• Title/Summary/Keyword: sugar chain

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Role of Osmotic and Salt Stress in the Expression of Erythrose Reductase in Candida magnoliae

  • Park, Eun-Hee;Lee, Ha-Yeon;Ryu, Yeon-Woo;Seo, Jin-Ho;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • v.21 no.10
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    • pp.1064-1068
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    • 2011
  • The osmotolerant yeast, Candida magnoliae, which was isolated from honeycomb, produces erythritol from sugars such as fructose, glucose, and sucrose. Erythrose reductase in C. magnoliae (CmER) reduces erythrose to erythritol with concomitant oxidation of NAD(P)H. Sequence analysis of the 5'-flanking region of the CmER gene indicated that one putative stress response element (STRE, 5'-AGGGG-3'), found in Saccharomyces cerevisiae, exists 72 nucleotides upstream of the translation initiation codon. An enzyme activity assay and semiquantitative reverse transcription polymerase chain reaction revealed that the expression of CmER is upregulated under osmotic and salt stress conditions caused by a high concentration of sugar, KCl, and NaCl. However, CmER was not affected by osmotic and oxidative stress induced by sorbitol and $H_2O_2$, respectively. The basal transcript level of CmER in the presence of sucrose was higher than that in cells treated with fructose and glucose, indicating that the response of CmER to sugar stress is different from that of GRE3 in S. cerevisiae, which expresses aldose reductase in a sugarindependent manner. It was concluded that regulation of CmER differs from that of other aldose reductases in S. cerevisiae.

Isolation and Characterization of Intestinal Immune System Modulating and Anticancer Active Fractions from the Herbal Prescriptions

  • Hwang, Jong-Hyun;Jeong, Jae-Hyun;Yu, Kwang-Won
    • Food Science and Biotechnology
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    • v.18 no.2
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    • pp.323-329
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    • 2009
  • The prescriptions (DB-1-DB-5) were prepared with the active herbal medicines, Acanthopanax senticosus, Glycyrrhiza uralensis, Polygonatum odoratum, and Cichorium intybus. The most active crude polysaccharide fraction (DB-2-3), which was isolated through the fractionation of hot-water extract from DB-2, was significantly reduced by periodate oxidation (52.7 and 63.7%) on intestinal immune system modulating and anticancer activity. When DB-2-3 was further fractionated by column chromatographies, DB-2-3IIc-2 showed the most potent activities. In addition, DB-2-3IIc stimulated the proliferation of bone marrow cells via Peyer's patch in dose-dependent pattern by oral administration. The metastasis of colon 26-M3.1 lung carcinoma had significantly inhibited in mice fed DB-2-3IIc at 1 mg/mouse (43.8%). DB-2-3IIc-2 mainly contained uronic acid (46.1%) and 42.5% of neutral sugar with a small amount of protein (7.6%), and component sugar analysis also showed that DB-2-3IIc-2 was composed Ara, Gal, and GalA (molar ratio; 0.50:0.63:1.00). It may be suggested that activities of DB-2-3IIc-2 are resulted from pectic polysaccharides containing a polygalacturonan moiety with side chain of neutral sugars, such as Ara and Gal.

Isolation and Characterization of a Gene Encoding Hexokinase from Loquat (Eriobotrya japonica Lindl.)

  • Qin, Qiaoping;Zhang, Lanlan;Xu, Kai;Jiang, Li;Cheng, Longjun;Xu, Chuanmei;Cui, Yongyi
    • Horticultural Science & Technology
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    • v.30 no.3
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    • pp.243-249
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    • 2012
  • Hexokinase is the first enzyme in the hexose assimilation pathway; it acts as a sensor for plant sugar responses, and it is also important in determining the fruit sugar levels. The full-length cDNA of a hexokinase gene was isolated from loquat through reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends, which was designated as EjHXK1. EjHXK1 is 1,839 bp long and contains an entire open reading frame encoding 497 amino acids. The predicted protein of EjHXK1 shares 72%-81% similarity with other plant hexokinases. Phylogeny analysis indicated that EjHXK1 is closely related to maize and rice hexokinases. Transient expression of the 35S: EjHXK1-GFP fusion protein was observed on the cell membrane and cytoplasm. Real-time RT-PCR indicated that EjHXK1 is expressed in loquat leaves, stems, flowers, and fruits. EjHXK1 transcripts were higher during early fruit development, but decreases before maturation, which is consistent with hexokinase enzyme activity during fruit development and conducive for hexose accumulation in mature fruits. These results imply that EjHXK1 may play important roles in the regulation of sugar flux during fruit ripening.

