• Title/Summary/Keyword: cell wall degradation enzyme activity

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Changes in Cell Wall Components and Cell Wall-degrading enzymes during Softening of Fruits (과실의 연화중에 세포벽 성분과 세포벽분해효소의 변화)

  • 신승렬;김광수
    • Food Science and Preservation
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    • v.3 no.1
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    • pp.93-104
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    • 1996
  • The cell wall components of fruit include cellulose. hemicellulose, pectin, glycoprotein etc., and the cell wall composition differs according to the kind of fruit. Fruit softening occurs as a result of a change in the cell wall polysaccharides : the middle lamella which links primary cell walls is composed of pectin. and primary cell walls are decomposed by a solution of middle lamella caused due to a result of pectin degradation by pectin degrading enzymes during ripening and softening, During fruit ripening and softening, contents of arabinose and galactose among non-cellulosic neutral sugars are notably decreased, and this occurs as a result of the degradation of pectin during fruit repening and softening since they are side-chained with pectin in the form of arabinogalactan and galactan Enzymes involved in the degradation of the cell wall include polygalacturonase, cellulose, pectinmethylesterase, glycosidase, etc., and various studies have been done on the change in enzyme activities during the ripening and softning of fruit. Among cell wall-degrading enzymes, polygalacturonase has the greatest effect on fruit softening, and its activity Increases during the maturating and softening of fruit. This softening leads to the textural change of fruit as a result of the degradation of cell wall polysaccharides by a cell wall degrading enzyme which exists in fruit.

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Ripening Related Changes in Hot Pepper Fruit Cell Walls Structural Alterations of Cell Wall Polysaccharides and Separation of Galactosidase Isozymes (고추의 성숙에 따른 세포벽 다당류의 변화와 ${\beta}-Galactosidase$ Isozymes의 분리)

  • Kim, Soon-Dong;Kang, Meung-Su;Kim, Kwang-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.2
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    • pp.157-163
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    • 1985
  • Various cell wall polysaccharides and related enzyme activities in hot pepper fruit were determined at different stages of maturity. The uronic acid content of cell walls decreased between immature green and turning stage fruit and then increased by red ripe stage. In contrast, cellulose content of cell walls changed only a little during ripening. Total neutal sugar content of cell wall material decreased 50% and galactose content of the walls decreased about 80% by the turning stage. Polygalacturonase and ${\beta}-galactosidase$ activities, as well as total hemicellulose from isolated cell walls of ripening hot pepper fruit were studied using gel filtration chromatography. Polygalacturonase activity was not detectable but 5 isozymes of ${\beta}-galactosidase$ were resolved. The activities of the enzymes were relatively high and gel filtration showed that they differed in molecular weight. Hemicellulose content decreased during ripening and softening. The molecular weight profiles shifted from high molecular weight to low molecular weight polymers during ripening. The changes in cell walls that may be associated with fruit softening involve the alteration of hemicellulose prior to the degradation of wall-bound uronic acid. It is suggested that the decrease in cell wall galactose involved changes in turnover of new cell wall components.

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Identification of Sugar-Responsive Genes and Discovery of the New Functions in Plant Cell Wall

  • Lee, Eun-Jeong
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2007.04a
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    • pp.65-73
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    • 2007
  • The objective of this study is to understand how regulatory mechanisms respond to sugar status for more efficient carbon utilization and source-sink regulation in plants. So, we need to identify and characterize many components of sugar-response pathways for a better understanding of sugar responses. For this end, genes responding change of sugar status were screened using Arabidpsis cDNA arrays, and confirmed thirty-six genes to be regulated by sucrose supply in detached leaves by RNA blot analysis. Eleven of them encoding proteins for amino acid metabolism and carbohydrate metabolism were repressed by sugars. The remaining genes induced by sugar supply were for protein synthesis including ribosomal proteins and elongation factors. Among them, I focused on three hydrolase genes encoding putative $\beta$-galactosidase, $\beta$-xylosidase, and $\beta$-glucosidase that were transcriptionally induced in sugar starvation. Homology search indicated that these enzymes were involved in hydrolysis of cell wall polysaccharides. In addition to my results, recent transcriptome analysis suggested multiple genes for cell wall degradation were induced by sugar starvation. Thus, I hypothesized that enzyme for cell wall degradation were synthesized and secreted to hydrolyze cell wall polysaccharides producing carbon source under sugar-starved conditions. In fact, the enzymatic activities of these three enzymes increased in culture medium of Arabidopsis suspension cells under sugar starvation. The $\beta$-galactosidase encoded by At5g56870 was identified as a secretory protein in culture medium of suspension cells by mass spectrometry analysis. This protein was specifically detected under sugar-starved condition with a specific antibody. Induction of these genes was repressed in suspension cells grown with galactose, xylose and glucose as well as with sucrose. In planta, expression of the genes and protein accumulation were detected when photosynthesis was inhibited. Glycosyl hydrolase activity against galactan also increased during sugar starvation. Further, contents of cell wall polysaccharides especially pectin and hemicellulose were markedly decreased associating with sugar starvation in detached leaves. The amount of monosaccharide in pectin and hemicellulose in detached leaves decreased in response to sugar starvation. These results supported my idea that cell wall has one of function to supply carbon source in addition to determination of cell shape and physical support of plant bodies.

