  • Mum, Hi Cheol
    • Korean Journal of Veterinary Research
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    • v.15 no.2
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    • pp.187-198
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    • 1975
  • Although considerable research has been done on the blood chemistry of domestic animals, little work has been made of the changes associated with age. Moreover, the records about physiology of the goat were not much available in Korea, and a comprehensive survey of the blood values of the Korean native goat has not been made. The object of the present investigation was to make good this deficiency and to suggest standards for the blood chemical values of Korean native goats from birth to maturity. The goats were kept under average farming conditions in Korea. Blood samples were collected from the jugular vein at birth, at one and four days, at one, two, three and four weeks, and at two, three, six, nine and twelve months of age. The results obtained in this work were summarized as follows: 1. The highest concentration of serum glucose was observed at birth and it decreased gradually until three months of age, showing a steady state thereafter. 2. The concentration of total serum protein was a little higher at one day of age than at birth and showed a steady state until seven days of age; then it decreased slightly at two weeks of age and recovered at three weeks of age and showed a steady state until twelve months of age. The concentration of serum albumin was increased a little at one day of age and revealed a steady state thereafter. The concentration of serum globulin showed three phases - a fall during the first two weeks to reach minimum, a rise to the fourth week, and a fall at two months of age, showing a steady state thereafter. The lowest value of albumin fraction was observed at birth, then the value increased gradually until three months of age and revealed a stealer state thereafter. The highest value of globulin fraction was observed at birth, then the gradual decrease of the value was seen until three months of age; thereafter the value showed a steady state. 3. Albumin/globulin ratio was the minimum at birth, then it increased to reach a maximum at two weeks of age and decreased a little thereafter. 4. The concentration of total serum cholesterol showed a gradual increase during the first three months and fell to reach adult revel at six months of age. 5. Urea nitrogen in serum decreased during the first week to reach a minimum, then it increased at three months of age, and showed a fell to mature level at six mouths of age. 6. The concentration of serum creatinine was not affected by age. 7. The concentration of total serum calcium was a little higher during the first two weeks than the other period during the first year of life. 8. The concentration of serum inorganic phosphorus showed a gradual rise to reach a maximum and a gradual fall to reach adult level at nine months of age. 9. No significant sex differences of serum chemical values were recognized.

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Uptake, Assimilation and Translocation of Ammonium or Nitrate in Italian Ryegrass

  • Kim, Tae-Hwan;Lee, Bok-Rye;Jung, Woo-Jin;Kim, Dae-Hyun;Chung, Soon-Ju;Kim, Kil-Yong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.4
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    • pp.303-308
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    • 2001
  • To investigate the partitioning of newly absorbed N derived from NO$_3$- and NH$_4$$^{+}$, 6 mM $K^{15}$ NO$_3$ or 3 mM ($^{15}$ NH$_4$)$_2$ was fed continuously in Italian ryegrass (Lolium multiflrum L.) for 7 days. Nitrogen metabolites (nitrate, amino acid, soluble- and insoluble protein) were analyzed at the end of $^{15}$ N feeding. Dry weight in shoot, stubble and root was not significantly different between NO$_3$$^{[-10]}$ and NH$_4$$^{+}$ feeding. Total nitrogen content in all three organs was significantly higher in NH$_4$$^{+}$ than NO$_3$$^{[-10]}$ feeding. Sum on N content in reduced N fractions (amino acids + proteins) in shoot, stubble and roots in NH$_4$$^{+}$ feeding increased by 13.3, 12.5 and 35.4 %, respectively, compared to NO$_3$$^{[-10]}$ feeding. The Relative Specific Activity (RSA, percentage of newly absorbed $^{15}$ N relative to total N in a sample) values of amino acids and insoluble proteins were significantly higher in NH$_4$$^{+}$ feeding. Total amount of newly absorbed $^{15}$ N in NO$_3$$^{[-10]}$ and NO$_3$$^{[-10]}$ feeding was 52.3 and 69.5 mg/plant on dry matter basis, respectively. In both NH$_4$$^{+}$ and NO$_3$$^{[-10]}$ grown plants, most of the N was allocated to the shoot, 67.5% in NH$_4$$^{+}$ feeding and 58.8% NO$_3$$^{[-10]}$ feeding, respectively. The $^{15}$ N amount incorporated in the reduced N compounds (amino acids and proteins) in NH$_4$$^{+}$ grown plants significantly increased by 74.8% compared to NO$_3$$^{[-10]}$ grown plants. The increase of the $^{15}$ N amount assimilated to amino acids in NH$_4$$^{+}$ grown plants was remarkably higher in roots as more than 7.25 times compared to NO$_3$$^{[-10]}$ feeding. These results indicated that Italian ryegrass was much efficiently utilized NH$_4$$^{+}$-N for the synthesis of reduced N compounds.reduced N compounds.

