• Title/Summary/Keyword: amino-polysaccharide

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From the Photosynthesis to Hormone Biosynthesis in Plants

  • Hyong Woo Choi
    • The Plant Pathology Journal
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    • v.40 no.2
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    • pp.99-105
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    • 2024
  • Land plants produce glucose (C6H12O2) through photosynthesis by utilizing carbon dioxide (CO2), water (H2O), and light energy. Glucose can be stored in various polysaccharide forms for later use (e.g., sucrose in fruit, amylose in plastids), used to create cellulose, the primary structural component of cell walls, and immediately metabolized to generate cellular energy, adenosine triphosphate, through a series of respiratory pathways including glycolysis, the tricarboxylic acid cycle, and oxidative phosphorylation. Additionally, plants must metabolize glucose into amino acids, nucleotides, and various plant hormones, which are crucial for regulating many aspects of plant physiology. This review will summarize the biosynthesis of different plant hormones, such as auxin, salicylic acid, gibberellins, cytokinins, ethylene, and abscisic acid, in relation to glucose metabolism.

Immunostimulating Activity and Characterization of Polysaccharides from Mycelium of Phellinus linteus

  • Lee, Jae Hoon;Soo Muk Cho;Kyung Sik Song;Sang Bae Han;Hwan Mook Kim;Nam Doo Hong;Ick Dong Yoo
    • Journal of Microbiology and Biotechnology
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    • v.6 no.3
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    • pp.213-218
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    • 1996
  • Hot-water extract, Fr. 1, of Phellinus linteus mycelium was fractionated into Fr. 2, 3, 4, and 5 by the difference of solubility in ethanol. The polysaccharide fractions were studied for their immunostimulating activity on in vitro T-independent polyc1onal antibody response to trinitrophenyl-haptened SRBC (sheep red blood cell). The Fr. 4 with the highest immunostimulating activity was subjected to DEAE-cellulose ion exchange chromatography and gave five fractions, 4-I, II, III, IV, and V. The in vitro immunostimulating assay of the five fractions showed that 4-I and 4-III had a similar activity to that of LPS but the other fractions had low activity. By analyses of chemical composition and HPLC, all fractions obtained were found to be heteropolysaccharide-protein complex. The molecular weights ranged from 9, 000 to 15, 000. Sugar analyses showed that glucose, galactose, mannose, arabinose, and xylose were main component. Uronic acid and amino sugar were also detected in the fractions. It should be noted that the molecular weight (15, 000) of 4-III was very small and the structure of 4-III may be different from the known immunostimulating branched $\beta$-(1longrightarrow3)-glucan.

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Toxic Components of Auricularia polytricha

  • Kim, Ha-Won
    • Archives of Pharmacal Research
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    • v.16 no.1
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    • pp.36-42
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    • 1993
  • To find biologically active components of the higher fungi of Korea, the carpophores of Auricularia polytricha, a well-known edible mushroom, were extracted with 0.14 M NaCl solution. The extract was successively fractionated by adding ammonium sulfate at various concentrations, and the respective precipitates were separated by centrifugation, then dialyzed and freeze-dried. When a does of 60 mg/kg of each was injected i.p. into ICR mice, the fraction which precipitated at 20% ammonium sulfate showed the highest toxicity, killing seven out of seven mice within two days. The fraction obtained at 40% ammonium sulfate showed the second highest toxicity. The two fractions were named auritoxin I and II after the genus name. However, they Nere shown to have nearly identical composition by physicochemical and 6.8% protein. The polysaccharide moiety was found to have 12.3% $\alpha$-linkage and 87.7% $\beta$-linkage and to be a heteromannoglucan consisting of 45.1% glucose, 435 mannose and 11.0% xylose. The protein moiety contained ten amino adids. The molecular weight of the toxin was $1.5\times10^6$ dalton by Sepharose CL-4B gel filtration. The modian lethal doses of auritoxin in mice were 56.4, 157.2 and 454.6 mg/kg by i.p., s.c. and p.o.administrations, respectively. The signs of intrxication were convulsion during the first 30 minutes after the injection, coma or sleeping within an hour, termor, lacrimation, nasal bleeding congestion, and death in 24 hours. Smong the various organs, the spleen was found to be enlarged remarkably. Human platelet aggregation was inhibited by the addition of auritoxin. The activity of malic dehydrogenase in vitro was inhibited by the toxin.

