• The Korean Journal of Malacology
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• v.28 no.4
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• pp.377-384
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• 2012

The importance of biological resources has been gradually increasing, and mollusks have been utilized as main fishery resources in terrestrial ecosystems. But little is known about genomic and transcriptional analysis in mollusks. This is the first report on the transcriptomic profile of Meretrix lusoria. In this study, we constructed cDNA library and determined 542 of distinct EST sequences composed of 284 singletons and 95 contigs. At first, we identified 180 of EST sequences that have significant hits on protein sequences of the exclusive Mollusks database through BLASTX program and 343 of EST sequences that have significant hits on NCBI NR database. We also found that 211 of putative sequences through local BLAST (blastx, E < e-10) search against KOG database were classified into 16 functional categories. Some kinds of immune response related genes encoding allograft inflammatory factor 1 (AIF-1), B-cell translocation gene 1 (BTG1), C-type lectin A, thioester-containing protein and 26S proteasome regulatory complex were identified. To determine phylogenetic relationship, we identified partial sequences of four genes (COX1, COX2, 12S rRNA and NADH dehydrogenase) that significantly matched with the mitochondrial genomes of 3 species-Ml (Meretrix lusoria), Mp (Meretrix petechialis) and Mm (Meretrix meretrix). As a result, we found that there was a little bit of a difference between sequences of Korean isolates and other known isolates. This study will be useful to develop breeding technology and might also be helpful to establish a classification system.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.8
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• pp.1131-1140
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• 2014

This research focused on the effects of different doses of Bacillus subtilis KN-42 on the growth performance, diarrhea incidence, faecal bacterial flora, and the relative number of Lactobacillus and Escherichia coli in faeces of weaned piglets to determine whether the strain can serve as a candidate antimicrobial growth promoter. A total of 360 piglets (initial body weight $7.14{\pm}0.63$ kg) weaned at $26{\pm}2$ days of age were randomly allotted to 5 treatment groups (4 pens per treatment with 18 pigs per pen) for a 28-day trial. Dietary treatments were basal diet without any antimicrobial (negative control; NC), basal diet supplemented with 120 mg/kg feed of neomycin sulfate (positive control; PC) and basal diet supplemented with $2{\times}10^9$ (L), $4{\times}10^9$ (M) and $20{\times}10^9$ (H) CFU/kg feed of B. subtilis KN-42. During the overall period, average daily gain and feed efficiency of piglets were higher in groups PC, M, and H than those in group NC (p<0.05), and all probiotics and antibiotics groups had a lower diarrhea index than group NC (p<0.05). The 16S rDNA gene-based methods were used to analyze faecal bacterial flora on day 28 of experiment. The result of denaturing gradient gel electrophoresis analysis showed that supplementation of B. subtilis KN-42 to the diet changed the bacterial communities, with a higher bacterial diversity and band number in group M than in the other four groups. Real-time polymerase chain reaction analysis showed that the relative number of Lactobacillus were higher in groups PC and H than in group NC (p<0.05), and the supplemented B. subtilis KN-42 to the diet also reduced the relative number of E. coli (p<0.05). These results suggest that dietary addition of B. subtilis KN-42 can improve the growth performance and gastrointestinal health of piglets.

• The Korean Journal of Mycology
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• v.41 no.4
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• pp.218-224
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• 2013

Of 1,100 yeast strains which were isolated from various Korean fermented foods, screened for phosphate solubilization, five strains showed the ability to solubilize tricalcium phosphate. The 26S rDNA domain D1-D2 sequence analysis revealed the identification of strain Y393 and Y524 as Pichia anomala (99.8 and 100% identity, respectively), Y669 as Pichia farinosa (100% identity), Y901 as Candida versatilis (100% identity), and Y1101 as Pichia subpelliculosa (100% identity). All the phosphate solubilizing strains showed mesophilic characteristics. The temperature range for growth of 4 strains was $20{\sim}35^{\circ}C$ and P. farinosa Y669 was able to grow up to $45^{\circ}C$. The strain C. versatilis Y907 was able to grow at pH range of 5.0~6.0 and showed halophilic characteristics with tolerance to 15% of NaCl concentration. The Phosphate solubilizing yeast strains were survived well in bed soil for 8 weeks which were maintained densities of $10^7{\sim}10^8$ cfu/g. The highest phosphate solubilizing activity was observed in P. subpelliculosa Y1101. It solubilized 697.2 ug/mL of phosphorus from tricalcium phosphate with decrease in pH from 6.8 to 4.37 after 11 days of inoculation.

