• Title/Summary/Keyword: cell wall protein

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Extraction and Purification of Antitumor Protein-bound Polysaccharides from Mycelia of Lentinus edodes (표고버섯 균사체로부터 항암 단백다당체의 추출 및 정제)

  • Park, Ki-Moon;Lee, Byung-Woo
    • Korean Journal of Food Science and Technology
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    • v.30 no.5
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    • pp.1236-1242
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    • 1998
  • Korean Lentinus edodes SR-1 was cultured to multiply the mycelia in the complete broth medium (C/N=13.1) for mushroom, and protein-bound polysaccharides were extracted from the cultured broth containing mycelia (The whole cultured broth was used to increase the yields: 80% of protein-bound polysaccharides were existed at the cell wall of mycelia and 20% of those were secreted extracellularily in this culture). Protein-bound polysaccharides in the cultured broth containing mycelia were extracted by using three different methods: 1) Extraction with hot water, 2) Disintegration of cell wall by glass bead mill treatment before extraction with hot water, and 3) Cellulase treatment before extraction with hot water. The highest yield was obtained (930 mg polysaccharides/100 mL culture broth) when protein-bound polysaccharides were extracted with 2) method. The extracted crude protein-bound polysaccharides were purified using protease, DEAE-cellulose and Sephadex G-100. The growth inhibition activity for $P_{388}$, mouse leukemic cell, increased (53.7, 62.2, 93.7% and 97.4%) as the purification level increased. Protein-bound polysaccharides contained 46.1% of polysaccharides, 7.3% of protein, and trace amounts of minerals. Polysaccharides contained glucose, galactose, xylose and mannose. The content of proline and glycine were high, however, methionine and leucine were not found. The major minerals were Na, K, Zn, and Ca.

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Recent Advances in the Studies of Self-Incompatibility of plants (식물의 자가불화합성, 최근의 진보)

  • 한창열;한지학
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.5
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    • pp.253-275
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    • 1994
  • Many flowering plants possess genetically controlled self -incompatibility (SI) system that prevents inbreeding and promotes outcrosses. SI is usually controlled by a single, multiallelic S-locus. In gametophytically controlled system, SI results when the S-allele of the pollen is matched by one of the two S-alleles in the style, while in the sporophytic system self-incompatible reaction occurs by the interaction between the pistil genotype and genotype of, not the pollen, but the pollen parent In the former system the self-incompatible phenotype of pollen is determined by the haploid genome of the pollen itself but in the latter the pollen phenotype is governed by the genotype of the pollen parent along with the occurrence of either to-dominant or dominant/recessive allelic interactions. In the sporophytic type the inhibition reaction occurs within minutes following pollen-stigma contact, the incompatible pollen grains usually failing to germinate, whereas in gametophytic system pollen tube inhibition takes place during growth in the transmitting tissue of the style. Recognition and rejection of self pollen are the result of interaction between the S-locus protein in the pistil and the pollen protein. In the gametophytic SI the S-associated glycoprotein which is similar to the fungal ribonuclease in structure and function are localized at the intercellular matrix in the transmitting tissue of the style, with the highest concentration in the collar of the stigma, while in the sporophytic SI deposit of abundant S-locus specific glycoprotein (SLSG).is detected in the cell wall of stigmatic papillae of the open flowers. In the gametophytic system S-gene is expressed mostly at the stigmatic collar the upper third of the style length and in the pollen after meiosis. On the other hand, in the sporophytic SI S-glycoprotein gene is expressed in the papillar cells of the stigma as well as in e sporophytic tape is cells of anther wall. Recognition and rejection of self pollen in the gametophytic type is the reaction between the ribonuclease in the transmitting tissue of the style and the protein in the cytoplasm of pollen tube, whereas in the sporophytic system the inhibition of selfed pollen is caused by the interaction between the Sycoprotein in the wall of stigmatic papillar cell and the tapetum-origin protein deposited on the outer wall of the pollen grain. The claim that the S-allele-associated proteins are involved in recognition and rejection of self pollen has been made merely based on indirect evidence. Recently it has been verified that inhibition of synthesis of S$_3$ protein in Petunia inflata plants of S$_2$S$_3$ genotype by the antisense S$_3$ gene resulted in failure of the transgenic plant to reject S$_3$ pollen and that expression of the transgenic encoding S$_3$ protein in the S$_1$S$_2$ genotype confers on the transgenic plant the ability to reject S$_3$ pollen. These finding Provide direct evidence that S-proteins control the s elf-incompatibility behavior of the pistil.

