• Title/Summary/Keyword: Glycosyl-linkage

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Chemical Characteristics and Immuno-stimulating Properties of Biopolymers Extracted from Acanthopanax sessiliflorus

  • Jeong, Sang-Chul;Jeong, Young-Tae;Yang, Byung-Keun;Song, Chi-Hyun
    • BMB Reports
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    • v.39 no.1
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    • pp.84-90
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    • 2006
  • During our search for macrophage stimulating compounds from medicinal plants, we isolated biopolymers from Acanthopanax sessiliflorus. Isolated fraction AS-5 showed maximum potential, and stimulated lysosonal enzymatic activity by 230% at $300\;{\mu}g/ml$. The nitric oxide (NO) producing ability of AS-5 $100\;{\mu}g/ml$ was $58\;{\mu}M$ when treated with interferon-$\gamma$ and lipopolysaccharide $20\;{\mu}g/ml$. The lymphocyte proliferating effects of isolated biopolymer fractions were also investigated. Highest lymphoproliferative activity (a 2.8-fold enhancement compared to saline treated group was exhibited by AS-3 at $200\;{\mu}g/ml$ followed by AS-5 and AS-6. The AS-3 fraction stimulated only T-lymphocytes and had little or no effect on B-lymphocyte proliferation. Partially methylated alditol acetates were prepared to elucidate the glycosyl linkage-compositions of the AS-3 and AS-5 biopolymers, and were analyzed by GC-MS. The AS-3 and AS-5 biopolymer fractions were found to contain 2,3,4-tri-O-methyl-D-glucitol, 2,3,4-tri-O-methyl-D-galacitol 3,4,6-tri-O-methyl-galacitol, 2-O-methyl-arabinitol and 2,4,6-tri-O-methyl-D-glucitol, 2,3,6-tri-O-methyl-D-galacitol linkages, respectively.

Glycosyl-linkages of Acid Soluble Polysaccharide from Green Laver, Enteromopha prolifera (가시파래 산성 수용성 다당의 구성당 결합 특성)

  • Koo Jae Geun;Choi Yong Seok;Ha Jin Hwan
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.5
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    • pp.524-528
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    • 2002
  • Green layer, Enteromorpha prolifera, is regarded as one of important materials for food processing in Korea. The acidic water-soluble polysaccharide (CPC-PS) isolated from the alga with hot water and cetylpyridium chloride was mainly constituted of rhamnose, xylose, uronic acid and sulfate. To determine the glycosyl-linkages and positions of sulfate by methylation, the CPC-PS was reduced and/or sulfates. A marked increase of glucose content in the reduced polysaccharide indicated that glucuronic acid was a major sugar in the polymer and sulfation was deduced to occur on O-3 of rhamnose and O-2 of xylose. According to the methylation analysis of the native, reduced, desulfated and reduced-desulfated polymers, CPC-PS mainly composed of 1,4- and 1,2,3-linked rhamnose 3-sulfate, 1,4-linked xylose 2-sulfate, 1,4-linked xylose and 1,4-linked glucuronic acid. Minor 1,4-linked rhamnose and 1,4,6-linked galactose residues were also detected.

Identification and molecular characterization of the chitinase gene, EaChi, from the midgut of the earthworm, Eisenia andrei (붉은줄지렁이 (Eisenia andrei) 중장에서 발현되는 chitinase 유전자, EaChi의 동정 및 분자생물학적 특성에 관한 연구)

  • Tak, Eun Sik;Kim, Dae hwan;Lee, Myung Sik;Ahn, Chi Hyun;Park, Soon Cheol
    • Journal of the Korea Organic Resources Recycling Association
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    • v.18 no.3
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    • pp.31-37
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    • 2010
  • Chitinases (EC 3.2.1.14) hydrolyze the ${\beta}$-1,4-linkages in chitin, the second most abundant polymer of N-acetyl-${\beta}$-D-glucosamine which is a structural component of protective biological matrices such as fungal cell walls and insect exoskeletons. The glycosyl hydrolases 18 family including chitinases is an ancient gene family widely expressed in archea, prokaryotes and eukaryotes. Since earthworms live in the soil with a lot of microbial activities and fungi are supposed to be a major component of the diet of earthworm, it has been reported that there would be appropriate immune system to protect themselves from microorganisms attacks. In this study, the novel chitinase, EaChi, from the midgut of earthworm, Eisenia andrei, were identified and characterized. To obtain full-length cDNA sequence of chitinase, RT-PCR and RACE-PCR analyses were carried out by using the previously identified EST sequence amongst cDNA library established from the midgut of E. andrei. EaChi, a partial chitinase gene, was composed of 927 nucleotides encoding 309 amino acids. By the multiple sequence alignments of amino acids with other different species, it was revealed that EaCHI is a member of glycosyl hydrolases 18 family, which has two highly conserved domains, substrate binding and catalytic domain.