• The Plant Pathology Journal
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• v.25 no.1
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• pp.86-90
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• 2009

In November 2008, typical powdery mildew symptoms were observed on leaves of Arabidopsis thaliana ecotype Col-0 plants in a growth room under controlled laboratory conditions at Korea University, Seoul. The disease was characterized by the appearance of white powder-like fungal growth on the surface of infected leaves. As the disease progressed, infected leaves exhibited chlorotic or necrotic brown lesions, and leaf distortion and senescence. Conidiophores of the causal fungus were hyaline, unbranched, 3-4 celled, cylindrical, and $80-115{\times}6-9{\mu}m$ in size. Singly produced conidia (pseudoidium type) were hyaline, oblong to cylindrical or oval in shape, and $26-55{\times}15-20{\mu}m$ in size with a length/width ratio of average 3, angular/rectangular wrinkling of outer wall and no distinct fibrosin bodies. Appressoria on the hyphae were multi-lobed. These structures are typical of the powdery mildew Oidium subgenus Pseudoidium, anamorph of the genus Erysiphe. The measurements of the fungal structures coincided with those of Erysiphe cruciferarum. The phylogenetic analysis using ITS rDNA sequences revealed that the causal fungus Erysiphe sp. KUS-F23994 is identical to E. cruciferarum. The isolated fungus incited powdery mildew symptoms on the inoculated Arabidopsis leaves, which proved Koch's postulates. Taken all data together, we first report the occurrence of powdery mildew disease of A. thaliana caused by Erysiphe cruciferarum in Korea.

• The Korean Journal of Mycology
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• v.45 no.3
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• pp.234-240
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• 2017

The goal of this study was to isolate wild yeasts from reed fields in the Seocheon-gun county in Chungcheongnam-do, South Korea. Molecular analysis of the D1/D2 domain of the 26S rDNA of the yeasts isolated from soil samples was performed using Basic Local Alignment Search Tool (BLAST). In total, 20 yeast strains from 11 species were isolated from 20 different soil samples. Candida species were observed to be the most common yeasts. Of these 20 strains, Candida subhashii (6 strains), Candida tropicalis (3 strains), Candida boleticola MS008, and Lachancea thermotolerans (2 strains) were identified as alcohol-fermenting yeasts. Further, Bullera japonica YJ10-1, Candida subhashii J7-1, Kluyveromyces yarrowii YJ11-1, and Ustilago shanxiensis Y10-1 were newly recorded yeast strains in Korea, and therefore, their microbiological characteristics were investigated further. All of these unrecorded yeast strains had oval cells; only Candida subhashii J7-1 formed ascospores and pseudomycelia. Kluyveromyces yarrowii YJ11-1 grew in vitamin-free medium, and all of these strains, except Candida subhashii J7-1, grew in 5% NaCl-containing YPD broth.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.11
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• pp.1555-1561
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• 2016

Shank skin color of Korean native chicken (KNC) shows large color variations. It varies from white, yellow, green, bluish or grey to black, whilst in the majority of European breeds the shanks are typically yellow-colored. Three shank skin color-related traits (i.e., lightness [$L^*$], redness [$a^*$], and yellowness [$b^*$]) were measured by a spectrophotometer in 585 progeny from 68 nuclear families in the KNC resource population. We performed genome scan linkage analysis to identify loci that affect quantitatively measured shank skin color traits in KNC. All these birds were genotyped with 167 DNA markers located throughout the 26 autosomes. The SOLAR program was used to conduct multipoint variance-component quantitative trait locus (QTL) analyses. We detected a major QTL that affects $b^*$ value (logarithm of odds [LOD] = 47.5, $p=1.60{\times}10^{-49}$) on GGA24 (GGA for Gallus gallus). At the same location, we also detected a QTL that influences $a^*$ value (LOD = 14.2, $p=6.14{\times}10^{-16}$). Additionally, beta-carotene dioxygenase 2 (BCDO2), the obvious positional candidate gene under the linkage peaks on GGA24, was investigated by the two association tests: i.e., measured genotype association (MGA) and quantitative transmission disequilibrium test (QTDT). Significant associations were detected between BCDO2 g.9367 A>C and $a^*$ ($P_{MGA}=1.69{\times}10^{-28}$; $P_{QTDT}=2.40{\times}10^{-25}$). The strongest associations were between BCDO2 g.9367 A>C and $b^*$ ($P_{MGA}=3.56{\times}10^{-66}$; $P_{QTDT}=1.68{\times}10^{-65}$). However, linkage analyses conditional on the single nucleotide polymorphism indicated that other functional variants should exist. Taken together, we demonstrate for the first time the linkage and association between the BCDO2 locus on GGA24 and quantitatively measured shank skin color traits in KNC.