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Inactivation of the Wall-Associated De-N-acetylase (PgdA) of Listeria monocytogenes Results in Greater Susceptibility of the Cells to Induced Autolysis

  • Popowska, Magdalena;Kusio, Monika;Szymanska, Paulina;Markiewicz, Zdzislaw
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.932-945
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    • 2009
  • Several species of Gram-positive bacteria have cell wall peptidoglycan (syn. murein) in which not all of the sugar moieties are N-acetylated. This has recently been shown to be a secondary effect, caused by the action of a peptidoglycan N-acetylglucosamine deacetylase. We have found that the opportunistic pathogen Listeria monocytogenes is unusual in having three enzymes with such activity, two of which remain in the cytoplasm. Here, we examine the enzyme (PgdA) that crosses the cytoplasmic membrane and is localized in the cell wall. We purified a hexa-His-tagged form of PgdA to study its activity and constructed a mutant devoid of functional Lmo0415 (PgdA) protein. L. monocytogenes PgdA protein exhibited peptidoglycan N-acetylglucosamine deacetylase activity with natural substrates (peptidoglycan) from both L. monocytogenes and Escherichia coli as well as the peptidoglycan sugar chain component N-acetylglucosamine, but not with N-acetylmuramic acid. As was reported recently [6], inactivation of the structural gene was not lethal for L. monocytogenes nor did it affect growth rate or morphology of the cells. However, the pgdA mutant was more prone to autolysis induced by such agents as Triton X-100 and EDTA, and is more susceptible to the cationic antimicrobial peptides (CAMP) lysozyme and mutanolysin, using either peptidoglycan muramidases or autolysis-inducing agents. The pgdA mutant was also slightly more susceptible than the wild-type strain to the action of certain beta-lactam antibiotics. Our results indicate that protein PgdA plays a protective physiological role for listerial cells.

Changes in Soluble Proteins during Softening of Persimmon and Jujube Fruits (감과 대추의 연화중 가용성 단백질의 변화)

  • Seo, Chi-Hyeong;Shin, Seung-Ryeul;Jeung, Yong-Jin;Kim, Kwang-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.2
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    • pp.175-179
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    • 1997
  • Changes of protein contents and chromatogram patterns by gel filtration chromatography was investigated for the purpose of studying changes of Proteins during softening of persimmon and jujube fruits. Contents of water-soluble and salt-soluble proteins were increased during softening of persimmon and jujube fruits, but that of cell wall-bound proteins was decreased. After performing a gel filtration of water-soluble protein, one peak was separated in mature persimmon fruits and three peaks in soft persimmon fruits. In the case of jujube fruits, there were three peaks in both of mature and soft fruits. Pattern of salt-soluble and cell wall-bound proteins by gel filtration chromatography hardly changed during softening. During softening of two fruits, the contents of water- soluble and salt-soluble proteins appeared to be increased on the same fractions with the decreasing in content of cell wall-bound proteins.

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Identification and Expression Patterns of fvexpl1, an Expansin-Like Protein-Encoding Gene, Suggest an Auxiliary Role in the Stipe Morphogenesis of Flammulina velutipes

