• Microbiology and Biotechnology Letters
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• v.40 no.1
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• pp.57-65
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• 2012

Antifungal compounds from Bacillus polyfermenticus CJ6 were purified using SPE, preparative HPLC, and reverse phase-HPLC. Antifungal compounds from B. polyfermenticus CJ6 were separated into three fractions (8, B, C) using preparative HPLC. LC/MS analysis of antifungal peaks suggested that B. polyfermenticus CJ6 produces lipopeptides; two kinds of iturin A ($C_{14}$, $C_{15}$), three kinds of surfactins ($C_{13}$, $C_{14}$, $C_{15}$), four kinds of fengycin A ($C_{14}$, $C_{15}$, $C_{16}$, $C_{17}$) and two kinds fengycin B ($C_{16}$, $C_{17}$). The antifungal activity of fraction 8, which was presumed as inturin A, was found to be stable after the pH, heat or proteolytic enzyme treatment, but it was unstable at 50-$70^{\circ}C$ for 24 hr. The antifungal activity of fraction B, which presumed as surfactins and fengycin A, was found to be stable after the heat treatment, but it was unstable in the pH 3.0 and after the protease (type I) or ${\alpha}$-chymotrypsin treatment. The antifungal activity of fraction C, which was presumed as fengycin A and B, was found to be stable in the pH 3.0-9.0 range and the heat treatment, but it was unstable with the treatment of protease (type I). The amino acid composition of the purified peaks 8-1 and 8-2 were Asx, Tyr, Gln, Pro, and Ser in a molar ratio of 3:1:1:1:1, which showed the same amino acid composition as iturin. From these results, we confirmed that antifungal compounds from B. polyfermenticus CJ6 most likely belonged to iturin A as well as surfactins and fengycins. As lipopeptides are known to act in a synergistic manner, the antifungal compounds from B. polyfermenticus CJ6 might have potential uses in biotechnology and biopharmaceutical applications.

• Journal of Dairy Science and Biotechnology
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• v.32 no.2
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• pp.131-139
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• 2014

The aim of this study was to manufacture Cutting-Gouda cheese and to investigate the change in physicochemical properties of Cutting-Gouda cheese made with Lactobacillus rhamnosus_p1. Lactic acid bacteria were isolated from Gouda cheese ripened for more than 1 year. They were identified as 2 strains of L. rhamnosus, Lactobacillus casei, Lactobacillus curvatus, and Staphylococcus saprophyticus by 16S rDNA sequencing and named L. rhamnosus_p1, L. casei_p2, L. curvatus_p3, L. rhamnosus_p4 and S. saprophyticus_p5. The proteolytic activities of isolated strains against casein were measured using prepared skim milk agar plates. L. rhamnosus_p1 showed the highest proteolytic activity. Cutting-Gouda cheese was made with L. rhamnosus_p1, and its physicochemical properties (moisture, protein, fat, ash and free amino acid content) were measured during ripening periods. Because of the modified atmosphere packaging ($N_2{^-}$), there was no change in moisture, protein, fat, and ash in the experimental group. The total amount of free amino acids in the control and experimental group gradually increased during ripening periods. The sensory evaluation showed that the experimental group was preferable to the control group. This result suggests that L. rhamnosus_p1 has potential to be developed as a new starter for Gouda cheese.

• Journal of Animal Science and Technology
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• v.46 no.5
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• pp.773-782
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• 2004

The objective of this work was to establish an approaching method for TMR feeding system in the farm situation by testing the effects of herd-mix feeding system on the performance of lactating cows. Fifty six Holstein cows were fed for the experimental period of 16 months. Prior to test the herd-mix feeding system, animals were kept on conventional feeding system for 4 months, separate feeding of forage with concentrate, then provided 3 types of herd-mix rations formulated by mean level of energy concentration requirement of higher 1/2 ranker in each herd for remaining 12 months to compare the effect on milk yield performance of animals with conventional feeding system by using a switch-over method. The herd-mix feeding system influenced substantially upon the improvements of milk yield(P<0.01) and milk fat percentage(P<0.05). In all of lactational ages, milk yield and milk fat were enhanced by the herd-mix feeding system. Especially, actual milk yield(AMY) and milk fat in the 1st lactating cows, and AMY and 4.0% fat corrected milk yield(FCM) in the 2nd lactating cows were increased signiticantly by the herd-mix feeding system(P<0.05). In the early and mid-stage of lactation, the herd-mix feeding system showed higher AMY and FCM compared with the conventional feeding system although the milk fat was not different. AMY, milk fat and FCM in the late-stage of lactation were increased generally by the herd-mix feeding system. Especially, milk fat and FCM obtained by the herd-mix feeding system were significantly higher than those by the conventional feeding system(P<0.01). The herd-mix feeding system showed a good type of milking curve with a higher persistency of FCM(93.24%) than in conventional one(92.69%). The income over feed cost with the herd-mix feeding system was lA-fold higher than that with the conventional feeding method. In conclusion, the results of this work suggest that the herd-mix feeding system based on a correct level of energy concentration of TMR and well-determined feed ingredients increases not only milk yield of dairy herd by enhancing the performance of cows in early to mid stage of lactation with improved milk persistency but also gross income of dairy farm.

