• Title/Summary/Keyword: N-Acetyl-Glucosamine

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Action Patterns of Chitinase and Separations of Chitooligosaccharides Produced by Chitinolytic Hydrolysis (키티나제에 의한 키토올리고당의 생성활성 규명과 올리고당의 당별 분리 생산)

  • Kim, Kwang
    • KSBB Journal
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    • v.17 no.1
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    • pp.100-105
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    • 2002
  • N-acetyl-D-glucosamine oligosaccharides [(GlcNAc)n] whose degree of polymer-ization is from one to ten (n=1-10) were fractionated by column chromatography on CM-Sephadex. Electro dialysis from a partially deacetylated chitosan hydrolysate prepared crudely with the N-acetyl-D-glucosaminidase(chitinase) and exo-N, N'-diacetylchito-biohydrolase(chitobiase) of Serratia marcescens QM B1466. Reducing sugar compositions and sequences of the N-acetyl-glucosamine oligosaccharides were identified by N-acetylation, randomly cleavage with chitinase and ego-splitting with chitobiase. N-acetyl-glucosamine heterochitooligosaccharides with glucosamine oligosaccharides, (GlcN)n at the reducing end residues together with $(GlcN)_1\sim(GlcN)_4$ were detected. Separation was accomplished by prefractionation with election by 0 to 1.0 M NaCl gradient solution. $(GlcNAc)_1 =4.25%,\; (GlcNAc)_2=4.49%,; (GlcNAc)_3=11.1%,\; (GlcNAc)_4=2.5%,$$ $(GlcNAc)_{5}$=0.64%, $(GlcNAc)_{6}$=2.12% and $(GlcNAc)_{7}$=1.21%, respectively, were crystallized after electrodialysis and lyophilization Each N-acetyl-D-glucosamine oligosaccharides content were detected by HPLC.

Effects of Poly-N-acetyl Glucosamine(pGlcNAc) Patch on Wound Healing in db/db Mouse (Poly-N-acetyl-glucosamine이 당뇨병 쥐에서 창상치료에 미치는 영향)

  • Yang, Ho Jik;Yoon, Chi Sun
    • Archives of Plastic Surgery
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    • v.35 no.2
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    • pp.121-126
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    • 2008
  • Purpose: Poly-N-acetyl glucosamine(PGlcNAc) nanofiber-based materials, produced by a marine microalga, have been characterized as effective hemostatic and angiogenic agents. The similarity between PGlcNAc patch and the natural extracellular matrix allows it to support new healthy tissue growth in an injured area and to encourage fluid absorption. In this study, we hypothesized that a poly-N-acetyl glucosamine fiber patch(PGlcNAc patch) may enhance wound healing in the db/db mouse. Methods: PGlcNAc patches were applied on one square centimeter, full-thickness, skin wounds in the db/db mouse model. Wounds(n=15 per group) were dressed with a PGlcNAc nanofiber patch for 1 hour(1 h), 24 hours(24 h) or left untreated(NT). After the application time, patches were removed and wounds were allowed to heal spontaneously. The rate of wound closure was evaluated by digital analysis of unclosed wound area in course of time. At day 10, wounds(n=7 per group) were harvested and quantified with immunohistochemical markers of proliferation(Ki-67) and vascularization (platelet endothelial cell adhesion molecule, PECAM-1). Results: Wounds dressed with PGlcNAc patches for 1 hour closed faster than control wounds, reaching 90% closure in 16.6 days, nine days faster than untreated wounds. Granulation tissue showed higher levels of proliferation and vascularization following 1 h treatment than the 24 h and NT groups. In addition to its hemostatic properties, the PGlcNAc material also appears to accelerate wound closure in healing-impaired genetically diabetic mice. Conclusion: This material, with its combination of hemostatic and wound healing properties, has the potential to be effective agent for the treatment of complicated wounds.