The effect of Far-infrared on survival rate and genetic mutation of Escherichia-coli (원적외선이 대장균의 생존율과 돌연변이에 미치는 영향)

  • Kim, Jae-Yoon;Park, Rae-Joon
    • The Journal of Korean Physical Therapy
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    • v.13 no.3
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    • pp.637-642
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    • 2001
  • Hershel found out Infrared for the first time, in the Industrial Revolution the Infrared and FlR had been begun to use making products. FIR with low temperature can deeply penetrate on the human body composed things without troublesome. since FIR has effectively operated on the human body at low temperature (35-40$^{\circ}$C). When FIR penetrated on the human body, it would inhibit the abnormal genes and cells expression, and then information of DNA and RNA would be reexpressed for arranging DNA and RNA abnormal state. As FIR's receptors in the body, it colud be presumed that N-glycosyl linkage of purine and deoxyribose, RNA splicing process. and Heat shock protein. To radiate optimized FIR, in this study, we made the FIR radiation compound and instrument for unharming biological things. According to the results, the FIR radiation to E. coli., it did not induce genetic mutations and change the survival rate of E.coli.

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Recent Studies on the Chemical Constituents of Korean Ginseng (Panax ginseng C. A. Meyer) (고려인삼의 화학성분에 관한 고찰)

  • 박종대
    • Journal of Ginseng Research
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    • v.20 no.4
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    • pp.389-415
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    • 1996
  • Panax ginseng C.A. Meyer(Araliaceae) has been traditionally used as an expensive and precious medicine in oriental countries for more than 5, 000 years. Ginseng saponin isolated from the root of Panax ginseng have been regarded as the main effective components responsible for the pharmacological and biological activities. Such as antiaging effects. antidiabetic effects anticancer effects. Protection against physical and chemical stress. Analgesic and antipyretic effects. Effects on the central nervous system, tranquilizing action and others. Thirty kinds of ginsenosides have been so far isolated from ginseng saponin and their chemical structures have been elucidated since 1960's. Among which protopanaxadiol type is 19 kinds. protopanaxatriol type. 10 kinds and oleanane type, one. Since ginsenosides are generally labile under acidic conditions ordinary acid hydrolysis is always accompanied by many side reactions, such as epimerization. hydroxylation and cyclization of side chain of the sapogenins Especially. it is well known that C-20 glycosyl linkage of ginsenoside was hydrolysed on heating with acetic acid to give an equilibrated mixture of 20(S) and 20(R) epimers. And also, the chemical transformations of the secondary metabolites have appeared during the steaming process to prepare red ginseng. Indicating demalonylation of malonyl ginsenosides, elimination of glycosyl residue at C-20 and isomerization of hydroxyl configuration at C-20. But these studies have not provided a comprehensive picture in explaning how these ginsenosides showed val'iotas pharmacological activities of ginseng. Though some of them have been involved in the mechanism of pharmacological actions. Recently, non-saponin components have received a great deal of attention for their antioxidant, anticancer antidiabetic, immunomodulating. anticomplementary activities and so on. To meet the demand for such wide applications, studies on the non-saponin components play an important role in providing a good evidence of pharmacological and biol ogical activities. Among the non-saponin constituents of Korean ginseng, polyacetylenes, phenols. Sesquiterpenes, alkaloids. polysaccharides oligosaccharides, oligopeptides and aminoglycosides together with ginsenosides of terrestrial part are mainly described.