• Journal of KIISE:Information Networking
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• v.30 no.3
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• pp.329-342
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• 2003

Most of the existing multicast routing protocols for ad-hoc networks do not take into account the efficiency of the protocol for the cases when there are large number of sources in the multicast group, resulting in either large overhead or poor data delivery ratio when the number of sources is large. In this paper, we propose a multicast routing protocol for ad-hoc networks, which particularly considers the scalability of the protocol in terms of the number of sources in the multicast groups. The proposed protocol designates a set of sources as the core sources. Each core source is a root of each tree that reaches all the destinations of the multicast group. The union of these trees constitutes the data delivery mesh, and each of the non-core sources finds the nearest core source in order to delegate its data delivery. For the efficient operation of the proposed protocol, it is important to have an appropriate number of core sources. Having too many of the core sources incurs excessive control and data packet overhead, whereas having too little of them results in a vulnerable and overloaded data delivery mesh. The data delivery mesh is optimally reconfigured through the periodic control message flooding from the core sources, whereas the connectivity of the mesh is maintained by a persistent local mesh recovery mechanism. The simulation results show that the proposed protocol achieves an efficient multicast communication with high data delivery ratio and low communication overhead compared with the other existing multicast routing protocols when there are multiple sources in the multicast group.

• Radiation Oncology Journal
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• v.19 no.3
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• pp.245-251
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• 2001

Prupose : The genes involved on the suppression or radiation-induced apoptosis by genistein in K562 leukemia cell line was investigated. Materials and methods : K562 cells in exponential growth phase were irradiated with a linear accelerator at room temperature. For X-ray irradiation and drug treatment, cultures were prepared at $2\times10^5\;cells/mL$. The cells were irradiated with 10 Gy (Clinac 1800C, Varian, USA), Stock solutions of herbimycin A (HMA, Calbiochem, UK) and genistein (Calbiochem, UK) were prepared in dimethylsulfoxide (DMSO, Sigma, UK). After incubation at $37^{\circ}C$ for 24 h, PCR-select cDNA subtractive hybridization, dot hybridization, DNA sequencing and Northern hybridization were examined. Results : Smad6 gene was identified from the differentially expressed genes in K562 cells incubated with genistein which had been selected by PCR-select cDNA subtractive hybridization. The mRNA expression of Smad6 in K562 cells incubated with genistein was also higher than control group by Northern hybridization analysis. Conclusion : We have shown that Smad6 involved on the suppression of radiation-induced apoptosis by genistein in K562 leukemia cell line. It is plausible that the relationship between Smad6 and the suppression of radiation-induced apoptosis is essential for treatment development based on molecular targeting designed to modify radiation-induced apoptosis.

• Korean Journal of Microbiology
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• v.42 no.2
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• pp.89-94
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• 2006

The purpose of this study is to develop the strain-specific PCR primers for the identification of prevotella inter-media ATCC 49046 which is frequently used in the pathogenesis studies of periodontitis. The Hind III-digested genomic DNA of P. intermedia ATCC 49046 were cloned by random cloning method. The specificity of cloned DNA fragments were determined by Southern blot analysis. The nucleotide sequence of cloned DNA probes was determined by chain termination method. The PCR primers were designed based on the nucleotide sequence of cloned DNA fragment. The data showed that Pig6 DNA probe were hybridized with the genomic DNA from P. intermedia strains (ATCC $25611^T$ and 49046) isolated from the Westerns, not the strains isolated from Koreans. The Pig6 DNA probe were consisted of 813 bp. Pig6-F3 and Pig6-R3 primers, designed base on the nucleotide Sequences Of Pig6 DNA Probe, were 3150 specific to the only both P. intermedia ATCC $25611^T$ and P. intermedia ATCC 49046. In the other hand, Pig6-60F and Pig6-770R primers were specific to the only P. intermedia ATCC 49046. The two PCR primer sets could detect as little as 4 pg of chromosomal DNA of P. intermedia. These results indicate that Pig6-60F and Pig6-770R primers have proven useful for the identification of P. intermedia ATCC 49046, especially with regard to the maintenance of the strain.