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Primary Structure of the Human VkII Regions Elicited by Haemophilus influenzae Type b Polysaccharide Vaccines; The J Gene Usage Is Restricted in Child Antibodies Using the A2 Gene

  • Yu, Kang-Yeol;Kim, Jin-Ho;Chung, Gook-Hyun
    • BMB Reports
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    • v.33 no.3
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    • pp.249-255
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    • 2000
  • The Haemophilus influenzae type b (Hib) has been a major cause of bacterial meningitis in children who are less than two years old. The variable (V) region repertoire of adult Caucasian antibodies (Abs) to Hib polysaccharide (PS) has been characterized well. The majority of adult antibodies against Hib uses VL that is derived from the Vk gene A2 and have arginine at the N region. In order to explore the possibility those antibody responses to Hib-PS is variable in various age groups, we examined the VL regions of the antibodies to Hib-PS in Korean adults and children. We immunized Korean adults (n = 8) and children (n = 39) with Hib tetanus conjugated vaccines, isolated RNAs from the peripheral lymphocytes, and amplified the A2-derived VL regions by RT-PCR. The PCR products were subcloned and sequenced. Forty-seven out of 54 independent clones from children used the $J{\kappa}2$, or $J{\kappa}3$ gene in preference. The adults, however, used all of the $J{\kappa}$ genes evenly. With respect to the amino acid sequences of variable regions, adult $A2-J{\kappa}$ recombinants have a germline sequence. But, the 76th codon (AGC) of child $A2-J{\kappa}2$ recombinants was substituted with CGC (arginine) in most cases (88 %) and 77 percent of child clones using the $A2-J{\kappa}3$ genes have isoleucine-109 at the junction of $J{\kappa}-C{\kappa}$ instead of threonine that is found in a germline sequence. These results suggest that the mechanism of antibody production in young children is different from that of adults.

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Studies on Constituents of the Higher Fungi of Korea (LV) -The Antitumor Components and Culture of Lyophyllum decastes- (한국산(韓國産) 고등(高等) 균류(菌類)의 성분(成分) 및 배양(培養)에 관한 연구(硏究)(제55보)(第55報) -Lyophyllum decastes의 항암(抗癌) 성분(成分) 및 배양(培養)-)

  • Lee, Chong-Ock;Kim, Hyung-Soo;Choi, Eung-Chil;Kim, Byong-Kak
    • Korean Journal of Pharmacognosy
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    • v.17 no.1
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    • pp.23-34
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    • 1986
  • To find antitumor components of Lyophyllum decastes, the mycelia of L. decastes were cultured in artificial media and a new antitumor component which showed potent antitumor activity against sarcoma 180 implanted in mice, was isolated and named 'Lyophyllan A'. Lyophyllan A was composed of a polysaccharide moiety (86%) and a protein moiety (2%). The polysaccharide moiety was found to be a heteroglycan which consisted of glucose (48.1%), mannose (30.8%), galactose, xylose and fucose. The protein moiety contained 18 amino acids. Cultural characteristics of L. decastes were investigated by shake culture method. The best result was obtained when L. decastes was cultured in medium glucose 50g, peptone 10g, corn steep liquor 30ml, $KH_2PO_4$ 0.87g, $MgSO_4$ $7H_2O$ 0.5g, $CaCl_2$ 0.3g, $FeSO_4{\cdot}7H_2O$ 10mg, $MnCl_2{\cdot}4H_2O$ 7mg, $ZnSO_4{\cdot}7H_2O$ 4mg and $CuSO_4{\cdot}5H_2O$ 1mg per one liter at $26^{circ}C$, 180rpm, for 9 days.

