• Microbiology and Biotechnology Letters
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• v.43 no.4
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• pp.307-313
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• 2015

Unpolished rice (UR) is considered to be a healthy alternative to white rice when coping with chronic diseases. In the present study, the fermented slurry of unpolished rice (FSUR) was evaluated with respect to its antimicrobial activities and biochemical characteristics, including the quantities of sugar, total soluble sugar, organic acids, free amino acids, pH, and physiological activity. The antimicrobial efficiency of FSUR was assessed using the paper disc-agar diffusion method. FSUR exhibited strong antimicrobial activity against six pathogenic bacterial strains (Staphylococcus aureus, Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, and Yersinia enterocolitica) and two fermentation strains (Gluconacetobacter intermedius and Lodderomyces elongisporus). The antimicrobial activity of FSUR was higher than the commercial antibiotics, carbenicillin ($50{\mu}g/ml$) and tetracycline ($50{\mu}g/ml$) against S. aureus, E. coli, L. monocytogenes, P. aeruginosa, S. typhimurium, Y. enterocolitica, and L. elongisporus. Also FSUR had a high antioxidant activity. The microorganisms were isolated from FSUR using tryptic soy broth and yeast extract-peptone-dextrose agar media. The isolated microorganisms were characterized using physiological and biochemical analyses as well as by 16S rRNA gene sequencing and phylogenic analysis. 16S rRNA gene sequence analysis showed that the isolated microorganisms had a high similarity to G. intermedius, Lactobacillus casei, Lactobacillus plantarum, and Acetobacter peroxydans.

• Journal of Microbiology and Biotechnology
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• v.16 no.10
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• pp.1613-1621
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• 2006

Isolation and identification of algal lytic bacteria were carried out. Nine strains of algal lytic bacteria were isolated by the double-layer method using Anabaena flos-aquae as a sole nutrient. The isolate, AFK-13, showing the highest algal lytic activity was identified as Sinorhizobium kostiense based on the l6S rDNA sequence. The algal lytic experiments of the culture supernatants of AFK-13 demonstrated that the bacterial cell growth reached a maximum at 36-h culture, but the supernatant of 72-h culture exhibited the highest activity. Components among the extracellular products in the crude enzyme of the supernatant from S. kostiense AFK-13 culture were responsible for degradation of cell walls of Anabaena flos-aquae. Algal lytic assay tests of the culture supernatants suggest that the main substances for algal lytic activity could be proteinaceous. The activity of glucosidase was observed highly by polysaccharolytic analysis using the crude enzyme from S. kostiense AFK-13, whereas activities of galactosidase, mannosidase, rhamnosidase, and arabinosidase were also detected in low levels. The molecular weights (MW) of ${\alpha}-\;and\;{\beta}$-glucosidases were estimated to be approximately 50-100 kDa by the ultrafiltration method.

• Food Science of Animal Resources
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• v.32 no.5
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• pp.635-643
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• 2012

As an appraisal for the application of a new starter culture, more than 200 lactic acid bacteria strains were isolated from raw milk and healthy human feces. The strains showing excellent growth and acid production ability in 10% skim milk media were selected and identified as Lactobacillus casei based on the results of their API carbohydrate fermentation patterns, as well as 16S rDNA sequence analysis. To assess the effect of L. casei strains on irritable bowel disease (IBD), the inhibitory effect of the selected strains against the nitric oxide (NO) production of lipopolysaccharide (LPS)-stimulated RAW 264.7 cells was measured. Among the tested L. casei strains, L. casei MCL was observed to have the greatest NO inhibitory activity. Additionally, L. casei MCL was found to inhibit mRNA expression of pro-inflammatory cytokines (interleukin-$1{\beta}$, IL-6, TNF-${\alpha}$), as well as cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) involved in pathophysiologic processes such as inflammation. The mRNA expression of anti-inflammatory cytokines, including IL-10 and transforming growth factor-$1{\beta}$ (TGF-${\beta}$) of L. casei MCL, was confirmed using quantitative real-time PCR. In conclusion, L. casei MCL showed decreases in the expression of pro-inflammatory cytokines and up-regulated expression of the anti-inflammatory cytokine.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.1
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• pp.35-43
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• 2015

In this study, hot water extract from sea buckthorn (Hippophae rhamnoides L.) leaves fermented with Hericium erinaceum mycelium (SBT-HE) was assessed for protection against oxidative modification of biological macromolecules and cell death. Antioxidant activity of SBT-HE was evaluated based on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical, and peroxyl radical scavenging assays. SBT-HE showed 65.06% DPPH radical scavenging activity at $500{\mu}g/mL$, 98.83% ABTS radical scavenging activity at $50{\mu}g/mL$, and 44.03% peroxyl radical scavenging activity at $100{\mu}g/mL$. SBT-HE significantly inhibited DNA strand breakage induced by peroxyl radical. SBT-HE also prevented peroxyl radical-mediated human serum albumin modification. SBT-HE effectively inhibited $H_2O_2$-induced cell death and significantly increased cell survival by 21.59% at $100{\mu}g/mL$. SBT-HE also reduced intracellular reactive oxygen species levels in $H_2O_2$-treated cells. The results suggest that SBT-HE can contribute to antioxidant activity and protect cells from oxidative stress-induced cell injury.