Characterization and Distribution of Glycoconjugates in the Intestines of Sebastes schlegeli, Halichoeres poecilopterus, Bryzoichthys lysimus, and Takifugu pardalis. (조피볼락, 용치놀래기, 송곳니베도라치 및 졸복 장관의 복합당질 성상 및 분포)

  • Jeong, Gil-Nam
    • Journal of Life Science
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    • v.18 no.6
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    • pp.782-788
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    • 2008
  • The aim of this work is to show differences in the pattern of glycoconjugate composition in the intestines of four teleostean species (Sebastes schlegeli, Halichoeres poecilopterus, Bryzoichthys lysimus, and Takifugu pardalis). We compared four regions of all species studied. The specimens were processed and stained with nine kinds of biotinylated lectins (DBA, SBA, PNA, BSL- I , RCA- I , sWGA, UEA- I , LCA and Con A). Except for Sebastes schlegeli, no differences between regions were observed. The intestinal epithelium of Halichoeres poecilopterus possessed D-glucose/mannose residues in all regions. ${\beta}$-N-acetyl-D-galactosamine was distinctive along the intestines, although the pattern of diversity was different in Sebastes schlegeli, Bryzoichthys lysimus, and Takifugu pardalis. Additionally, the occurrence of Galactose-${\beta}$-1,3-N-acetyl-D-galactosamine and ${\alpha}$-D-galactose were confirmed in the proximal, middle, and distal intestine of Sebastes schlegeli, while rectal intestine lacked these sugar residues. Along with ${\beta}$-N-acetyl-D-galactosamine, ${\beta}$-N-acetyl-D-glucosamine and D-glucose/mannose were also determined in Bryzoichthys lysimus. Galactose-${\beta}$-1,3-N-acetyl-D-galactosamine, D-galactose, and D-glucose/mannose were also present in Takifugu pardalis.

Regional Difference in Distribution of Glycoconjugates in Mouse Epididymis (생쥐 부정소 부위별 당쇄 분포의 차이)

  • 계명찬
    • Development and Reproduction
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    • v.5 no.2
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    • pp.167-173
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    • 2001
  • To characterize the difference in glycoconjugates of mouse epididymis, lectin labeling of the tissue section was conducted using Ulex europaeus agglutinin I(UEA I), succinylated wheat germ agglutinin(sWGA), and Griffonia simplicifolia lectin-I(GSL-I). UEA I which binds to outer $\alpha$-L-fucose residue that is a terminal sugar of the side chain branched from oligosaccharide chain gave the labeling in the proximal caput epithelia exclusively. Lumen was commonly labeled in all of the organ. It suggested that the glycoconjugates bearing outer $\alpha$ -L-fucose residue were largely expressed in the initial segments ot epididymis and subjected to secretion. GSL-I which binds to terminal $\alpha$ -D-galactosyl residue of glycoconjugates gave the labeling in the cytoplasm of clear cells and basal cells, and cilia in corpus and cauda regions but not in the caput region. There was no vast difference in labeling pattern by sWGA which binds to N-acetyl-glucosamine residue among the epididymal regions. Clear cells in corpus and cauda epithelia showed more intense labeling by sWGA compared to principal cells, suggesting the functional specialization of this type of cells. The labeling intensities of luminal content by UEA I and sWGA decreased in cauda region compared to corpus region suggesting the presence of enzymatic activities responsible for processing the $\alpha$-L-fucose and N-acetyl-glucosamine residues from secreted glycoconjugates. In summary, the difference in glycoconjugates bearing the $\alpha$-L-fucose, $\alpha$-D-galactose, and N-acetyl-glucosamine residues according to the type of epithelial cells and epididymal segments suggests functional specialization and different roles of each segment in the processing of sperm surface antigens during the epididymal transit.

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Glucosamine Hydrochloride and N-Acetylglucosamine Influence the Response of Bovine Chondrocytes to TGF-β3 and IGF in Monolayer and Three-Dimensional Tissue Culture

