• Title/Summary/Keyword: isolation wall

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The Types of Linkage of Carbohydrates in Wood Cell Wall (I) - The Isolation of Carbohydrates - (목재(木材) 세포벽중(細胞壁中)의 탄수화합물(炭水化合物) 간(間)의 결합(結合) 양식(樣式)(I) -탄수화합물(炭水化合物)의 단리(單離)-)

  • Lee, Sang-Pill;Lee, Jong-Yoon
    • Journal of the Korean Wood Science and Technology
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    • v.15 no.3
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    • pp.34-43
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    • 1987
  • This study was performed to find out the types of linkage of carbohydrates in wood cell walls. To study the structure of linkage of carbohydrates in wood cell walls, we have attempted to find out the method holocellulose preparation and optimum condition of enzyme hydrolysis in holocellulose, and fractionate oligosaccharide with products that hydrolized partly by acetolysis and deacetylation in holocellulose. We have achieved four results. These results as follow; 1. At first. we reacted in wood meal $NaClO_2$ 1g per lignin lg for one hour and then the same of quantity $NaClO_2$ for four hours. Through these experiments, we have developed new holocellulose preparation method which had low loss of carbohydrates and high effect of the delignification. 2. The optimum condition of enzyme hydrolysis of holocellulose which had lignin was 0.005M sodium acetate buffer (pH 5.0). We have achieved 7.2% reducing sugar through the procedure that reactioned 0.01g holocellulose putting enzyme 0.03g for 72 hours. It may be supposed that 5.5% of lignin contained in holocellulose prevented enzyme contaction from holocellulose and so this lignin has resulted in the low efficiency of enzyme hydrolysis. 3. We did not fractionated from oligosaccharides which were preparated by the method of acetolysis and deacetylation in holocellulose. The reason is that holocellulose having a lot of lignin prevented prefectly partial hydrolysis from the method of acetolysis and deacetylation. 4. We attempted analysis of six standard substances through HPLC apparatus having sugar pak 1 column which we have changed flow rate and the column temperature variably. These six standard substances were D-glucose, D-mannose, D-xylose, D-galactose and L-rhamnose, L-arabinose, But sugar pak 1 column was not fitted analysis of four substances because D-galactose, D-mannose, D-xylose, L-rhamnose were agreement with elution time. And so, we could not analize four standard substances with sugar pak 1 column.

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Isolation and Properties of a Protein, RCG-2, Having Chitinase, ${\beta}-1,3-Glucanase$ and Lysozyme Activities from Rice Leaves (Chitinase, ${\beta}-1,3-glucanase$ 및 lysozyme 효소활성을 보유한 벼잎 산성단백질 RCG-2)

  • Um, Sung-Yon;Kim, Su-Il
    • Applied Biological Chemistry
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    • v.37 no.1
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    • pp.49-55
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    • 1994
  • An acidic protein, RCG-2, containing chitinase and ${\beta}-1,3-glucanase$ activity conccurrently was purified from rice leaves by chromatofocusing and gel slicing. The purified enzyme gave a single band on polyacrylamide gel electrophoresis and its molecular weight was appeared to be 29.7 kd using SDS-PAGE. This enzyme also had lysozyme activity. The optimal temperature for both enzyme activities was $40^{\circ}C$, optimal pH were 4.0 for chitinase activity and 7.0 for ${\beta}-1,3-glucanase$ activity. $K_M$ and $V_{max}$ values for chitinase were 7.86 mM and $0.025\;{\mu}M/min.$, and those for ${\beta}-1,3-glucanase$ were 5.95 mM and $0.16\;{\mu}M/min.$ respectively. TLC analysis of the enzyme hydrolysates of chitooligosaccharides indicated that this enzyme acts as endochitinase.

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An Annealing Control Primer (ACP) System Used for the Isolation and Identification of Copper-Induced Genes in Alfalfa Leaves

  • Lee, Ki-Won;Lee, Sang-Hoon;Kim, Ki-Yong;Ji, Hee Chung;Park, Hyung Soo;Hwang, Tae Young;Choi, Gi Jun;Rahman, Md. Atikur
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.36 no.3
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    • pp.237-242
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    • 2016
  • Copper (Cu) is a necessary microelement for plants. However, high concentrations of Cu are toxic to plants that change the regulation of several stress-induced proteins. In this study, an annealing control primer (ACP) based approach was used to identify differentially expressed Cu-induced genes in alfalfa leaves. Two-week-old alfalfa plants (Medicago sativa L.) were exposed to Cu for 6 h. Total RNAs were isolated from treated and control leaves followed by ACP-based PCR technique. Using GeneFishing ACPs, we obtained several genes those expression levels were induced by Cu. Finally, we identified several genes including UDP-glucuronic acid decarboxylase, transmembrane protein, small heat shock protein, C-type cytochrome biogenesis protein, mitochondrial 2-oxoglutarate, and trans-2,3-enoyl-CoA reductase in alfalfa leaves. These identified genes have putative functions in cellular processes such as cell wall structural rearrangements, transduction, stress tolerance, heme transport, carbon and nitrogen assimilation, and lipid biosynthesis. Response of Cu-induced genes and their identification in alfalfa would be useful for molecular breeding to improve alfalfa with tolerance to heavy metals.

