• Korean Journal of Pharmacognosy
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• v.28 no.2
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• pp.65-71
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• 1997

This study was carried out to purify and to characterize various bioactive material acemannan from Aloe vera. Purified acemannan was mannose (67%) and acetyl group (23%), and the rest of glucose was galactose that consists of long chain polydispered beta-(1, 4) linked mannan polymers. The sugar and acetyl group in the molecule were linked by molar ratio of 3 : 1. This polysaccharide from Aloe vera may provide functional flood and potential drug source with antiviral and immunomodulating properties.

• Microbiology and Biotechnology Letters
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• v.22 no.4
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• pp.373-381
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• 1994

An actinomycete strain, HSL-613 was isolated -from soil and identified by International Streptomyces Project (ISP) and chemotaxonomic methods. The spore chain of the strain HSL-613 appears in a spiral shape, and its spores are spherical shape with smooth surface. The cell wall contains LL-diaminopimelic acid (DAP). Menaquinone MK-9 (H$_{6}$, H$_{8}$) and iso- and anteiso-branched fatty acids were detected from whole cell extract. Sugars identified from whole cell extract include galactose, glucose, mannose and ribose, which are distinct from general sugar patterns of Streptomyces. Average G+C content in the chromosome is 59%. 5S rRNA of HSL-613 consists of 120 nucleotides as determined by comparing with that of a type strain Streptomyces griseus subsp. KCTC 9080. Through morphological, physiological, and chemical characterization, HSL-613 was identified and named as Streptomyces sp. HSL-613.

• Food Science and Preservation
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• v.15 no.5
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• pp.617-621
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• 2008

Sequential passes through $Sephadex^{TM}$ columns were used to select phages that displays ligands for dextran ($\alpha$-1,6 linked linear chains) from a phage antibody library. Those phages that bound to the $Sephadex^{TM}$ in each iteration were replicated in E. coli. A phage preparation isolated on the third round selection produced 5.4 nephelos turbidity units (NTU) in a dextran specific immunonephelometric assay, a 2.2 fold higher value than the phage preparation from the first round selection. This phage gave $72\;{\pm}\;10$ normalized intensity (N.I.) in a dip-stick assay against high molecular size dextran (T2000, $2\;{\times}\;10^6) and significantly lower color ($30\;{\pm}\;6$ N.I.) against low molecular size dextran (T10, $10^4$). The presence of an Fab insert in each of these phages was confirmed using a $\beta-galactosidase linked assay and polymerase chain reaction.

• Journal of Ginseng Research
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• v.44 no.4
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• pp.570-579
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• 2020

Background: Many researchers reported that the various immune activities of red ginseng are due to acid polysaccharides. But, the exact structural characteristics of the acidic polysaccharide in red ginseng have not been fully elucidated. Therefore, we isolated the acidic polysaccharide from red ginseng and characterized the structural property of the active moiety of this polysaccharide, which contributes to the immunostimulatory activity of red ginseng. Methods: A polysaccharide (RGP-AP-I) was purified from red ginseng via size-exclusion chromatography using Sephadex G-100. Immunostimulatary activity of RGP-AP-I was investigated via anti-complementory and macrophage stimulatory activity. The structure of RGP-AP-I was characterized by HPLC, sugar composition, β-glucosyl Yariv reagent and methylation analysis. Results: Peritoneal macrophages stimulated using RGP-AP-I significantly augmented the production of various cytokines such as interleukin (IL)-6, IL-12, and tumor necrosis factor (TNF)-α. The primary structure of RGP-AP-I was elucidated by assessing its sugar composition and methylation analysis. RGP-AP-I is a 96 kDa acidic polysaccharide, and comprises nine different monosaccharides, which mainly include sugars such as rhamnose (Rha, 9.5%), galacturonic acid (GalA, 18.4%), galactose (Gal, 30.4%), and arabinose (Ara, 35.0%). RGP-AP-I exhibited an considerable reaction with the β-glucosyl Yariv reagent, revealing the presence of arabino-β-3,6-galactan. Methylation analysis indicated that RGP-AP-I comprises 21 different glycosyl linkages, such as 3-, 4-, 6- and 3,6-linked Galp; 5-linked Araf; 2,4-linked Rhap; and 4-linked GalAp, which are characteristics of rhamnogalacturonan I (RG-I). Conclusion: we assumed that the immunostimulatory activity of RGP-AP-I may be due to the RG-I structure, which comprises a main chain with a repeating linkage unit, [→2)-Rhap-(1→4)-GalAp-(1→] and three groups of side chains such as (1→5)-linked arabinan, (1→4)-linked galactan, and arabino-β-3,6-galactan, which branch at the C(O)4 positions of Rha residues in the main chain of RGP-AP-I.