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Mechanism of Far-infrared how to affect the human body (원적외선의 인체작용메카니즘)

  • Kim, Jae-Yoon;Park, Young-Han;Park, Don-Mork;Park, Rae-Joon
    • The Journal of Korean Physical Therapy
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    • v.13 no.2
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    • pp.477-482
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    • 2001
  • Until now, it has not been well known for Far-infrared(FIR) how to affect to the human body. We introduced and presumed the mechanism of FIR based on molecular biology in this study, as below. The human body is composed of proteins which get easily changed by a thermal factor (about 42 $^{\circ}$C over). FIR with low temperature can deeply penetrate on the human body composed things without troublesome, since FIR has effectively operated on the human body at low temperature (35-40 $^{\circ}$C). When FIR penetrated on the human body, it would inhibit the abnormal genes and cells expression, and then information of DNA and RNA would be reexpressed for arranging DNA and RNA abnormal state. As FIR's receptors in the body, it colud be presumed that N-glycosyl linkage of purine and deoxyribose, RNA splicing process, and heat shock protein.

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Chitinase-producing Salinivibrio bacteria isolated from salt-fermented shrimp with antimicrobial and safety assessments

  • Le, Bao;Chung, Gyuhwa;Yang, Seung Hwan
    • Journal of Applied Biological Chemistry
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    • v.61 no.3
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    • pp.233-238
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    • 2018
  • Chitinases are glycosyl hydrolases which cleave the ${\beta}$-1,4 linkage of chitin into oligo or monomers of N-acetylglucosamine. These bacterial enzymes have been used for a wide range of applications in the food and pharmaceutical industries. In this study, we isolated two potential chitinolytic strains, BAO-01 and BAO-02, from salt-fermented shrimp, which were shown to belong to the genus Salinivibrio through genetic characterization using 16S rRNA. These isolates were gram-positive, rod-shaped, and non-spore forming. BAO-01 showed greater growth and chitinase activity than BAO-02 after the incubation at $37^{\circ}C$ for 4 days. Both strains grew on a wide range of carbon and nitrogen sources, pH values, temperatures, and salt levels. However, they showed minor biochemical differences. In addition, their antimicrobial activities against foodborne pathogens and antibiotic susceptibilities were evaluated. These Salinivibrio spp. did not show bioamine production, hemolytic activity, and mucin degradation. Therefore, the in vitro screening results suggested that these bacteria could be widely used as new candidates for chitin hydrolyzation and seafood fermentation.

Mutational Analysis of Thermus caldophilus GK24 ${\beta}$-Glycosidase: Role of His119 in Substrate Binding and Enzyme Activity

  • Oh, Eun-Joo;Lee, Yoon-Jin;Choi, Jeong-Jin;Seo, Moo-Seok;Lee, Mi-Sun;Kim, Gun-A;Kwon, Suk-Tae
    • Journal of Microbiology and Biotechnology
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    • v.18 no.2
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    • pp.287-294
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    • 2008
  • Three amino acid residues (His119, Glu164, and Glu338) in the active site of Thermus caldophilus GK24 ${\beta}$-glycosidase (Tca ${\beta}$-glycosidase), a family 1 glycosyl hydrolase, were mutated by site-directed mutagenesis. To verify the key catalytic residues, Glu164 and Glu338 were changed to Gly and Gln, respectively. The E164G mutation resulted in drastic reductions of both ${\beta}$-galactosidase and ${\beta}$-glucosidase activities, and the E338Q mutation caused complete loss of activity, confirming that the two residues are essential for the reaction process of glycosidic linkage hydrolysis. To investigate the role of His119 in substrate binding and enzyme activity, the residue was substituted with Gly. The H119G mutant showed 53-fold reduced activity on 5mM p-nitrophenyl ${\beta}$-D-galactopyranoside, when compared with the wild type; however, both the wild-type and mutant enzymes showed similar activity on 5mM p-nitrophenyl ${\beta}$-D-glucopyranoside at $75^{\circ}C$. Kinetic analysis with p-nitrophenyl ${\beta}$-D-galactopyranoside revealed that the $k_{cat}$ value of the H119G mutant was 76.3-fold lower than that of the wild type, but the $K_m$ of the mutant was 15.3-fold higher than that of the wild type owing to the much lower affinity of the mutant. Thus, the catalytic efficiency $(k_{cat}/K_m)$ of the mutant decreased to 0.08% to that of the wild type. The $k_{cat}$ value of the H119G mutant for p-nitrophenyl ${\beta}$-D-glucopyranoside was 5.l-fold higher than that of the wild type, but the catalytic efficiency of the mutant was 2.5% of that of the wild type. The H119G mutation gave rise to changes in optima pH (from 5.5-6.5 to 5.5) and temperature (from $90^{\circ}C\;to\;80-85^{\circ}C$). This difference of temperature optima originated in the decrease of H119G's thermostability. These results indicate that His119 is a crucial residue in ${\beta}$-galactosidase and ${\beta}$-glucosidase activities and also influences the enzyme's substrate binding affinity and thermostability.

