• Korean Journal of Microbiology
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• v.46 no.4
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• pp.359-365
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• 2010

Fifty Actinobacteria strains were isolated from rhizosphere soil of Sasa borealis. In the course of screening for antibacterial activity against bacterial leaf spot of pepper (Xanthomonas axonopodis pv. vesicatoria) of isolates, 12 isolates showed strong antibiotic activity. Basis on the 16S rRNA gene sequence, they were belonging to Streptomyces cluster II. Strain JR-24 exhibited strong antibiotic activity against X. axonopodis pv. vesicatoria, had a minimum inhibitory concentration of 10 ${\mu}l$/disc. The strain JR-24 was most closely related to Streptomyces galbus $DSM40089^T$ (98.1%), Streptomyces longwoodensis $LMG20096^T$ (98%) and Streptomyces capoamus $JCM4734^T$ (97.8%). When assayed with the API 20NE and 50 CHE kit, it is positive for utilization of L-arabinose, D-fructose, D-glucose, D-galactose and hydrolysis of gelatin, protein, starch. The strains contained iso-$C_{14:0}$ (25.93%), iso-$C_{15:0}$ (10.13%), anteiso-$C_{15:0}$ (19.29%) and iso-$C_{16:0}$ (20.35%) as major fatty acids and MK-9 (H4), MK-9 (H6), and MK-9 (H8) as the isoprenoid quinone. Strain JR-24 was suggested new species of genus Streptomyces by nearest neighbors of genotypic relationships and phenotypic characterization. This study was important to microbial resources investigation for environment-friendly agriculture.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.6
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• pp.863-868
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• 2004

Lectin activities and chemical characteristics of Escherichia coli, Lactobacillus spp. and Bifidobacterium spp. originating from the porcine cecal mucosal layer were studied based on hemagglutination assay (HA) and hemagglutination inhibition assay (HIA). Although all the bacterial strains were able to agglutinate erythrocytes of porcine or rabbit origin, much higher HA titers were consistently observed for Lactobacillus spp. than for E. coli or for Bifidobacterium spp. A remarkable reduction in HA titers occurred by the treatment of E. coli and Lactobacillus spp. with protease or trypsin and of Bifidobacterium spp. with protease, trypsin or periodate. There were no significant effects on the HA titers of the three groups of bacteria after the treatment with lipase. Hemagglutination of E. coli was strongly inhibited by D (+)-mannose and D (+)-galactose; Lactobacillus spp. by $\alpha$-L-rhamnose and methyl-$\beta$-galactopyranoside; Bifidobacterium spp. by D (+)-alactose, $\alpha$-L-rhamnose, $\alpha$-L-fucose, L (+)-arabinose, D (+)-mannose, D (-)-fructose at a relatively low concentration (1.43 to 3.75 mg/ml). These results, combined with the enhanced HA activities of the three bacterial strains by modification of rabbit erythrocytes with neuraminidase and abolished HA activity of E. coli after treatment with $\beta$-galactosidase, indicate that it might be the glycoproteinous substances surrounding the surface of the bacterial cells that are responsible for the adhesions of these microorganisms by recognizing the specific receptors on the red blood cell.

• Journal of Applied Biological Chemistry
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• v.48 no.1
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• pp.7-10
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• 2005

Trichoderma sp. SY most effectively produces an extracellular ${\gamma}$-L-arabinofuranosidase (AF) using arabinose as a carbon source. AF grown on cellulose as a carbon source was purified 28-fold with 4.4% yield by DEAE exchange and HQ/20 cation exchange chromatographies The purified enzyme was found to be homogeneous on SDS-PAGE with molecular weight of 89 kDa. It exhibited a high level of activity with p-nitrophenyl ${\alpha}$-L-arabinofuranoside, showing $K_m$ and $V_{max}$ values of $0.15\;{\mu}M$ and $239.85U{\cdot}mg^{-1}$, respectively and did not require any metal ion for activity. It also released p-nitrophenol from p-nitrophenol conjugated ${\beta}$-D-xylopyranoside, and ${\beta}$-D-galactopyranoside not from ${\beta}$-D-glucopyranoside.