• Journal of Korean Medicine for Obesity Research
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• v.18 no.2
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• pp.106-114
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• 2018

Objectives: We investigated about the microbial properties and changes in the efficacy of the Codonopsis lanceolata (CL) by natural fermentation. Methods: CL was fermented for four weeks in a well-ventilated place with 2.5% salt. pH, total sugar, total polyphenol, and total flavonoid were measured to determine fermentation characteristics according to fermentation period and salt treatment. Polymerase chain reaction denaturing gradient gel electrophoresis and random amplification of polymorphic DNA-polymerase chain reaction were carried out for microbial analysis during fermentation. In addition, HepG2 cell was cultured to check the lipid accumulation through oil red O staining and the glucose uptake was analyzed by measuring the 2-NBDG at C2C12 cell. Results: The pH level and the total sugar decreased with the CL fermentation. Total polyphenol and flavonoid increased after CL fermentation. It was confirmed that Leuconostoc mesenteroides were maintained continuously during fermentation. In the salt treatment CL, there was a sharp increase in Rahnella aquatilis. Lactobacillus plantarum matrix was observed in fermented CL. In addition, Lactococcus lactis, Weissella koreensis, R. aquatilis, L. plantarum, Leu. mesenteroides have been added to the salt treatment. Glucose uptake were significantly increased after fermentation with salt for four weeks. Lipid accumulation in the HepG2 cells was observed that there was difference (P<0.01) between free fatty acid group (100%) and decreased 4 weeks after fermentation (90.38%) at $800{\mu}g/mL$. Conclusions: Total polyphenol and flavonoid were increased after CL fermentation. Especially, percentage of the glucose uptake and lipid accumulation inhibition increased in CL fermentation with salt. It is expected that fermentation of salt treated CL will be more effective in diabetes and fatty liver.

• Journal of Microbiology and Biotechnology
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• v.19 no.9
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• pp.932-945
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• 2009

Several species of Gram-positive bacteria have cell wall peptidoglycan (syn. murein) in which not all of the sugar moieties are N-acetylated. This has recently been shown to be a secondary effect, caused by the action of a peptidoglycan N-acetylglucosamine deacetylase. We have found that the opportunistic pathogen Listeria monocytogenes is unusual in having three enzymes with such activity, two of which remain in the cytoplasm. Here, we examine the enzyme (PgdA) that crosses the cytoplasmic membrane and is localized in the cell wall. We purified a hexa-His-tagged form of PgdA to study its activity and constructed a mutant devoid of functional Lmo0415 (PgdA) protein. L. monocytogenes PgdA protein exhibited peptidoglycan N-acetylglucosamine deacetylase activity with natural substrates (peptidoglycan) from both L. monocytogenes and Escherichia coli as well as the peptidoglycan sugar chain component N-acetylglucosamine, but not with N-acetylmuramic acid. As was reported recently [6], inactivation of the structural gene was not lethal for L. monocytogenes nor did it affect growth rate or morphology of the cells. However, the pgdA mutant was more prone to autolysis induced by such agents as Triton X-100 and EDTA, and is more susceptible to the cationic antimicrobial peptides (CAMP) lysozyme and mutanolysin, using either peptidoglycan muramidases or autolysis-inducing agents. The pgdA mutant was also slightly more susceptible than the wild-type strain to the action of certain beta-lactam antibiotics. Our results indicate that protein PgdA plays a protective physiological role for listerial cells.

• Journal of the Korean Wood Science and Technology
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• v.24 no.3
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• pp.28-36
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• 1996

This experiment was carried out to elucidate the sugar composition of polysaccharides and the structural features of water-soluble polysaccharides(WSP) isolated from Korean oak mistletoe, Loranthus yadoriki Sieb. The 48-hours ball-milled meals of extractive-free dried mistletoe sawdusts were extracted with distilled water for $24hrs{\times}2$ at room temperature. The extracts poured into 95% ethyl alcohol to precipitate. The separated precipitate of WSP, in form of yellowish white powder by lyophilization, was fractionated into four subfractions of WSP-1, WSP-2, WSP-3 and WSP-4 by anion exchange chromatography on DEAE-cellulose column. The sugar composition of WSPs was analyzed by GLC in form of their glycitol acetates, and the structure of polysaccharides in Fractions WSP-1 and WSP-2 was determined by FT-IR and GC-MS after methylation through and acetylation. The sugars of WSPs from Korean oak mistletoe, Loranthus yadoriki, are majorly arabinose and galactose in stem, galactose in leaves very high in content and showed difference in composition and monomeric units between stems and leaves. D-galactose, D-glucose and L-arabinose are the simple sugars consisting of polysaccharides in WSP-1. ($1{\rightarrow}3$)-Linked galactan is the bakcbone with side chain of ($1{\rightarrow}5$)- -L-arabinofuranosyl residues and ($1{\rightarrow}6$)- -D-galactopyranosyl residues, and ($1{\rightarrow}4$)-linked glucan also presents. ($1{\rightarrow}4$)-Linked rhamnogalacturonan and ($1{\rightarrow}4$)- and ($1{\rightarrow}3$)-linked galactan present in WSP-2.