Research of human body information interfacing with Far infrared and application to physical therapy (Far infrared를 이용한 생체정보 인터페이싱에 대한 연구)

  • Park, Rae Joon;Kim, Jae-Yoon
    • The Journal of Korean Physical Therapy
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    • v.13 no.3
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    • pp.509-527
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    • 2001
  • The Sun's ray is composed of Infrared(49%), Visible light(40%) and Ultra violet(11%), however the ray getting to the earth is FIR(Far infrared; 60%), IR(Infrared; 20%), and UV(Ultra Violet; 20%). Human beings has utilized FIR already from time immemorial. Hershel found out Infrared for the first time. in the Industrial Revolution the Infrared and FIR had been begun to use making products. In these days, with contemporary science FIR would be begun to clear up the implication in the human body and organic compound. IR classified by wavelength three parts NlR, MIR, FIR. There is FIR which is radiated from healthy human body the wave length is 8-l4m. The human body is composed of proteins which get easily changed by a thermal factor (about 42 $^{\circ}$C over). FIR with low temperature can deeply penetrate on the human body composed things without troublesomes, since FIR has effectively operated on the human body at low temperature (35-40 $^{\circ}$C). When FlR penetrated on the human body. it would inhibit the abnormal genes and cells expression, and then information of DNA and RNA would be reexpressed for arranging DNA and RNA abnormal state. As FlR's receptors in the body, it could be presumed that N-glycosyl linkage of purine and deoxyribose, RNA splicing process, and Heat shock protein. To take the FIR which was a optimized wavewlength and strength, at first, we induced the characteristic algorithm and the computerized programing. Then we formed that the formular of optimized FIR with physical, mathematical logic and theory. especially, Plank, Kirchhoff, Wien, Stefan-Boltzmann's logic and law. In the long run, the formular was induced with integration mathematical, since we had to know the molecular wavelength. Based on the induced formular as above, we programmed the optimized FlR radiating computerized program. In this research, we designed the eletronic circuit f3r interfacing with human body to diagnosis and treatment with FIR sensor which radiated FIR wavelength optimized.

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Cloning and Expression of a Thermostable ${\alpha}$-Galactosidase from the Thermophilic Fungus Talaromyces emersonii in the Methylotrophic Yeast Pichia pastoris

  • Simila, Janika;Gernig, Anita;Murray, Patrick;Fernandes, Sara;Tuohy, Maria G.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1653-1663
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    • 2010
  • The first gene (${\alpha}$-gal1) encoding an extracellular ${\alpha}$-Dgalactosidase from the thermophilic fungus Talaromyces emersonii was cloned and characterized. The ${\alpha}$-gal1 gene consisted of an open reading frame of 1,792 base pairs interrupted by six introns that encoded a mature protein of 452 amino acids, including a 24 amino acid secretory signal sequence. The translated protein had highest identity with other fungal ${\alpha}$-galactosidases belonging to glycosyl hydrolase family 27. The ${\alpha}$-gal1 gene was overexpressed as a secretory protein with an N-terminal histidine tag in the methylotrophic yeast Pichia pastoris. Recombinant ${\alpha}$-Gal1 was secreted into the culture medium as a monomeric glycoprotein with a maximal yield of 10.75 mg/l and purified to homogeneity using Hisbinding nickel-agarose affinity chromatography. The purified enzyme was maximally active at $70^{\circ}C$, pH 4.5, and lost no activity over 10 days at $50^{\circ}C$. ${\alpha}$-Gal1 followed Michaelis-Menten kinetics ($V_{max}\;of\;240.3{\mu}M/min/mg,\;K_m\;of\;0.294 mM$) and was inhibited competitively by galactose ($K_m{^{obs}}$ of 0.57 mM, $K_i$ of 2.77 mM). The recombinant T. emersonii ${\alpha}$-galactosidase displayed broad substrate preference, being active on both oligo- and polymeric substrates, yet had strict specificity for the ${\alpha}$-galactosidic linkage. Owing to its substrate preference and noteworthy stability, ${\alpha}$-Gal1 is of particular interest for possible biotechnological applications involving the processing of plant materials.