• Microbiology and Biotechnology Letters
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• v.22 no.6
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• pp.607-613
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• 1994

The Bacillus stearothermophilus arfI gene encoding a-arabinofuranosidase was isolated from the genomic library, cloned into pBR322, and subsequently transferred into the Escherichia coli HB101. The recombinant E. coli was selected from approximately 10,000 transformants screened by making use of its ability to produce a yellow pigment around the colony on the selective medium supplemented with p-nitrophenyl-$\alpha$-L-arabinofuranoside (pNPAf), a chromogenic substrate. The functional clone was found to harbor a recombinant plasmid, pKMG11 with an insertion of about 5 kb derived from the B. stearothermophilus chromosomal DNA. Identity of the arfI gene on the insert DNA was confirmed by a zymogram with 4-methylumbelliferyl-$\alpha$-L-arabinofuranoside as the enzyme substrate. The $\alpha$-arabinofuranosidase from the recombinant E. coli strain showed very high substrate specificity; the enzyme displayed high activity only with pNPAf among many other p- or $o$-nitrophenyl derivatives of several sugars, and acted only on arabinoxylan among various natural arabinose containing polysaccharides tested.

• Korean Journal of Food Science and Technology
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• v.34 no.5
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• pp.879-884
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• 2002

Cold and hot water fractions of Diospyros kaki were screened to determine its anti-complementary activity. Flour of Diospyros kaki leaf (250 g) was boiled at $100^{\circ}C$ for 3 h and passed through a membrane of 10 kDa molecular weight (DK-0). DK-0 was precipitated with ethanol and refluxed with methanol to obtain the crude polysaccharide (DKC). DKC-1 was isolated by ion exchange chromatography on DEAE-Toyopearl 650C, and DKC-1c was purified from DKC-1 by size exclusion chromatography on Bio gel P-60. The anti-complementary activities of DKC-1c at $1000\;{\mu}g/mL$ were 85.4 and 61.1% via whole and alternative pathways, respectively. DKC-1c was determined as a neutral polysaccharide composed of glucose (29.0 mol.%), arabinose (24.3 mol.%), and galactose (16.2 mol.%) with the molecular weight of 66.6 kDa. Results of agarose gel immunoelectrophoresis revealed DKC-1c, as a complement activator, cleaved C3 into C3a and C3b via both pathways.

• Journal of Microbiology and Biotechnology
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• v.17 no.3
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• pp.481-489
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• 2007

[ $\alpha$ ]-L-Arabinofuranosidases (AFases; EC 3.2.1.55) are exo-type enzymes, which hydrolyze terminal nonreducing arabinose residues from various polysaccharides such as arabinan and arabinoxylan. Genome-wide BLAST search showed that various bacterial strains possess the putative AFase genes with well-conserved motif sequences at the nucleotide and amino acid sequence levels. In this study, two sets of degenerate PCR primers were designed and tested to detect putative AFase genes, based on their three highly conserved amino acid blocks (PGGNFV, GNEMDG; and DEWNVW). Among 20 Bacillus-associated species, 13 species were revealed to have putative AFase genes in their genome and they share over 67% of amino acid identities with each other. Based on the partial sequence obtained from an isolate, an AFase from Geobacillus sp. was cloned and expressed in E. coli. Enzymatic characterization has verified that the resulting enzyme corresponds to a typical AFase. Accordingly, degenerate PCR primers developed in this work can be used for fast, easy, and specific detection and isolation of putative AFase genes from bacterial cells.