• Microbiology and Biotechnology Letters
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• v.36 no.4
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• pp.268-275
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• 2008

In order to develop the multi-functional rhizobacteria that can exert positive effect on the growth of plants growing in the coastal sand dune located along East Coast of Korea, rhizospheral bacteria of 11 different plants from this area were isolated 1,330 rhizobacteria. Among these, 23 strains were able to produce auxin and had spectrum of antagonism toward various phytopathogenic microbes. To know the mechanism of this antifungal activity, these 23 strains were subjected to further analyses; 19 strains of these produced siderophore as determined by color reaction on CAS-blue plate, 4 strains produced antifungal cellulase as judged by color change on CMC-Congo red plate, 17 strains were able to utilized insoluble phosphate salts, also determined by clear zone formation on PVK medium. Identification of the strain was assigned to all 23 strains by l6s rDNA sequence analysed, and all were identified to be in the genus of Bacillus and Pseudomonas. One strain of these, denoted Pseudomonas fluorescens IB4-14, showed ACC deaminase activity which is known to be involved in the resistance of environmental stress such as salt and drought. Also, P. fluorescens IB4-l4 showed the germination stimulation and roots growth promoting activity on the in vivo assay of Lysimachia mauritiana Lam. (spoonleaf yellow loosestrife).

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.4
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• pp.296-303
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• 2005

A strain named as HJ35 was isolated from the skin of sixty-five men and fourteen women for acne therapy, in order to find an effective antimicrobial agent against Propionibacterium acnes. Isolate HJ35 was identified as Enterococcus faecium based on 16 rDNA sequence and produced enterocin HJ35 having antimicrobial activities against most lactic acid bacteria, En­terococcus spp., Staphylococcus aureus, S. epidermidis, Clostridium perfringens, some bacilli, Mi­crococcus flavus, Listeria monocytogenes, L. ivanovii, Escherichia coli, Pseudomonas fluorescens and Propionibacterium acnes, in the modified well diffusion method. Especially, enterocin HJ35 showed a bactericidal activity against Propionibacterium acnes P1. The antimicrobial activity of enterocin HJ35 was disappeared completely with the use of protease XIV. But enterocin HJ35 activity is very stable at high temperature (up to $100^{\circ}C$ for 30 min), in wide range of pH (3.0${\~}$9.0), and by treatment with organic solvents. The apparent molecular mass of enterocin HJ35 was estimated to be approximately 4${\~}$4.5 kDa on detection of its bactericidal activity after SDS-PAGE. In batch fermentation of E. faecium HJ35, enterocin HJ35 was produced at the mid­log growth phase, and its maximum production was obtained up to 2,300 AU/mL at the late stationary phase. By employing fed-batch fermentation, the enhanced production of enterocin HJ35 was achieved up to 12,800 AU/mL by feeding with 10 g/L glucose or 6 g/L lactate.

• KSBB Journal
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• v.26 no.6
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• pp.552-556
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• 2011

A novel agar-degrading bacterium mbrc-1 was isolated from seashore of Kyungpo at Gangwon province and cultured in marine broth 2216 medium. Isolated bacterium mbrc-1 was named as Flammeovirga sp. mbrc-1 based on the 16S rDNA sequence. Its agarase showed maximum activity of 923 units/L at pH 7.0 and $45^{\circ}C$ and sustained 90% remaining activity after exposed to $45^{\circ}C$ for 2 hours. The enzyme hydrolyzed agarose to yield neoagarohexaose (18.5%), neoagarotetraose (38%) and neoagarobiose (43.5%), indicating that the enzyme is ${\beta}$-agarase. Thus, isolated bacterium and its ${\beta}$-agarase would be useful for the industrial production of neoagarotetraose and neoagarobiose.

• Journal of Genetic Medicine
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• v.4 no.1
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• pp.72-79
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• 2007

Purpose : Glycogen storage disease type III (GSD-III) is a rare autosomal recessive disorder of glycogen metabolism. The affected enzyme, amylo-1,6-glucosidase, 4-alpha-glucanotransferase (AGL, glycogen debranching enzyme), is responsible for the debranching of the glycogen molecule during catabolism. The disease shows clinical and biochemical heterogeneity, reflecting genotype-phenotype heterogeneity among different patients. In this study, we aim at analyzing mutations of the AGL gene in three unrelated Korean GSD-III patients, and characterizing their clinical and laboratory findings. Methods : We characterized the clinical features of three unrelated Korean GSD-III patients by biochemical, histological and imaging studies. The 35 exons and part of exon-intron boundaries of AGL were analyzed by direct sequencing using genomic DNA extracted from the peripheral leukocytes of patients. Results : Diverse clinical features were observed in these patients including hepatomegaly (all patients), seizures (patient 2), grow th failure (patients 1 and 2), hyperlipidemia (patients 1 and 3), raised transaminase and creatine kinase concentrations (all patients), and mild cardiomyopathy (patient 2). Liver transplantation w as performed in patient 2 due to progressive hepatic fibrosis. A dministration of uncooked corn starch maintained normoglycemia and improved biochemical and growth profiles. DNA sequence analysis revealed mutations in 5 out of 6 alleles. Patient 1 was a compound heterozygote of c.1282 G>A (p.R428K) and c.1306delA (p.S603PfsX6), patient 2 had c.1510_1511insT (p.Y 504L fsX 10), and patient 3 had c.3416 T >C (p.L 1139P) and c.1735+1 G>T (p.Y 538_R578delfsX 4) mutations. A part from the p.R428K mutation, the 4 other substitutions identified w ere nov el. Conclusion : GSD-III patients display variable phenotypic characteristics resembling those of GSD-Ia. Molecular defects in the AGL gene of Korean GSD-III patients are genetically heterogeneous.