Screening and Isolation of Antagonistic Actinomyces #120 against the Kiwi Fruit Rot for the Environment-Friendly Culture of Kiwifruits (참다래의 친환경재배를 위한 과숙썩음병원균에 대한 길항성 방선균 #120의 선발 및 분리)

  • Cho, Jung-Il;Cho, Ja-Yong;Park, Yong-Seo;Son, Dong-Mo;Heo, Buk-Gu;Kim, Chul-Soo
    • Journal of Bio-Environment Control
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    • v.16 no.3
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    • pp.252-257
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    • 2007
  • This study was carried out to clarify the effects of antifungal Streptomyces sp. isolated from the soil grown kiwifruit on the growth inhibition of fruit rot (Botryosphaeria dothidea) infected in kiwi fruit plants in the southwestern districts of Jeonnam. Two hundred and fifty microorganisms were isolated and examined into the antifungal activity against Botryosphaeria dothidea. We screened and isolated six bacterial strains which have a strong inhibition against Botryosphaeria dothidea. And the best antifungal strain designated as the strain #120 showing 96.0% antifungal activity against Botryosphaeria dothidea was finally selected. The strain #120 was identified as Streptomyces sp. #120 based on its morphological, physiological, biochemical and chemotaxonomic characteristics.

Isolation of Streptomyces sp. KK565 as a Producer of ${\beta}-Amyloid$ Aggregation Inhibitor

  • Hwang, Sung-Eun;Im, Hyung-Min;Kim, Dong-Hoon;Shin, Hyun-Ju;Shin, Dong-Hoon;Park, Jeong-Eun;Jo, In-Ho;Kim, Chang-Jin;Yoo, Jong-Shin;Kang, Jong-Min;Lim, Dong-Yeon;Ahn-Jo, Snag-Mee;Kwon, Ho-Jeong
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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    • pp.809-814
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    • 2003
  • ${\beta}-amyloid$ ($A{\beta}$) peptides from the proteolytic processing of ${\beta}-amyloid$ precursor protein (${\beta}-APP$) aggregates in the brain to form senile plaques, and their aggregation plays a key role in pathogenesis of Alzheimer's disease (AD). To isolate an active compound that has an $A{\beta}$ aggregation-inhibitory activity, 2,000 microbial metabolite libraries were screened based on their ability to inhibit $A{\beta}$ aggregation by using both Congo red and thioflavin T assays. As a result, a water-soluble fraction of a soil microorganism, KK565, showed a potent $A{\beta}$ aggregation-inhibitory activity. The strain was identified as Streptomyces species, based on the cultural and morphological characteristics, the presence of diaminopimelic acid in the cell wall, and the sugar patterns for the whole-cell extract. In addition, the purification of active principle resulted in identifying a heat-unstable protein responsible for the $A{\beta}$ aggregation-inhibitory activity.

Isolation and Characterization of an Alkalophilic Cellulolytic Bacterium Pseudomonas sp. (호알칼리성 섬유소분해세균 Pseudomonas sp.의 분리 및 특성)

  • Lim, Sang-Ho;Yoon, Min-Ho;Choi, Woo-Young
    • Korean Journal of Agricultural Science
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    • v.25 no.1
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    • pp.124-130
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    • 1998
  • An alkalophilic bacterium, the strain AC-711 as a potent producer of alkaline cellulase, was selected among many isolates from soil environments. Morphological, physiological and chemical characteristics of the strain AC-711 suggested that it belongs to the genus Pseudomonas according to the Bergey's Manual of Systematic Bacteriology, however the G+C mol% (54.43) of its chromosomal DNA is lower than the normal values of the genus. The major cell wall fatty acids were determined as 15:0 and 17:0 anteiso. The production of alkaline CMCase by the strain was maximal when grown on the mediun containing 1% carboxymethyl cellulose, 0.1% $KH_2PO_4$, 0.02% $CoCl_2$, 0.02% Tween 80, 0.5% $Na_2CO_3$, 0.8% yeast extract, pH 10.3 at $30^{\circ}C$ for 3 days, and the most of enzyme was excreted into culture mediun.

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Physiological Studies on the Formation of Hairy Root by the A. rhizogenes. III. Attachment of A. rhizogenes strain A4 to Carrot(Daucus carota L.) Cells (Agrobacterium rhizogense에 Hairy Root 형성에 대한 생리학적 연구. III. 당근 세포에의 A. rhizogenes의 부착)

  • Hwang, B.;Hwang, S. J.;Ann, J. C.;Jo, H. S.
    • KSBB Journal
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    • v.4 no.2
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    • pp.94-98
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    • 1989
  • In vitro attachment experiments of bacteria to surface of host plant cell were carried out using C14 labeled cells of A. rhizogenes strain A4 and carrot protoplasts isolated from suspension culture of cells. Protoplasts were cocultivated with A. rhizogenes at various times after their isolation. Attachment kinetics showed that adherence of bacteria to protoplasts attained a maximum level within 120mins of co-cultivation. Maximum attachment occured at pH 6.0 and 24-35$^{\circ}C$. Bacterial attachment was observed at botg carrot cells with and without primary cell wall. The inhibition of transformation on the carrot root discs by A. rhizogenes was observed when non-related strain and heat inactivated bacterial strain cells were pretreated.