• Food Science and Biotechnology
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• v.17 no.5
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• pp.990-995
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• 2008

A thermostable $\alpha$-L-arabinofuranosidases ($\alpha$-L-AFase) is an industrially important enzyme for recovery of L-arabinose from hemicellulose. The recombinant $\alpha$-L-AFase from Thermotoga maritima was expressed in Escherichia coli by using a constitutive pHCE or an inducible pRSET vectors. In batch fermentation, the constitutive expression system resulted in slightly faster growth rate (0.78 vs. 0.74/hr) but lower enzyme activity (2,553 vs. 3,723 units/L) than those of the induction system. When fed-batch fermentation was performed, biomass and enzyme activity reached the highest levels of 36 g/L and 9,152 units/L, respectively. The fed batch cultures performed superior results than batch culture in terms of biomass yield (4.62-5.42 folds) and enzyme synthesis (3.39-4.00 folds). In addition, the fed-batch induction strategy at high cell density resulted in the best productivity in cell growth as well as enzyme activity rather than the induction method at low cell density or the constitutive expression.

• The Korean Journal of Food And Nutrition
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• v.30 no.6
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• pp.1140-1148
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• 2017

For the purpose of developing new immunomodulatory agents from broccoli, ethanol extract (BCEE), hot water extract (BCHW), and crude polysaccharide (BCCP) were isolated from broccoli, and their immunomodulatory activities and chemical properties were examined. In the in vitro cytotoxicity analysis, BCHW and BCCP did not affect the growth of tumor cells and normal cells. Murine peritoneal macrophages stimulated with BCCP showed higher production of IL-6, IL-12, and $TNF-{\alpha}$ cytokines than those stimulated with BCHW. Also, BCHW and BCCP did not show proliferation of splenic lymphocytes. In the in vitro assay for intestinal immunomodulatory activities, only BCCP enhanced GM-CSF secretion and the bone marrow cell-proliferating activity via cells in Peyer's patches at $1,000{\mu}g/mL$. Also, BCHW mainly contained 33.7% neutral sugars, such as arabinose, glucose, and galactose, and 30.7% uronic acid, and BCCP consisted of 42.6% neutral sugars, including arabinose, galactose, and glucose, and 50.5% uronic acid. The above results lead us to conclude that crude polysaccharide (BCCP) isolated from broccoli causes considerably high cytokine production in peritoneal macrophages and bone marrow cell proliferation, and the polysaccharide extraction process is indispensable for separation of new immunomodulatory agents from broccoli.

• Applied Biological Chemistry
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• v.39 no.2
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• pp.159-164
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• 1996

Screening of anticoagulant activity was conducted for the hot water extracts of 73 kinds of medicinal herbs, 41 kinds of Korean edible plants, and 5 kinds of sea weeds using plasma recalcification test(Tr). In the first screening several extracts of the plants, Alisma calndiculatum, Corydalis ternata Panax notoginseng, Allium sativum, Ganderma luidum, Codium fragile, showed high activities. When the plants were reextracted with various solvent conditions, acidic water extracts of Codium fragile showed the highest activity in APTT. A crude polysaccharide fraction(CF-1) was prepared by methanol reflux, ethanol precipitation, dialysis and Iyophilization of the acid extracts. CF-1 comprised 80.8% total sugar consisting of arabinose, galactose and glucose as the main monomers, 8.7% protein, and 13.3% sulfate. The anticoagulant activity of CF-1 was not changed by pronase digestion, but decreased by periodate oxidation, and this indicated that the anticoagulant activity was attributed to the polysaccharide portion.

• Microbiology and Biotechnology Letters
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• v.35 no.3
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• pp.191-195
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• 2007

Four bacteria producing viscous exopolysaccharides (EPSs) were isolated from Korean fermented vegetables (Cucumber kimchi, Young radish kimchi, Green onion kimchi) using a selection medium intended for isolating bacteria with tannin-degrading activity. They were identified phylogenetically by 16S rDNA sequence analysis and found to be very close to Enterobacter cowan ii, Escherichia senegalensis, Enterobacter asburiae, and Enterobacter ludwigii. Strain CK31, the most efficient EPS-producer, produced a heteropolysaccharide with an approximate molecular weight of 420 kDa. The neutral sugar fraction of the EPS was composed of rhamnose, fucose, arabinose, mannose, galactose, and glucose.