• Korean Journal of Microbiology
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• v.54 no.1
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• pp.60-68
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• 2018

A cold-active and alkaline serine protease (Pro21717) was partially purified from the Antarctic marine bacterium Pseudoalteromonas arctica PAMC 21717. On a zymogram gel containing skim milk, Pro21717 produced two distinct clear-zones of approximately 37 kDa (low intensity) and 74 kDa (high intensity). These were found to have identical N-terminal sequences, suggesting they arose from an identical precursor and that the 37 kDa protease might homodimerize to the more active 74 kDa form of the protein. Pro21717 displayed proteolytic activity at $0-40^{\circ}C$ (optimal temperature of $40^{\circ}C$) and maintained this activity at pH 5.0-10.0 (optimal pH of 9.0). Notably, relative activities of 30% and 45% were observed at $0^{\circ}C$ and $10^{\circ}C$, respectively, in comparison to the 100% activity observed at $40^{\circ}C$, and this enzyme showed a broad substrate range against synthetic peptides with a preference for proline in the cleavage reaction. Pro21717 activity was enhanced by $Cu^{2+}$ and remained stable in the presence of detergent surfactants (linear alkylbenzene sulfonate and sodium dodecyl sulfate) and other chemical components ($Na_2SO_4$ and metal ions, such as $Ba^{2+}$, $Mg^{2+}$, $Ca^{2+}$, $Zn^{2+}$, $Fe^{2+}$, $K^+$, and $Na^{2+}$), which are often included in commercial detergent formulations. These data indicate that the psychrophilic Pro21717 has properties comparable to the well-characterized mesophilic subtilisin Carlsberg, which is commercially produced by Novozymes as the trademark Alcalase. Thus it has the potential to be used as a new additive enzyme in laundry detergents that must work well in cold tap water below $15^{\circ}C$.

• Journal of Life Science
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• v.23 no.2
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• pp.167-174
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• 2013

Peroxiredoxin 6 (Prx 6) is a member of the thiol-specific antioxidant protein family, which may play a role in protection against oxidative stress and in regulating phospholipid turnover. The aim of this study was to determine whether a human Prx 6/Luc vector was stably expressed and responded to antioxidants in a lung cell line (NCI-H460). To achieve this, the luciferase signal, hPrx 6 mRNA expression, and superoxide dismutase (SOD) activity were measured in transfectants with a hPrx 6/Luc plasmid after treatment with four antioxidant extracts, including Korea white ginseng (KWG), Korea red ginseng (KRG), Liriope platyphylla (LP), and red Liriope platyphylla (RLP). First, the hPrx 6/Luc plasmid was successfully constructed with DNA fragments of human Prx 6 promoter, amplified by PCR using genomic DNA isolated from NCI-H460 cells, and cloned into the pTransLucent reporter vector. The orientation and sequencing of the hPrx 6/Luc plasmid were identified with restriction enzyme and automatic sequencing. A luciferase assay revealed significant enhancement of luciferase activity in the four treatment groups compared with a vehicle-treated group, although the ratio of the increase was different within each group. The KRG- and LP-treated groups showed higher activity than the KWG- and RLP-treated groups. Furthermore, the luciferase activity against RLP occurred roughly in a dose-dependent manner. However, the level of endogenous hPrx 6 mRNA did not change in any group treated with the four extracts. The SOD activity was in agreement with the luciferase activity. Therefore, these results indicate that the hPrx 6/Luc vector system may successfully express and respond to antioxidant compounds in NCI-H460 cells. The data also suggest that the Prx 6/Luc vector system may be effectively applied in screening the response of hPrx 6 to antioxidant compounds in transgenic mice.

• 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.

• Korean Journal of Organic Agriculture
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• v.25 no.1
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• pp.135-148
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• 2017

Response Surface Methodology (RSM), which is combining with Plackett-Burman design and Box-Behnken experimental design, was applied to optimize the ratios of the nutrient components for carotenoid production by Rhodobacter sphaeroides PS-24 in liquid state fermentation. Nine nutrient ingredients containing yeast extract, sodium acetate, NaCl, $K_2HPO_4$, $MgSO_4$, mono-sodium glutamate, $Na_2CO_3$, $NH_4Cl$ and $CaCl_2$ were finally selected for optimizing the medium composition based on their statistical significance and positive effects on carotenoid yield. Box-Behnken design was employed for further optimization of the selected nutrient components in order to increase carotenoid production. Based on the Box-Behnken assay data, the secondary order coefficient model was set up to investigate the relationship between the carotenoid productivity and nutrient ingredients. The important factors having influence on optimal medium constituents for carotenoid production by Rhodobacter sphaeroides PS-24 were determined as follows: yeast extract 1.23 g, sodium acetate 1 g, $NH_4Cl$ 1.75 g, NaCl 2.5 g, $K_2HPO_4$ 2 g, $MgSO_4$ 1.0 g, mono-sodium glutamate 7.5 g, $Na_2CO_3$ 3.71 g, $NH_4Cl$ 3.5g, $CaCl_2$ 0.01 g, per liter. Maximum carotenoid yield of 18.11 mg/L was measured by confirmatory experiment in liquid culture using 500 L fermenter.