Some Peroxysterols and Ceramides from "Phellinus ribis", a Korean Wild Mushroom

  • Moon, Dongcheul;Hwang, Kyunghwa;Choi, Kyuyeol;Choi, Dongcheol;Kim, Changsoo;Kim, Jaegil;Lee, Yongmoon;Zee, Okpyo
    • Analytical Science and Technology
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    • v.8 no.4
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    • pp.901-906
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    • 1995
  • Studies on the chemical constituents from a Korean wild mushroom, Phellinus ribis, were carried out. A triterpenoid, two peroxysterols, and a chlorobenzene compound were isolated from the hexane soluble fraction of the methanol extract of dried fruiting bodies of the basidomycetes. Those compounds identifed were 3-hydroxy-20(29)-lupen-28-oic acid (betulinic acid), 5,8-epidioxyergosta-6,22-dien-3-ol(ergosterol peroxide), 5,8-epidioxyergosta-6,9(11),22-trien-3-ol (dehydroperoxyergosterol), and 1,2,4,5-tetrachloro-3,6-dimethoxybenzene. Structural studies were carried out on molecular species of a ceramide and cerebroside isolated from the chloroform soluble fraction of the methanol extract. For ceramide, the major component fatty acids were a-hydroxy fatty acid isomers of $C_{22:00}{\sim}C_{25:00};$ the predominant long-chain bases were trihydroxy sphinganine of $C_{17}{\sim}C_{18}$. The structure of a cerebroside containing mono-sugar was assumed that the long-chain base was $C_{19:2}$ sphingadienine; the major fatty acids were $C_{16}{\sim}C_{15}$ ${\alpha}$-hydroxy fatty acid isomers.

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Change of Bacillus cereus Flavonoid O-Triglucosyltransferase Into Flavonoid O-Monoglucosyltransferase by Error-Prone Polymerase Chain Reaction

  • Jung, Na-Ri;Joe, Eun-Ji;Kim, Bong-Gyu;Ahn, Byoung-Chan;Park, Jun-Cheol;Chong, You-Hoon;Ahn, Joong-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.20 no.10
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    • pp.1393-1396
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    • 2010
  • The attachment of sugar to flavonoids enhances their solubility. Glycosylation is performed primarily by uridine diphosphate-dependent glycosyltransferases (UGTs). The UGT from Bacillus cereus, BcGT-1, transferred three glucose molecules into kaempferol. The structural analysis of BcGT-1 showed that its substrate binding site is wider than that of plant flavonoid monoglucosyltransferases. In order to create monoglucosyltransferase from BcGT-1, the error-prone polymerase chain reaction (PCR) was performed. We analyzed 150 clones. Among them, two mutants generated only kaempferol O-monoglucoside, albeit with reduced reactivity. Unexpectedly, the two mutants harbored mutations in the amino acids located outside of the active sites. Based on the modeled structure of BcGT-1, it was proposed that the local change in the secondary structure of BcGT-1 caused the alteration of triglucosyltransferase into monoglucosyltransferase.

Cloning and Characterization of Cyclohexanol Dehydrogenase Gene from Rhodococcus sp. TK6

  • CHOI JUN-HO;KIM TAE-KANG;KIM YOUNG-MOG;KIM WON-CHAN;JOO GIL-JAE;LEE KYEONG-YEOLL;RHEE IN-KOO
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1189-1196
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    • 2005
  • The cyclohexanol dehydrogenase (ChnA), produced by Rhodococcus sp. TK6, which is capable of growth on cyclohexanol as the sole carbon source, has been previously purified and characterized. However, the current study cloned the complete gene (chnA) for ChnA and its flanking regions using a combination of a polymerase chain reaction (PCR) based on the N-terminal amino acid sequence of the purified ChnA and plaque hybridization from a phage library of Rhodococcus sp. TK6. A sequence analysis of the 5,965-bp DNA fragment revealed five potential open reading frames (ORFs) designated as partial pte (phosphotriesterase), acs (acyl-CoA synthetase), scd (short chain dehydrogenase), stp (sugar transporter), and chnA (cyclohexanol dehydrogenase), respectively. The deduced amino acid sequence of the chnA gene exhibited a similarity of up to $53\%$ with members of the short-chain dehydrogenase/reductase (SDR) family. The chnA gene was expressed using the pET21 a(+) system in Escherichia coli. The activity of the expressed ChnA was then confirmed (13.6 U/mg of protein) and its properties investigated.