  • Pizzolatti, Andre Luiz A.;Gaudig, Florian;Seitz, Daniel;Roesler, Carlos R.M.;Salmoria, Gean Vitor
    • Tissue Engineering and Regenerative Medicine
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    • v.15 no.6
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    • pp.781-791
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    • 2018
  • BACKGROUND: Glucosamine hydrochloride (GlcN HCl) has been shown to inhibit cell growth and matrix synthesis, but not with N-acetyl-glucosamine (GlcNAc) supplementation. This effect might be related to an inhibition of critical growth factors (GF), or to a different metabolization of the two glucosamine derivatives. The aim of the present study was to evaluate the synergy between GlcN HCl, GlcNAc, and GF on proliferation and cartilage matrix synthesis. METHOD: Bovine chondrocytes were cultivated in monolayers for 48 h and in three-dimensional (3D) chitosan scaffolds for 30 days in perfusion bioreactors. Serum-free (SF) medium was supplemented with either growth factors (GF) $TGF-{\beta}$ ($5ng\;mL^{-1}$) and IGF-I ($10ng\;mL^{-1}$), GlcN HCl or GlcNAc at 1mM each or both. Six groups were compared according to medium supplementation: (a) SF control; (b) SF + GlcN HCl; (c) SF + GlcNAc; (d) SF + GF; (e) SF + GF + GlcN HCl; and (f) SF + GF + GlcNAc. Cell proliferation, proteoglycan, collagen I (COL1), and collagen II (COL2) synthesis were evaluated. RESULTS: The two glucosamines showed opposite effects in monolayer culture: GlcN HCl significantly reduced proliferation and GlcNAc significantly augmented cellular metabolism. In the 30 days 3D culture, the GlcN HCl added to GF stimulated cell proliferation more than when compared to GF only, but the proteoglycan synthesis was smaller than GF. However, GlcNAc added to GF improved the cell proliferation and proteoglycan synthesis more than when compared to GF and GF/GlcN HCl. The synthesis of COL1 and COL2 was observed in all groups containing GF. CONCLUSION: GlcN HCl and GlcNAc increased cell growth and stimulated COL2 synthesis in long-time 3D culture. However, only GlcNAc added to GF improved proteoglycan synthesis.

Anti-wrinkle Effect of N-Acetyl-D-glucosamine (NAG) (N-Acetyl-D-glucosamine (NAG)의 피부주름 개선 효과)

  • Kim Kwang Soo;Choi Gun Ho;Choi Jang Woo;Choi Jun Hak;Han Song Hee;Nam Sang Yun;Lee Seung Hwa
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.4 s.48
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    • pp.457-462
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    • 2004
  • Anti-wrinkle effect of N-Acetyl-D-glucosamine (NAG) was evaluated by collagen synthesis and proliferation of normal human fibroblast. NAG was obtained by purifying deacetylated chitin which can be derived from chitin-rich crab shell. We studied in in-vitro cultures of human normal fibroblast, whether synthesis of collagens and fibroblast growth activation in these cells can be enhanced in the presence of NAG. It 야d not show any adverse effects in human shin irritation patch test. In in-vivo mouse test, it showed anti-wrinkle effect in hairless mouse (6W/F). From the HPLC analysis, the stability of NAG in the cosmetics product could be maintained for a long time. These results demonstrated that NAS can be useful anti-wrinkle cosmetic ingredient.

Cloning and Phylogenetic Analysis of Chitin Synthase Genes from Tricholoma matsutake

  • Suh, Seok-Jong;Kim, Il-Hyeon;Nam, Ju-Hyun;Ghim, Sa-Youl;Bae, Kyung-Sook;Kim, Jong-Guk
    • Mycobiology
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    • v.29 no.4
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    • pp.179-182
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    • 2001
  • Chitin synthases(UDP-N-acetyl-D-glucosamine: chitin 4-$\beta$-N-acetyl-D-glucosaminyl transferase, EC 2.4.1.16) catalyze the synthesis of chitin from UDP-N-acetyl-D-glucosamine. Two zymogenic type of chitin synthase gene(TmCHS1 and TmCHS2) were amplified and its nucleotide sequences were determined. By the amino acid comparison and UPGMA tree grouping, TmChs1 and TmChs2 were classified as class II and class IV chitin synthases respectively. The class II type TmChs1 was grouped with others of Agaricales ectomycorrhizal mushroom. Additionally the phylogenetic tree was well adapted to Hymenomycete previously classified by morphological and physiological characteristics.