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Characterization of the rcsA Gene from Pantoea sp. Strain PPE7 and Its Influence on Extracellular Polysaccharide Production and Virulence on Pleurotus eryngii

  • Kim, Min Keun;Lee, Sun Mi;Seuk, Su Won;Ryu, Jae San;Kim, Hee Dae;Kwon, Jin Hyeuk;Choi, Yong Jo;Yun, Han Dae
    • The Plant Pathology Journal
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    • v.33 no.3
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    • pp.276-287
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    • 2017
  • RcsA is a positive activator of extracellular polysaccharide (EPS) synthesis in the Enterobacteriaceae. The rcsA gene of the soft rot pathogen Pantoea sp. strain PPE7 in Pleurotus eryngii was cloned by PCR amplification, and its role in EPS synthesis and virulence was investigated. The RcsA protein contains 3 highly conserved domains, and the C-terminal end of the open reading frame shared significant amino acid homology to the helix-turn-helix DNA binding motif of bacterial activator proteins. The inactivation of rcsA by insertional mutagenesis created mutants that had decreased production of EPS compared to the wild-type strain and abolished the virulence of Pantoea sp. strain PPE7 in P. eryngii. The Pantoea sp. strain PPE7 rcsA gene was shown to strongly affect the formation of the disease symptoms of a mushroom pathogen and to act as the virulence factor to cause soft rot disease in P. eryngii.

Modeling Studies of an Exotype Alginate Lyase Atu3025 from Agrobacterium Tumefaciens Strain C58, a Member of Polysaccharide Lyase Family 15

  • Kothandan, Gugan;Cho, Seung Joo
    • Journal of Integrative Natural Science
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    • v.3 no.2
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    • pp.72-77
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    • 2010
  • Alginate lyases, also known as alginases or alginate depolymerases, catalyze the degradation of alginate by a ${\beta}$-elimination mechanism that has yet to be fully elucidated. Alginate is a copolymer of ${\alpha}$-L-guluronate (G) and its C5 epimer ${\beta}$-D-mannuronate (M), arranged as homopolymeric G blocks, M blocks, alternating GM or random heteropolymeric G/M stretches. Almost all alginate lyases depolymerize alginate in an endolytical fashion via a ${\beta}$-elimination reaction. The alginate lyase Atu3025 from Agrobacterium tumefaciens strain C58, consisting of 776 amino-acid residues, is a novel exotype alginate lyase classified into polysaccharide lyase family 15. Till now there is no crystal structure available for this class of proteins. Since there is no template with high sequence identity, three-dimensional coordinates for exotype alginate lyase (PL 15 family) were determined using modeling methods (Comparitive modeling and Fold recognition). The structures were modeled using the X-ray coordinates from Heparinase protein family (PDB code: 3E7J). This enzyme (Atu3025) displays enzymatic activity for both poly-M and poly-G alginate. Since poly-M is widespread; docking of a tri-mannuronate against the modeled structure was performed. We identified some of those residues which are crucial for lyase activity. The results from this study should guide future mutagenesis studies and also provides a starting point for further proceedings.

Cloning, Expression, and Characterization of UDP-glucose Pyrophosphorylase from Sphingomonas chungbukensis DJ77

  • Yoon, Moon-Young;Lee, Kyoung-Jin;Park, Hea-Chul;Park, Sung-Ha;Kim, Sang-Gon;Kim, Sung-Kun;Choi, Jung-Do
    • Bulletin of the Korean Chemical Society
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    • v.30 no.6
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    • pp.1360-1364
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    • 2009
  • The bacterium Sphingomonas chungbukensis DJ77 produces the extracellular polysaccharide gellan in high yield. Gellan produced by this bacterium is widely used as a gelling agent, and the enzyme UDP-glucose pyrophosphorylase (UGP) is thought to play a key role in the gellan biosynthetic pathway. The UGP gene has been successfully cloned and over-expressed in E. coli. The expressed enzyme was purified with a molecular weight of approximately 32 kDa, as determined by a SDS-polyacrylamide gel, but the enzyme appears as ca. 63 kDa on a native gel, suggesting that the enzyme is present in a homodimer. Kinetic analysis of UDP-glucose for UGP indicates $K_m$ = 1.14 mM and $V_{max}$ = 10.09 mM/min/mg at pH 8.0, which was determined to be the optimal pH for UGP catalytic activity. Amino acid sequence alignment against other bacteria suggests that the UGP contains two conserved domains: An activator binding site and a glucose-1-phosphate binding site. Site-directed mutagenesis of Lys194, located within the glucose-1-phosphate binding site, indicates that substitution of the charge-reversible residue Asp for Lys194 dramatically impairs the UGP activity, supporting the hypothesis that Lys194 plays a critical role in the catalysis.