• Proceedings of the Botanical Society of Korea Conference
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• 1999.07a
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• pp.63-67
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• 1999

We have isolated more than a dozen cDNA clones corresponding to genes that were expressed in Arabidopsis leaves when they were kept in the dark. The nucleotide sequence analysis showed that some of the clones encoded proteins with significant homology to $\beta$-glucosidase (din2), branched-chain $\alpha$-keto acid dehydrogenase subunit E1$\beta$(din3), and another subunit E2 (din4), yeast RAD23 (din5), asparagine synthetase (din6), pre-mRNA splicing factor SRp35 (din7), phosphomannose isomerase (din9), seed imbibition protein (din10), and 2-oxoacid-dependent oxidase (din11). Accumulation of transcripts from din3,4,6 and 10 occurred rapidly after the plants were transferred to darkness. Transcripts from din2,9, and 11 could be detected only after 24 h of dark treatment. Inhibition of photo-synthesis by DCMU strongly induced the accumulation of transcripts from those genes, and application of sucrose to detached leaves suppressed the accumulation both in the dark and by DCMU. These observations indicate that expression of the genes is caused by sugar starvation resulted from the cessation of photosynthesis. We further showed that din2-encoded protein also accumulated in senescing leaves. Given these results, possible roles of din genes in leaves in the dark and senescing leaves are discussed.

• Journal of Microbiology and Biotechnology
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• v.23 no.2
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• pp.137-143
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• 2013

The diversity and distribution of methanogenic archaea in a four-compartment anaerobic baffled reactor (ABR) treating sugar refinery wastewater were investigated by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). At an organic loading rate of 5.33 kg $COD/m^3{\cdot}day$, the ABR could perform steadily with the mean chemical oxygen demand (COD) removal of 94.8% and the specific $CH_4$ yield of 0.21 l/g $COD_{removed}$. The $CH_4$ content in the biogas was increased along the compartments, whereas the percentage of $H_2$ was decreased, indicating the distribution characteristics of the methanogens occurred longitudinally down the ABR. A high phylogenetic and ecological diversity of methanogens was found in the ABR, and all the detected methanogens were classified into six groups, including Methanomicrobiales, Methanosarcinales, Methanobacteriales, Crenarchaeota, Arc I, and Unidentified. Among the methanogenic population, the acid-tolerant hydrogenotrophic methanogens including Methanoregula and Methanosphaerula dominated the first two compartments. In the last two compartments, the dominant methanogenic population was Methanosaeta, which was the major acetate oxidizer under methanogenic conditions and could promote the formation of granular sludge. The distribution of the hydrogenotrophic (acid-tolerant) and acetotrophic methanogens in sequence along the compartments allowed the ABR to perform more efficiently and steadily.

• Journal of Microbiology and Biotechnology
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• v.31 no.8
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• pp.1123-1133
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• 2021

Biodegradation is the key process involved in natural lignocellulose biotransformation and utilization. Microbial consortia represent promising candidates for applications in lignocellulose conversion strategies for biofuel production; however, cooperation among the enzymes and the labor division of microbes in the microbial consortia remains unclear. In this study, metagenomic analysis was performed to reveal the community structure and extremozyme systems of a lignocellulolytic microbial consortium, TMC7. The taxonomic affiliation of TMC7 metagenome included members of the genera Ruminiclostridium (42.85%), Thermoanaerobacterium (18.41%), Geobacillus (10.44%), unclassified_f__Bacillaceae (7.48%), Aeribacillus (2.65%), Symbiobacterium (2.47%), Desulfotomaculum (2.33%), Caldibacillus (1.56%), Clostridium (1.26%), and others (10.55%). The carbohydrate-active enzyme annotation revealed that TMC7 encoded a broad array of enzymes responsible for cellulose and hemicellulose degradation. Ten glycoside hydrolases (GHs) endoglucanase, 4 GHs exoglucanase, and 6 GHs β-glucosidase were identified for cellulose degradation; 6 GHs endo-β-1,4-xylanase, 9 GHs β-xylosidase, and 3 GHs β-mannanase were identified for degradation of the hemicellulose main chain; 6 GHs arabinofuranosidase, 2 GHs α-mannosidase, 11 GHs galactosidase, 3 GHs α-rhamnosidase, and 4 GHs α-fucosidase were identified as xylan debranching enzymes. Furthermore, by introducing a factor named as the contribution coefficient, we found that Ruminiclostridium and Thermoanaerobacterium may be the dominant contributors, whereas Symbiobacterium and Desulfotomaculum may serve as "sugar cheaters" in lignocellulose degradation by TMC7. Our findings provide mechanistic profiles of an array of enzymes that degrade complex lignocellulosic biomass in the microbial consortium TMC7 and provide a promising approach for studying the potential contribution of microbes in microbial consortia.