  • Huang, Qianhui;Han, Xing;Mukhtar, Irum;Gao, Lingling;Huang, Rongmei;Fu, Liping;Yan, Junjie;Tao, Yongxin;Chen, Bingzhi;Xie, Baogui
    • Journal of Microbiology and Biotechnology
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    • v.28 no.4
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    • pp.622-629
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    • 2018
  • Expansins are cell wall proteins that mediate cell wall loosening and promote specific tissue and organ morphogenesis in plants and in some microorganisms. Unlike plant expansins, the biological functions of fungal expansin-like proteins have rarely been discussed. In the present study, an expansin-like protein-encoding fvexpl1 gene, was identified from Flammulina velutipes by using local BLAST. It consisted of five exons with a total length of 822 bp. The deduced protein FVEXPL1 contained 274 amino acids with a predicted molecular mass and isoelectric point of 28,589 Da and pH 4.93, respectively. The first 19 amino acids from the N terminal are the signal peptide. Phylogenetic analysis and multiple protein alignment indicated FVEXPL1 was an expansin-like protein. The expression level of fvexpl1 gene in the stipe was significantly higher than that in the mycelia, primordia, and cap. However, the expression level of fvexpl1 gene was significantly higher in the fast elongation region of the stipe as compared with the slow elongation region. Expression analysis indicated that fvexpl1 gene might have an auxiliary role in the stipe morphogenesis of F. velutipes.

Changes of Peroxidsse Isozyme Pattern and Formation of Cell Wall of Hairy Root Irlduced by Agrohacterium rhisogenes from Potato Tuber (감자(Solanum tuberosum. L) 괴경에 있어서 Agrobacterium rhizogenes에 의해 형질전환된 조직세포의 세포벽 형성과 peroxidase isozyme 양상의 변화)

  • 정현숙;김영희
    • KSBB Journal
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    • v.6 no.1
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    • pp.27-33
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    • 1991
  • Hairy root induced by A. rhizogenes from potato tuber (Solanm tuberosum L.) synthesized the agropine and mannopine which were demonstrated with paper electrophoresis. And the starch contents in hairy root were increased gradually following the developmental stage. But protein contents were decreased. The activity of ${\beta}-glucan$ synthetase II(GSII) which is related to the cell wall biosynthesis was stimulated in hairy root on the developmental stage. And chloropromazine did not influence the activity of GS II while verapamil inhibited about 60% of the activity GS II. Therefore, these results showed $Ca^{2+}$ to be effective factor in the cell wall formation. Isozyme pattern of peroxidase was investigated in the callus and hairy root induced from potato tuber.

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Possible Roles of LAMMER Kinase Lkh1 in Fission Yeast by Comparative Proteome Analysis

  • Cho, Soo-Jin;Kim, Young-Hwan;Park, Hee-Moon;Shin, Kwang-Soo
    • Mycobiology
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    • v.38 no.2
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    • pp.108-112
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    • 2010
  • To investigate the possible roles of LAMMER kinase homologue, Lkh1, in Schizosaccharomyces pombe, whole proteins were extracted from wild type and lkh1-deletion mutant cells and subjected to polyacrylamide gel electrophoresis. Differentially expressed proteins were identified by tandem mass spectrometry (MS/MS) and were compared with a protein database. In whole-cell extracts, 10 proteins were up-regulated and 9 proteins were down-regulated in the mutant. In extracellular preparations, 6 proteins were up-regulated in the lkh1+ null mutant and 4 proteins successfully identified: glycolipid anchored surface precursor, $\beta$-glucosidase (Psu1), cell surface protein, glucan 1,3-$\beta$-glucosidase (Bgl2), and exo-1,3 $\beta$-glucanase (Exg1). These results suggest that Lkh1 is involved in regulating cell wall assembly.

Effects of NSP Degrading Enzyme on In vitro Digestion of Barley

  • Li, W.F.;Sun, J.Y.;Xu, Z.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.1
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    • pp.122-126
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    • 2004
  • A digestion trial in vitro was conducted to study effects of supplementation of NSP (non-starch polysaccharides) degrading enzyme (feed grade) on cell wall degradation and digestibility of nutrients in barley. The slices of barley were soaked in distilled water with or without 0.15% non-starch polysaccharides degrading enzyme. Microscopic examination of the slices showed that the endosperm cell wall of barley was completely degraded by the non-starch polysaccharides degrading enzyme. The residues and supernatant of digesta in vitro were separated by filtration with 0.1 mm nylon fabric. The residues were used for measurement of crude protein, crude fat, crude fiber, and moisture. The supernatant was used for determination of viscosity, as well as amino-nitrogen and glucose content. The results showed that compared with the control, the amino-nitrogen and glucose content of the supernatant increased by 17.58% (p<0.05) and 10.26% (p<0.05), respectively, while viscosity did not change. Enzyme supplementation increased the digestibilities of dry matter, crude protein, nitrogen-free extract, crude fat and crude fiber of barley by 18.1% (p<0.05), 20.3% (p<0.05), 16.4% (p<0.05), 26.9% (p<0.05) and 30.0% (p<0.05), respectively. The present study suggests that cell wall hydrolysis may contribute to improved nutrient digestion in vivo when non-starch polysaccharides degrading enzymes are fed to swine.