A Study on Sugars in Korean Sweet Rice Drink "Sikhye"(I) -Sugar Content and Its Composition- (식혜의 이소말토올리고당에 관한 연구(I) -정제 및 구조해석-)

  • 안용근
    • The Korean Journal of Food And Nutrition
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    • v.10 no.1
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    • pp.82-86
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    • 1997
  • A Korean traditional sweet rice drink "Sikye" was produced from the raw material of 20% of rice and 4% malt supplemented with 2l of tap water, by incubating the mixture at 6$0^{\circ}C$ for 7 hours. The product was found to contain 11.01% of maltose, 5.31% of isomaltooligosaccharides, 1.75% of maltotriose and 0.28% of glucose. Maltose, maltotriose and isomaltooligosaccharides in Sikye were seperated by ethanol (3 volume) precipitation repeated three times, followed by gel chromatography of Toyopearl HW-40S. 1H-NMR analysis revealed that the products of G2 and G3 size had only $\alpha$-1, 4-glucosidic linkage. but isomaltooligosaccharides showed both signal of $\alpha$-1, 4 and $\alpha$-1, 6-glucosidic linkage with its estimation ratio of 5:1. Isomaltooligosaccharides were hydrolyzed to produce maltooligosaccharide series from maltose to maltohexaose by pullulanase. These results, suggest that isomaltooligosaccharides were constructed by maltohexaose main chain with maltose or maltotriose and maltotetraose side chain.ide chain.

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Overproduction of Cellulose in Acetobacter xylinum KCCM 10100 Defective in GDP-Mannosyltransferase

  • Park Sang-Tae;Kim Eung-Bin;Kim Young-Min
    • Journal of Microbiology and Biotechnology
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    • v.16 no.6
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    • pp.961-964
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    • 2006
  • GDP-mannosyltransferase (GMT) is an enzyme responsible for the addition of a mannose to glucose ($\alpha$[1$\rightarrow$3]) during biosynthesis of the water-soluble branched polysaccharide acetan in Acefobacter species. In an effort to obtain a cellulose-overproducing bacterium, a mutant defective in GMT of Acetobacter xylinum KCCM 10100 was constructed by single crossover homologous recombination using part of the aceA gene encoding GMT amplified by polymerase chain reaction. The GMT-disrupted mutant produced 23% more cellulose, but 16% less water-soluble polysaccharide than those of the wild-type strain. Analysis of the sugar composition by gel permeation chromatography revealed that water-soluble polysaccharides produced by the GMT-defective mutant contained no mannose molecule.

Novel CRF1-receptor Antagonists from Pulsatilla koreana Root

  • Li, Wei;Noh, Hyojin;Lee, Sunghou;Lee, Min Ho;Lee, Eun Young;Kang, Sangjin;Kim, Young Ho
    • Natural Product Sciences
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    • v.20 no.4
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    • pp.281-284
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    • 2014
  • In this study, twenty-one oleanane-type triterpenoid saponins were isolated from a methanol extract of the roots of Pulsatilla koreana. Antagonistic activities were measured in these compounds by the aequorin based cellular functional assay system for the corticotropin releasing factor receptor (CRF1). Of them, compounds 7 - 10 showed the highest degree of CRF1 inhibition further at the concentration of $10{\mu}M$. Moreover, by the analysis based on the structure-activity relationship of isolated saponins, a sugar chain at C-3 and a carboxyl group at C-28, as well as a methyl group at C-23 seems to be key functional elements. To our knowledge, this is the first report on CRF1 inhibition of saponins from P. koreana.

Systematic Chirality Investigations of Zn-TLM binding Sites by 2D-NOESY Back-calculations

  • Kim, Daesung;Hoshik Won
    • Journal of the Korean Magnetic Resonance Society
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    • v.2 no.1
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    • pp.50-58
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    • 1998
  • The systematic chirality investigations were made on the basic of the fact that zinc-binding tallysomycin (ZnTLMA) could have chiral centers (Zn, NC3, C6) at possible 4-, 5-, and 6-coordination models. Although our NMR data exhibit that the ligation sites are ${\beta}$-aminoalanine, ${\beta}$-hydroxyhistidine, and pyrimidine moiety, all possible coordination modes were tested out to see what kind of chiralities on NC3-C6 are favorable to each coordination mode. Tests were also made that take into account the specific configuration of functional groups, including ${\beta}$-aminoalanine, sugar ring, and ${\beta}$-hydroxyhistidine. Tests were finally extended to zinc-water binding and specific conformational studies by introducing various hydrogen bonding networks associated with the propionamide side chain and the carbamide group of mannose. Results of systematic chirality investigations exhibit that the S-S configuration of NC3-C6 is favorable to all of coordination models, but the R-S configuration, if exists at all, should have internal strain on C6 chiral center.

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