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Molecular Identification, Enzyme Assay, and Metabolic Profiling of Trichoderma spp.

  • Bae, Soo-Jung;Park, Young-Hwan;Bae, Hyeun-Jong;Jeon, Junhyun;Bae, Hanhong
    • Journal of Microbiology and Biotechnology
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    • v.27 no.6
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    • pp.1157-1162
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    • 2017
  • The goal of this study was to identify and characterize selected Trichoderma isolates by metabolic profiling and enzyme assay for evaluation of their potential as biocontrol agents against plant pathogens. Trichoderma isolates were obtained from the Rural Development Administration Genebank Information Center (Wanju, Republic of Korea). Eleven Trichoderma isolates were re-identified using ribosomal DNA internal transcribed spacer (ITS) regions. ITS sequence results showed new identification of Trichoderma isolates. In addition, metabolic profiling of the ethyl acetate extracts of the liquid cultures of five Trichoderma isolates that showed the best anti-Phytophthora activities was conducted using gas chromatography-mass spectrometry. Metabolic profiling revealed that Trichoderma isolates shared common metabolites with well-known antifungal activities. Enzyme assays indicated strong cell wall-degrading enzyme activities of Trichoderma isolates. Overall, our results indicated that the selected Trichoderma isolates have great potential for use as biocontrol agents against plant pathogens.

Effects of benzyladenine on the cell wall regeneration of soybean(Glycine max) protoplasts (대두(Glycine max) protoplast의 세포벽재생에 대한 benzyladenine의 영향)

  • Riu, Key-Zung;Park, Chang-Kyu
    • Applied Biological Chemistry
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    • v.35 no.6
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    • pp.507-512
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    • 1992
  • A ${\beta}-1,3-glucanase$ of soybean (Glycine max) was isolated, and the effects of benzyladenine(BA) on celluar levels of the enzyme content and activity were studied. The effects of BA on callose content in cell wall and wall regeneration of protoplasts were also studied to show promoting effect of cytokinin in cell wall regeneration and to elucidate action mode of cytokinin. The polypeptide of 21 kD was identified as ${\beta}-1,3-glucanase$, and the cellular content and activity of this polypeptide were decreased by BA treatment. The callose content in cell wall of callus and the wall regeneration of protoplasts were increased by BA treatment. These results indicate that cytokinin promotes cell wall regeneration by inhibition of callose degradation via decreasing ${\beta}-1,3-glucanase$ level in cell.

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Preliminary Studies on the Relationship between Reed and Bacterial Communities in the Salt Marsh Environment of Namyang Bay, Korea

  • Kwon, Kae-Kyoung;Je, Jong-Geel
    • Ocean and Polar Research
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    • v.24 no.1
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    • pp.47-53
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    • 2002
  • To evaluate the effect of reed population on the distribution and activities of microorganisms, vertical distribution of heterotrophic bacteria, degradation rate of cellulose, extracellular aminopeptidase activity (APA) and metabolic diversity based on GN2 Microlog plate were measured at two salt marsh stations in Hogok-ri, Namyang Bay, west coast of Korea. The number of heterotrophic bacteria at station 1 (reed population inhabited area) showed 2 to 6 times higher than that of station 2 (exposed area) with exception in the surface layer. Cellulose degradation rates in station 1 showed more than 50%. month-I and higher than that of station 2 (10.2 to 38.4%. $month^{-1}$). Yet the APA at two stations did not show difference except surface layer and suggested that APA might not be a significant factor in degrading marsh plant debris. Lipid class compounds, cell wall polymers and L-alanine were widely used by microorganisms. The number and activities of bacterial populations especially concerned in plant debris degradation seemed to be stimulated by the reed communities.

Isolation, Identification and Enzyme Properties of a Bacterium producing Alkaline Protease (Alkaline protease를 생산하는 미생물의 분리, 동정 및 효소성질)

  • Shin, Kong-Sik;Kang, Sang-Mo;Ko, Jung-Youn
    • Applied Biological Chemistry
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    • v.43 no.3
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    • pp.169-173
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    • 2000
  • For the development of enzyme detergent capable of effectively washing at low temperature, a bacterium producing alkaline protease was isolated from soil samples, and properties of the enzyme were investigated. The selected strain was Gram negative, rod shape$(0.6{\sim}0.7{\times}1.3{\sim}2.6\;{\mu}m\;in\;size)$ and motile. It had the degradation activity of aesculin, gelatin and casein, and was catalase-positive. The cell wall components was meso-DAP, and G+C mole contents was 43.3%. From these results, the strain was identified as Acinetobacter sp. KN-27. The activity of alkaline protease by this strain peaked with 3,300 D.U/mL after 36 hours in the liquid culture at $40^{\circ}C$. The optimal pH and temperature of the enzyme were pH 9 and $60^{\circ}C$, respectively. Alkaline protease produced by Acinetobacter sp. KN-27 has shown two active bands on the electrophoresis of native gel.