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Preparation and Characterization of Urushiol Free Fermented Rhus verniciflua Stem Bark (FRVSB) Extracts (Urushiol이 제거된 발효옻 추출물의 제조와 특성)

  • Choi, Han-Seok;Yeo, Soo-Hwan;Jeong, Seok-Tae;Choi, Ji-Ho;Park, Hyo-Suk;Kim, Myung-Kon
    • Korean Journal of Food Science and Technology
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    • v.44 no.2
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    • pp.173-178
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    • 2012
  • The water extract of the fermented $Rhus$ $verniciflua$ stem bark (FRVSB) was prepared by hot water extracting at $100^{\circ}C$ for 8 h. The urushiol content of the FRVSB water extract was determined by HPLC. The urushiol was not contained in FRVSB water extract, whereas $Rhus$ $verniciflua$ stem bark (RVSB) water extract contained 3.4 mg%. At the lab scale size, suitable water extraction condition for a total solid, polyphenol and flavonoid from FRVSB was at over $100^{\circ}C$ for 6-8 h. The total solid contents was reduced in pilot scale processing system, with 5.7% of the extraction yield. The proximate composition (%) of FRVSB water extract obtained from industrial installation was moisture 4.34, crude fat 1.69, crude protein 10.21, and crude ash 15.80. Gallic acid (1,090.5 mg%) was the most abundant compound in phenolic acids, while fisetin (135.7 mg%) was the predominant flavonoid. The free sugar content was mannitol of 3.48%, glycerol of 0.19%, and glucose of 0.19%. Alanine (244 ppm), serine (231 ppm), and leucine (218 ppm) were predominant amino acids.

The Anti-Fibrogenic Effect of a Pharmaceutical Composition of[5-(2-Pyrazinyl)-4-methyl-1,2-dithiol-3-thione] (Oltipraz) and Dimethyl-4,4′-dimethoxy-5,6,5′,6′-dimethylene dioxybiphenyl-2,2′-dicarboxylate (DDB)

  • Kang, Keon-Wook;Kim, Yoon-Gyoon;Kim, Choon-Won;Kim, Sang-Geon
    • Archives of Pharmacal Research
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    • v.25 no.5
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    • pp.655-663
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    • 2002
  • Liver fibrosis is a prepathological state wherein damaged liver tissues in chronic liver diseases, such as hepatitis, are not repaired to normal tissues, but converted to fibrous tissue. 5-(2-Pyrazinyl)-4-methyl-1,2-dithiol-3-thione (oltipraz), a cancer chemopreventive agent, is effective against a wide variety of chemical carcinogens. Recently, we reported that oltipraz inhibits liver fibrogenesis (Kang et al., 2002). In the present study, the effects of oltipraz in combination with dimethyl-4,4'-dimethoxy-5,6,5',6'-dimethylene dioxybiphenyl-2,2'-dicarboxylate (DDb) on dimethylnitrosamine (DMN)-induced liver fibrogenesis were assessed in rats. Oltipraz (30 mg/kg body weight, po, 3 times per week for 4 weeks) was found to inhibit the increases in plasma ALT, AST and bilirubin by DMN, whereas DDB (30 mg/kg body weight, po, 3 times per week for 4 weeks) attenuated the increases in the plasma ALT and bilirubin. The lowered plasma protein and albumin contents in DMN-treated rats were completely restored by oltipraz, but not by DDB. DDB decreases liver cell injury and inflammation through inhibition of nuclear factor-kB. DMN increased the accumulation of liver collagen, as indicated by the increase in the 4-hydroxyproline content in liver homogenates, which was reduced by treatment with oltipraz, but not by DDB. Given the differential effect between oltipraz and DDB, the potential enhancement of antifibrotic efficacy by the drugs was assessed in the animal model. Despite the minimal effect of DDB on DMN-induced fibrogenesis, DDB (5-25 mg/kg), administered together with oltipraz (25-5 mg/kg), showed an additive protective effect against hepatotoxicity and fibrosis induced by DMN, which was shown by the blood chemistry parameters and histopathological analysis. The adequate composition ratio of oltipraz to DDB was 5:1. These results provide information on the pharmaceutical composition, comprising of oltipraz and DDB as the active components, for the treatment and/or prevention of liver fibrosis and cirrhosis.

Postharvest Changes in Quality and Biochemical Components of Leaf Lettuce (상치의 수확후(收穫後) 품질(品質) 및 성분변화(成分變化))

  • Kim, Seong Yeol;Hong, Young Pyo;Choi, Woo Young
    • Korean Journal of Agricultural Science
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    • v.12 no.1
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    • pp.128-138
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    • 1985
  • This study was conducted to elucidate the shelf-life and quality changes in relation to biochemical components in leaf lettuce (Lactuca sativa L.). The shelf-life of leaf lettuce at room temperature was 2 to 3 days. But it was extended to 3 weeks by packaging in a 0.01 mm thick polyethylene film sack when stored at $3^{\circ}C$. The ascorbic acid contents of fresh leaf lettuce was 25 mg per 100 gram fresh weight. The acid at room temperature was almost destroyed after 4 days storage. But the contents of ascorbic acid at $3^{\circ}C$ maintained about 50 to 60% of the initial level in packaging of polyethylene film sack after 8 days storage. The content of chlorophyll was greatly decreased at room temperature but no significant changes were found at $3^{\circ}C$. The changes of total sugar and reducing sugar contents during storage were not very different between treatments. The contents of alkali soluble protein and free amino acid gradually increased in the treatments of polyethylene film sack packaging during storage in general, but the contents decreased after the increase in control treatment. Nucleic acid content, peroxidase and polyphenoloxidase activities were measured and discussed in relation to the leaf senescence.

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