• Food Science and Preservation
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• v.24 no.7
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• pp.1043-1051
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• 2017

Wild yeasts were isolated from domestic non-sterilized Makgeolli and their fermentation characteristics were analyzed to select the best fermentation seed culture. A total of 65 yeast strains isolated yeasts from non-sterilized Makgeolli and Nuruk. In order to select fermentable strains, hydrogen sulfide, $CO_2$ production ability, alcohol tolerance and aroma component production ability were analyzed. To screen the aromatic strains of isolates, media containing cerulenin, 5,5,5-trifluor-DL-leucine (TFL) and API ZYM kit were used. There were 36 strains resistance to cerulenin and all strains produced esterase and demonstrated tolerance against TFL. Hydrogen sulfide, which could degrade the quality of the fermented beverage, was not produced in 34 yeast. The correlation between alcohol tolerance of yeast and carbon dioxide production was analyzed by principal component analysis. YM22, YM31, YM32 and YM37 produced a total of 0.14-0.18 g/72 h of $CO_2$ indicating high fermentability. Alcohol tolerance was measured by alcohol concentration. YM32, YM37 yeast had 20% alcohol tolerance. As a result, alcohol and flavor characteristics of wild yeast isolated from non-sterilized Makgeolli were analyzed and it was confirmed that yeast was suitable for the production of alcohol.

• Archives of Pharmacal Research
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• v.21 no.5
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• pp.581-590
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• 1998

We developed a novel water-soluble camptothecin analobue, CKD602, and evaluated the inhibition of topoisomerase I and the antitumor activities against mammalian tumor cells and human tumor xenografts. CKD602 was a nanomolar inhibitor of the topoisomerase I enzyme in the cleavable complex assay. CKD602 was found to be 3 times and slightly more potent than topotecan and camptothecin as inhibitors of topoisomerase, respecitively. In tumor cell cytotoxicity, CKD602 was more potent than topotecan in 14 out of 26 human cancer cell lines tested, while it was comparable to camptothecin. CKD602 was tested for the in vivo antitumor activity against the human tumor xenograft models. CKD602 was able to imduce regression of established HT-29, WIDR and CX-1 colon tumors, LX-1 lung tumor, MX-1 breast tumor and SKOV-3 ovarian tumor as much as 80, 94, 76, 67, 87% and 88%, respectively, with comparable body weight changes to those of topotecan. Also the therapeutic margin (R/Emax: maximum tolerance dose/$ED-{58}$) of CKD602 was significantly higher than that of topotecan by 4 times. Efficacy was determined at the maximal tolerated dose levels using schedule dependent i.p. administration in mice bearing L1210 leukemia. On a Q4dx4 (every 4 day for 4 doses) schedule, the maximum tolerated dose (MTD) was 25 mg/kg per administration, which caused great weight loss and lethality in <5% tumor bearing mouse. this schedule brought significant increase in life span (ILS), 212%, with 33% of long-term survivals. The ex vivo antitumor activity of CKD602 was compared with that of topotecan and the mean antitumor index (ATI) values recorded for CKD602 were significantly higher than that noted for topotecan. From these results, CKD602 warrants further clinical investigations as a potent inhibitor of topoisomerase I.

• Microbiology and Biotechnology Letters
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• v.26 no.2
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• pp.122-129
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• 1998

It were reported that antifungal mechanism of Enterobacter cloacae is a volatile ammonia that produced by the strain in soil, and the production of ammonia is related to the bacterial urease activity. A powerful bacterium SH14 against soil-borne pathogen Fusarium solani, which cause root rot of many important crops, was selected from a ginseng pathogen suppressive soil. The strain SH14 was identified as Bacillus subtilis by cultural, biochemical, morphological method, and $API^{circledR}$ test. From several in vitro tests, the antifungal substance that is produced from B. subtilis SH14 was revealed as heat-stable and low-molecular weight antibiotic substance. In order to construct the multifunctional biocontrol agent, the urease gene of Bacillus pasteurii which can produce pathogenes-suppressive ammonia transferred into antifungal bacterium. First, a partial BamH I digestion fragment of plasmid pBU11 containing the alkalophilic B. pasteurii l1859 urease gene was inserted into the BamH I site of pEB203 and expressed in Escherichia coli JM109. The recombinant plasmid was designated as pGU366. The plasmid pGU366 containing urease gene was introduced into the B. subtilis SH14 with PEG-induced protoplast transformation (PIP) method. The urease gene was very stably expressed in the transformant of B. subtilis SH14. Also, the optimal conditions for transformation were established and the highest transformation frequency was obtained by treatment of lysozyme for 90 min, and then addition of 1.5 ${mu}g$/ml DNA and 40% PEG4000. From the in vitro antifungal test against F. solani, antifungal activity of B. subtilis SH14(pGu366) containing urease gene was much higher than that of the host strain. Genetical development of B. subtilis SH14 by transfer of urease gene can be responsible for enhanced biocontrol efficacy with its antibiotic action.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2004.05a
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• pp.89-119
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• 2004

This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.