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Biodegradation of Mixture of Benzoate and m-Toluate with Pseudomonas sp. (Pseudomonas sp. 의한 Benzoate와 m-Toluate 혼합물의 생분해)

  • 정준영;김교창;조재민
    • Microbiology and Biotechnology Letters
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    • v.26 no.4
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    • pp.352-357
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    • 1998
  • Biodegradation of benzoate and m-toluate was investigated using a Pseudomonas sp. isolated in a continuous culture for 45 days with a step-wise increase of the subsrates. The optimum mixture ratio of benzoate and m-toluate was 75% and 25%, respectively. During 45-day culture, removal of benzoate and m-toluate, which was replaced 2,000 ppm on the 30th day were 94% and 79%, respectively, when COD removal rate was 80%. The enzymatic activity of catechol 1,2-dioxygenase increased and that of catechol 2,3-dioxygenase decreased as the concentration of m-toluate was increased. These results suggested that m-toluate induced enzyme activity for degradation of benzoate. The shape of isolated strain in the continuous culture was investigated with SEM and the results showed that the cell shape was more damage according to the higher concentration of aromatic hydrocarbons. Therefore, we suggested that the tolerance against aromatic hydrocarbons was related to not only enzymatic activity but also characteristic of cell membrane or cell wall.

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Cloning and Heterologous Expression of Acetyl Xylan Esterase from Aspergillus ficuum

  • Jeong, Hye-Jong;Park, Seung-Mun;Yang, Mun-Sik;Kim, Dae-Hyeok
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.153-156
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    • 2000
  • Xylan, the major hemicellulose component of many plants, occurs naturally in a partially acetylated form and lignin, the most resistant component in plant cell wall degradation, is also attached to ${\beta}-1,4-linked-D-xylose$ backbone through the ester linkage. Esterases are required to release the esterified substituent and acetyl esterases are important in the complete degradation of acetylated polysaccharides, like pectins and xylans. The gene(Axe) encoding acetyl xylan estarase(AXE) was isolated from genomic ${\lambda}$ library from Aspergillus ficuum. Nucleotide sequencing of the Axe gene indicated that the gene was separated with two intervening sequences and the amino acid sequence comparison revealed that it was closely related to that from A. awamori with the 92 % indentity. Heterologous expression of AXE was conducted by using YEp352 and Saccharomyces cerevisae 2805 as a vector and host expression system, respectively. The Axe gene was placed between GAL1 promoter and GAL7 terminator and then this recombinant vector was used to transform S. cerevisiae 2805 strain. Culture filtrate of the transformed yeast was assayed for the presence of AXE activity by spectrophotometry and, comparing with the host strain, four to five times of enzyme activity was detected in culture filtrate of transformed yeast.

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Adenine attenuates lipopolysaccharide-induced inflammatory reactions

  • Silwal, Prashanta;Lim, Kyu;Heo, Jun-Young;Park, Jong IL;Namgung, Uk;Park, Seung-Kiel
    • The Korean Journal of Physiology and Pharmacology
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    • v.22 no.4
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    • pp.379-389
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    • 2018
  • A nucleobase adenine is a fundamental component of nucleic acids and adenine nucleotides. Various biological roles of adenine have been discovered. It is not produced from degradation of adenine nucleotides in mammals but produced mainly during polyamine synthesis by dividing cells. Anti-inflammatory roles of adenine have been supported in IgE-mediated allergic reactions, immunological functions of lymphocytes and dextran sodium sulfate-induced colitis. However adenine effects on Toll-like receptor 4 (TLR4)-mediated inflammation by lipopolysaccharide (LPS), a cell wall component of Gram negative bacteria, is not examined. Here we investigated anti-inflammatory roles of adenine in LPS-stimulated immune cells, including a macrophage cell line RAW264.7 and bone marrow derived mast cells (BMMCs) and peritoneal cells in mice. In RAW264.7 cells stimulated with LPS, adenine inhibited production of pro-inflammatory cytokines $TNF-{\alpha}$ and IL-6 and inflammatory lipid mediators, prostaglandin $E_2$ and leukotriene $B_4$. Adenine impeded signaling pathways eliciting production of these inflammatory mediators. It suppressed $I{\kappa}B$ phosphorylation, nuclear translocation of nuclear factor ${\kappa}B$ ($NF-{\kappa}B$), phosphorylation of Akt and mitogen activated protein kinases (MAPKs) JNK and ERK. Although adenine raised cellular AMP which could activate AMP-dependent protein kinase (AMPK), the enzyme activity was not enhanced. In BMMCs, adenine inhibited the LPS-induced production of $TNF-{\alpha}$, IL-6 and IL-13 and also hindered phosphorylation of $NF-{\kappa}B$ and Akt. In peritoneal cavity, adenine suppressed the LPS-induced production of $TNF-{\alpha}$ and IL-6 by peritoneal cells in mice. These results show that adenine attenuates the LPS-induced inflammatory reactions.