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Effective Production of N-Acetyl-$\beta$-glucosamine by Serratia marcescens Using Chitinadceous Waste

  • Kim, Kwang;A. Louise Creagh;Charles A. Haynes
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.2
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    • pp.71-77
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    • 1998
  • The strain of Serratia marcescens QM B1466 produces selectively large amount of chitinolytic enzymes (about 1mg/L medium). Enzymatic hydrolysis of chitin to N-acetyl-${\beta}$-D-glucosamine (NAG) was performed with a system consisting of two hydrolases (chitinase and chitobiase) produced by optimization of a microbial host consuming chitin particles. For the development of Large-scale biological process for the production of NAG from chitinaceous waste, the selection and optimization of a microbial host, particle size of crab/shrimp chitin sources and initial induction time using chitin as a sole carbon source on chitinase/chitobiase production and NAG production were examined. Crab-shell chitin(1.5%) treated by dilute acid and , ball-milled with a normal diameter less than 250m gave the highest chitinase activity over a 7 days culture. Crude chitinase/ chitobiase solution obtained in a 10 L fed-batch fermentation showed a maximum activities of 23.6 U/mL and 5.1 U/mL, respectively with a feeding time of 3 hrs, near pH 8.5 at 30$^{\circ}C$.

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Studies on Glycolipids in Bacteria -Part II. On the Structure of Glycolipid of Selenomonas ruminantium- (세균(細菌)의 당지질(糖脂質)에 관(關)한 연구(硏究) -제2보(第二報) Selenomonas ruminantium의 당지질(糖脂質)의 구조(構造)-)

  • Kim, Kyo-Chang
    • Applied Biological Chemistry
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    • v.17 no.2
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    • pp.125-137
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    • 1974
  • The chemical structure of glycolipid of Selenomonas ruminantium cell wall was to be elucidated. The bacterial cells were treated in hot TCA and the glycolipid fractions were extracted by the solvent $CHCl_3\;:\;CH_3OH$ (1 : 3). The extracted glycolipids fraction was further separated by acetone extraction. The acetone soluble fraction was named as the spot A-compound. The acetone insoluble but ether soluble fraction was named as the spot B-compound. These two compounds were examined for elucidation of their chemical structure. The results were as follows: 1. The IR spectral analysis showed that O-acyl and N-acyl fatty acids were linked to glucosamine moiety in the spot A-compound. However in the spot B-compound in addition to O and N-acyl acids phosphorus was shown to be attached to glucosamine. 2. It was recognized by gas liquid chromatography that spot A compound contained beta-OH $C_{13:0}$ fatty acid in predominance in addition to the fatty acid with beta-OH $C_{9:0}$, whereas the spot B compound was composed of the predominant fatty acid of beta-OH $C_{13:0}$ with small amount of beta-OH $C_{9:0}$. 3. According to the paper chromatographic analysis of hydrazinolysis products of the spot A compound, a compound of a similar Rf value as the chitobiose was recognized, which indicated a structure of two molecules glucosamine condensed. The low Rf value of the hydrazinolysis product of the spot B-compound confirmed the presence of phosphorus attached to glucosamine. 4. The appearance of arabinose resulting from. ninhydrin decomposition of the acid hydrolyzate of the spot A compound indicated that the amino group is attached to $C_2$ of glucosamine. 5. The amount of glucosamine in the N-acetylated spot A compound decreased in half of the original content by the treatment. with $NaBH_4$, indicating that there are two molecules of glucosamines in the spot A compound. The presence of 1, 6-linkage between two molecules of glucosamine was suggested by the Morgan-Elson reaction and confirmed by the periodate decomposition test. 6. By the action of ${\beta}-N-acetyl$ glucosaminidase the N-acetylated spot A compound was completely decomposed into N-acetyl glucosamine, whereas the spot B compound was not. This indicated the spot A compound has a beta-linkage. 7. When phosphodiesterase or phosphomonoesterase acted on $^{32}P-labeled$ spot B compound, $^{32}P$ was not released by phosphodiesterase, but completely released by phosphomonoesterase. This indicated that one phosphorus is linked to glucosamine moiety. 8. The spot A compound is assumed to have the following chemical structure: That is glucosaminyl, ${\beta}-1$, 6-glucosamine to which O-acyl and N-acyl fatty acids are linked, of which the predominant fatty acid is beta-OH $C_{13:0}$ fatty acid in addition to beta-OH $C_{9:0}$ fatty acid 9. The spot B compound is likely to have the linkage of $glucosaminyl-{\beta}-1$, 6-glucosamine to which phosphorus is linked in monoester linkage. Furthermore both O-acyl and N-acyl fatty acids contained beta-OH $C_{13:0}$ fatty acid predominantly in addition to beta-OH $C_{9:0}$ fatty acid.

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