Determination of the Period of the Formation and Size of Sieve Element Area and Sieve Pore (Streptanthus tortus 조직배양 세포에서 사공의 형성시기와 사공 영역과 사공의 크기 결정)

  • Cho, Bong-Heuy
    • Journal of Plant Biotechnology
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    • v.29 no.1
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    • pp.41-44
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    • 2002
  • During the phloem development from parenchyma cells in a suspension culture of Streptanthus induced sucrose carrier and glucose carrier disappeared. Sieve element area and sieve pore induced suspension culture of Streptanthus were formed almost at the last period of the synthesis of sieve endoplasmic reticulum (SER) and p-protein. The new synthesized cell wall begann to digeste only after the new cell wall was surrounded by SER. The digested region of the cell wall and the formed region of sieve pore were regular comparatively. The completed sieve pore was an oval form, and the outer portion of sieve pore varied, ca 1.2 ${\mu}{\textrm}{m}$~1.6 ${\mu}{\textrm}{m}$ in longitudinal, 0.8 ${\mu}{\textrm}{m}$~1.3 ${\mu}{\textrm}{m}$ in tangential, and the inner size of sieve pore was irregular form of a star-like shape. The number of sieve pore between sieve cells was ca 2~7 per ${\mu}{\textrm}{m}$$^2$ and the sieve pore wall with callose was 0.05 ${\mu}{\textrm}{m}$~0.07 ${\mu}{\textrm}{m}$ in thickness. The energy for the formation of sieve element area and sieve pore might be supplied by mitochondria near the new cell wall and the role of SER remains to be illucidated.

Biological function of CpSlt2, an ortholog of the cell wall integrity (CWI) MAPK of Saccharomyces cerevisiae, in the chestnut blight fungus Cryphonectria parasitica

  • So, Kum-Kang;Ko, Yo-Han;Chun, Jeesun;Kim, Jung-Mi;Kim, Dae-Hyuk
    • 한국균학회소식:학술대회논문집
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    • 2018.05a
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    • pp.11-11
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    • 2018
  • Cryphonectria parasitica, chestnut blight fungus, has a characteristic of decreasing pathogenicity when infected with Cryphonectria hypovirus 1. C. parasitica is known to be one of the most representative model systems used to observe the interaction between viruses, plants and fungi. The mitogen-activated protein kinase (MAPK) pathway, which is well conserved in various organisms ranging from yeast to humans, functions in relaying phosphorylation-dependent signals within MAPK cascades to diverse cellular functions involved in the regulation of pheromone, cell wall integrity, and osmotolerance in filamentous fungi. Several genes in the MAPK pathway were revealed to be regulated by hypovirus, or to be involved in pathogenicity in C. parasitica. Among these pathways, the CWI pathway has aroused interest because CpBck1, an ortholog of yeast Bck1 (a CWI MAPKKK), was previously reported to be involved in cell wall integrity and sectorization. Interestingly, sporadic sectorization was observed in the CpBck1 mutant and sectored phenotypes were stably inherited in the progeny that were successively transferred from sectored mycelia. In this study, we analyzed the biological function of CpSlt2, downstream gene of CpBck1, to confirm whether the sectorization phenomenon occurred in the specific single gene or cell wall integrity (CWI) pathway. As results, the CpSlt2-null mutant exhibited marked changes in colonial growth, near absence of conidiation and aerial hyphae, abnormal pigmentation, CWI-related phenotypic defects, and dramatically impaired virulence. As cultivation of the mutant strains progressed, the majority of the colonies showed sporadic sectorization and mycelia from the sectored area stably maintained the sectored phenotype. These results suggest that the unique sectorization is CWI pathway-specific, though the components in the same CWI pathway have common and specific functions.

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