• The Journal of Natural Sciences
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• v.4
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• pp.103-142
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• 1991

These studies were carried out to select somatic hybrid using selectable marker genes of Nicotiana glauca transformed by NPTII gene and Solanum tuberosum transformed by T- DNA, and to study characteristics of transformant. The results are summarized as follows. 1. Crown gall tumors and hairy roots were formed on potato tuber disc infected by A. tumefaciens Ach5 and A. rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. 2. Callus formation from hairy root was prompted on the medium containing 2, 4 D 2mg/I with casein hydrolysate lg/l. 3. The survival ratio of crown gall tumor callus derived from potato increased on the medium containing the activated charcoal 0. 5-2. 0mg/I because of the preventions on the other hand, hairy roots were necrosis on the same medium. 4. Callus derived from hairy root were excellently grown for a short time by suspension culture on liquid medium containing 2, 4-D 2mg/I and casein hydrolysate lg/l. 5. The binary vector pGA643 was mobilized from E. coli MC1000 into wild type Agrobacteriurn tumefaciens Ach5, A. tumefaciens $A_4T$ and disarmed A. tuniefaciens LBA4404 using a triparental mating method with E. ccli HB1O1/pRK2013. Transconjugants were obtained on the minimal media containing tetracycline and kanamycin. pGA643 vectors were confirmed by electrophoresis on 0.7% agarose gel. 6. Kanamycin resistant calli were selected on the media supplemented with 2, 4-D 0.5mg/1 and kanamycin $100\mug$/ml after co- cultivating with tobacco stem explants and A. tumefaciens LBA4404/pGA643, and selected calli propagated on the same medium. 7. The multiple shoots were regenerated from kanamycin resistant calli on the MS medium containing BA 2mg/l. 8. Leaf segments of transformed shoot were able to grow vigorusly on the medium supplemented with high concentration of kanamycin $1000\mug$/ml. 9. Kanamycin resistant shoots were rooting and elongated on medium containing kanamycin $100\mug$/ml, but normal shoot were not. 10. For the production of protoplast from potato calli transformed by T-DNA and mesophyll tissue transformed by NPTII gene, the former was isolated in the enzyme mixture of 2.0% celluase Onozuka R-10, 1.0% dricelase, 1.0% macerozyme. and 0.5M mannitol, the latter was isolated in the enzyme mixture 1.0% Celluase Onozuka R-10, 0.3% macerozyme, and 0.7M mannitol. 11. The optimal concentrationn of mannitol in the enzyme mixture for high protoplast yield was 0.8M at both transformed tobacco mesophyll and potato callus. The viabilities of protoplast were shown above 90%, respectively. 12. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution. Cell walls were regenerated on hormone free media supplemented with kanamycin after 5 days, and colonies were observed after 4 weeks culture.

• Journal of Life Science
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• v.26 no.4
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• pp.453-459
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• 2016

Agarases are classified into α-agarase and β-agarase that produce agarooligosaccharides and neoagarooligosaccharides, respectively. Neoagarooligosaccharides have whitening effect of skin, delay of starch degradation, and inhibition of bacterial growth etc. Hence, the object of this study was to isolate a novel agarase producing marine bacterium and characterization of its β-agarase. A novel agar-degrading bacterium was isolated from seashore of Namhae at Gyeongnamprovine, Korea and purely cultured with Marine agar 2216 media. The isolated bacterium was identified as Simiduia sp. SH-4 after 16S rRNA gene sequencing. The enzymatic sample was obtained from culture media of Simiduia sp. SH-4. Enzymatic activity was highly increased from 20(30% relative activity) to 30℃ (100%) and decreased from 30 to 40℃(75%) and so more. Relative activity was 100% at pH 6 while those were about 91% and 59% at pH 5.0 and 7.0, respectively, meaning the enzyme possesses narrow optimal pH range. Hence, the enzyme exhibited the maximal activity with 120.4 units/l at pH 6.0 and 30℃ in 20 mM Tris-HCl buffer. Thin layer chromatography (TLC) analysis showed that Simiduia sp. SH-4 produces β-agarase, which hydrolyze agarose to produce biofunctional neoagarooligosaccharides such as neoagarotetraose and neoagarobiose. Hence, broad applications would be possible using Simiduia sp. SH-4 and its enzyme in the food industry, cosmetics and medical fields.