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Isolation and Analysis of the Yeast Mutant Gene, soo1-1, which Confers the Defect in $\beta$-1,3-glucan Biosynthesis (효모의 베타-1,3-글루칸 생합성 결함을 초래하는 돌연변이 유전자(soo1-1)의 분리 및 분석)

  • 이재준;이동원;김기현;박희문
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.42-48
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    • 2001
  • Allele rescue and sequence analysis of soo1-1 allele in Saccharomyces cerevisiae mutant LP0353 revealed that soo1-1 is identical to the previously reported ret1-1 allele, which has a base substitution of A for $G^{681}$ leading to an amino acid substitution of aspartic acid for $glycine^{227}$ in Soolp. However, it was revealed that the addition of osmotic stabilizer, such as 1.2M sorbitol can rescue the temperature sensitive phenotype of the ret1-1 mutant and that the soo1-1/ret1-1 mutation may confer defects in post-translational modification of proteins involved in the yeast cell wall biogenesis. Evidence for a putative role of 5th WD40 domain of the Soo1p/$\alpha$-COP in the construction and maintenance of cell walls was also presented by complementation test with deletion constructs of the SOOl.

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Isolation of Amylolytic Bifidobacterium sp. Int-57 and Characterization of Amylase

  • Ji, Geun-Eog;Han, Hee-Kyung;Yun, Seong-Wook
    • Journal of Microbiology and Biotechnology
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    • v.2 no.2
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    • pp.85-91
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    • 1992
  • The intestinal microflora of humans is an extraordinarily complex mixture of microorganisms, the majority of which are anaerobic microorganisms. The distribution of amylolytic microorganisms in the human large intestinal tract was investigated in various individuals of differing ages using anaerobic culture techniques. A large percentage of the amylolytic microorganisms present belonged to the Genus Bifidobacteria. The number of Bifidobacteria increased significantly at two years of age. Adults and children above 2 years old carried about $0.8{\times}10^9-2.0{\times}10^{10}$ colony forming units (CFU/gram) of amylolytic Bifidobacteria. Among these amylolytic Bifidobacteria, Int-57 was chosen for further studies. Between 65% and 85% of the amylase produced was secreted and the remaining amylase was bound to the cell wall facing the outside. Amylase production could be induced by starch in a stable form. When cells were grown on maltose or glucose, amylase production was much lower than on starch and amylase activity disappeared after 24 hours growth on these media. Partially purified enzymes showed optimum activity at a temperature of $50^{\circ}C$ and at an optimum pH of 5.5, respectively. Heat treatment at $70^{\circ}C$ for 30 minutes almost completely inactivated amylase. The hydrolysis products of starch were mainly maltose and maltotriose. Soluble starch, amylose, amylopectin, and $\gamma$-cyclodextrin($\gamma$-CD) were easily hydrolyzed. The rate of hydrolysis of $\alpha$-CD and $\beta$-CD was slower than that of $\gamma$-CD. Carboxymethyl cellulose, $\beta$-1, 3-glucan and inulin were not hydrolyzed.

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Isolation of Bacteria Associated with the King Oyster Mushroom, Pleurotus eryngii

  • Lim, Yun-Jung;Ryu, Jae-San;Shi, Shanliang;Noh, Won;Kim, Eon-Mi;Le, Quy Yang;Lee, Hyun-Sook;Ro, Hyeon-Su
    • Mycobiology
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    • v.36 no.1
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    • pp.13-18
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    • 2008
  • Eight distinct bacteria were isolated form diseased mycelia of the edible mushroom, Pleurotus eryngii. 16S rDNA sequence analysis showed that the isolates belonged to a variety of bacterial genera including Bacillus (LBS5), Enterobacter (LBS1), Sphingomonas (LBS8 and LBS10), Staphylococcus (LBS3, LBS4 and LBS9) and Moraxella (LBS6). Among them, 4 bacterial isolates including LBS1, LBS4, LBS5, and LBS9 evidenced growth inhibitory activity on the mushroom mycelia. The inhibitory activity on the growth of the mushroom fruiting bodies was evaluated by the treatment of the bacterial culture broth or the heat-treated cell-free supernatant of the broth. The treatment of the culture broths or the cell-free supernatants of LBS4 or LBS9 completely inhibited the formation of the fruiting body, thereby suggesting that the inhibitory agent is a heat-stable compound. In the case of LBS5, only the bacterial cell-containing culture broth was capable of inhibiting the formation of the fruiting body, whereas the cell-free supernatant did not, which suggests that an inhibitory agent generated by LBS5 is a protein or a heat-labile chemical compound, potentially a fungal cell wall-degrading enzyme. The culture broth of LBS1 was not inhibitory. However, its cell-free supernatant was capable of inhibiting the formation of fruiting bodies. This indicates that LBS1 may produce an inhibitory heat-stable chemical compound which is readily degraded by its own secreted enzyme.