Generation of a Human Monoclonal Antibody to Cross-Reactive Material 197 (CRM197) and Development of a Sandwich ELISA for CRM197 Conjugate Vaccines

  • Kim, Dain;Yoon, Hyeseon;Kim, Sangkyu;Wi, Jimin;Chae, Heesu;Jo, Gyunghee;Yoon, Jun-Yeol;Kim, Heeyoun;Lee, Chankyu;Kim, Se-Ho;Hong, Hyo Jeong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.2113-2120
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    • 2018
  • Cross-reactive material 197 ($CRM_{197}$) is a non-toxic mutant of diphtheria toxin containing a single amino acid substitution of glycine 52 with glutamic acid. $CRM_{197}$ has been used as a carrier protein for poorly immunogenic polysaccharide antigens to improve immune responses. In this study, to develop a sandwich ELISA that can detect $CRM_{197}$ and $CRM_{197}$ conjugate vaccines, we generated a human anti-$CRM_{197}$ monoclonal antibody (mAb) 3F9 using a phage-displayed human synthetic Fab library and produced mouse anti-$CRM_{197}$ polyclonal antibody. The affinity ($K_D$) of 3F9 for $CRM_{197}$ was 3.55 nM, based on Bio-Layer interferometry, and it bound specifically to the B fragment of $CRM_{197}$. The sandwich ELISA was carried out using 3F9 as a capture antibody and the mouse polyclonal antibody as a detection antibody. The detection limit of the sandwich ELISA was <1 ng/ml $CRM_{197}$. In addition, the 3F9 antibody bound to the $CRM_{197}$-polysaccharide conjugates tested in a dose-dependent manner. This ELISA system will be useful for the quantification and characterization of $CRM_{197}$ and $CRM_{197}$ conjugate vaccines. To our knowledge, this study is the first to generate a human monoclonal antibody against $CRM_{197}$ and to develop a sandwich ELISA for $CRM_{197}$ conjugate vaccines.

Quality Characteristics of Rice Paper Roll with Frozen Rainbow Trout Oncorhynchus mykiss Meat Treated with Polysaccharide and TGase (다당류 및 TGase를 처리한 동결 무지개송어(Oncorhynchus mykiss)를 활용한 라이스페이퍼롤의 품질 특성)

  • Hye Min Park;Jong Bong Lee;Byoung Kyu An;Woo Jin Lee;Jung-Jin In;Hyeong Gu Han;Seung Ah Son;Yeon Joo Bae;Kil Bo Shim
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.56 no.5
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    • pp.585-595
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    • 2023
  • This study was conducted to optimize the rice paper roll processing conditions with frozen rainbow trout Oncorhynchus mykiss meat (RPR-FRT) treated with polysaccharide and TGase using a response surface methodology and to examine their quality characteristics. The RSM results for the RPR-FRT showed that the optimum condition for the garlic-pepper mixture was 21.2 g and that for starch was 22.6 g based on 150 g of RPR-FRT. The RPR-FRT contained 58.47 g/100 g of moisture, 8.57 g/100 g of crude protein, 3.28 g/100 g of crude lipid, and 1.00 g/100 g of ash. The vitamin E content was 1,010.91 ㎍/100 g. Based on their contents, the samples could be considered good supplements for P, Cr, and Se. The RPR-FRT contained unsaturated fatty acids (75.84%), DHA (10.33%), and EPA (2.56%). Anserine, arginine, glycine, and taurine accounted for 41.93%, 11.27%, 7.13%, and 7.00% of free amino acids in the RPR-FRT, respectively. The sulfur compounds in the RPR-FRT constituted 73.16% of the total flavor compounds. The RPR-FRT prepared using the optimum conditions was superior in masking off-flavor and showed improved nutritional content.