• Title/Summary/Keyword: KRIBB

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Identification of a Second Type of AHL-Lactonase from Rhodococcus sp. BH4, belonging to the α/β Hydrolase Superfamily

  • Ryu, Du-Hwan;Lee, Sang-Won;Mikolaityte, Viktorija;Kim, Yea-Won;Jeong, Haeyoung;Lee, Sang Jun;Lee, Chung-Hak;Lee, Jung-Kee
    • Journal of Microbiology and Biotechnology
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    • v.30 no.6
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    • pp.937-945
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    • 2020
  • N-acyl-homoserine lactone (AHL)-mediated quorum sensing (QS) plays a major role in development of biofilms, which contribute to rise in infections and biofouling in water-related industries. Interference in QS, called quorum quenching (QQ), has recieved a lot of attention in recent years. Rhodococcus spp. are known to have prominent quorum quenching activity and in previous reports it was suggested that this genus possesses multiple QQ enzymes, but only one gene, qsdA, which encodes an AHL-lactonase belonging to phosphotriesterase family, has been identified. Therefore, we conducted a whole genome sequencing and analysis of Rhodococcus sp. BH4 isolated from a wastewater treatment plant. The sequencing revealed another gene encoding a QQ enzyme (named jydB) that exhibited a high AHL degrading activity. This QQ enzyme had a 46% amino acid sequence similarity with the AHL-lactonase (AidH) of Ochrobactrum sp. T63. HPLC analysis and AHL restoration experiments by acidification revealed that the jydB gene encodes an AHL-lactonase which shares the known characteristics of the α/β hydrolase family. Purified recombinant JydB demonstrated a high hydrolytic activity against various AHLs. Kinetic analysis of JydB revealed a high catalytic efficiency (kcat/KM) against C4-HSL and 3-oxo-C6 HSL, ranging from 1.88 x 106 to 1.45 x 106 M-1 s-1, with distinctly low KM values (0.16-0.24 mM). This study affirms that the AHL degrading activity and biofilm inhibition ability of Rhodococcus sp. BH4 may be due to the presence of multiple quorum quenching enzymes, including two types of AHL-lactonases, in addition to AHL-acylase and oxidoreductase, for which the genes have yet to be described.

Effects of Protox Herbicide Tolerance Rice Cultivation on Microbial Community in Paddy Soil (Protox 제초제저항성 벼 재배가 토양미생물 군집에 미치는 영향)

  • Oh, Sung-Dug;Ahn, Byung-Ohg;Kim, Min-Kyeong;Sohn, Soo-In;Ryu, Tae-Hun;Cho, Hyun-Suk;Kim, Chang-Gi;Back, Kyoung-Whan;Lee, Kijong
    • Korean Journal of Environmental Agriculture
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    • v.32 no.2
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    • pp.95-101
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    • 2013
  • BACKGROUND: Rice (Oryza sativa) is the most important staple food of over half the world's population. This study was conducted to evaluate the possible impact of transgenic rice cultivation on the soil microbial community. METHODS AND RESULTS: Microorganisms were isolated from the rhizosphere of GM and non-GM rice cultivation soils. Microbial community was identified based on the culture-dependent and molecular biology methods. The total numbers of bacteria, fungi, and actinomycete in the rhizosphere soils cultivated with GM and non-GM rice were similar to each other, and there was no significant difference between GM and non-GM rice. Dominant bacterial phyla in the rhizosphere soils cultivated with GM and non-GM rice were Actinobacteria, Firmicutes, and Proteobacteria. The microbial communities in GM and non-GM rice cultivated soils were characterized using the denaturing gradient gel electrophoresis (DGGE). The DGGE profiles showed similar patterns, but didn't show significant difference to each other. DNAs were isolated from soils cultivating GM and non-GM rice and analyzed for persistence of inserted gene in the soil by using PCR. The PCR analysis revealed that there were no amplified protox gene in soil DNA. CONCLUSION(S): These data suggest that transgenic rice does not have a significant impact on soil microbial communities, although continued research may be necessary.

The Effect of Multi-lamellar Emulsion (MLE) on Skin Barrier Function: Can an Improve Permeability Barrier Provide a Solution for Itching due to Skin Barrier Malfunction\ulcorner

  • Youm, Jong-kyung;Kim, Yang-hee;Park, Byeong-deog;Jeong, Se-kyoo;Park, Eung-ho;Ahn, Sung-ku;Lee, Seung-hun
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.765-779
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    • 2003
  • Physiological lipid mixtures comprised of cholesterol, ceramide and free fatty acid better maintain epidermal homeostasis and have been recently used for dermatoses induced by skin barrier damage, for example for atopic dermatitis and xerotic skin. Itching and dry atopic dermatitis of the skin may be related to altered skin barrier function. In a previous study, the use of multi-lamellar emulsion (MLE), which is a lipid mixtures containing cholesterol, pseudoceramide and free fatty acid, has been shown to accelerate the recovery of the epidermal permeability barrier. In this study, we assessed the efficacy of MLE compared with a currently used anti-itch moisturizer (AIM), the active ingredients of which are menthol and camphor, on barrier recovery after barrier disruption. To clarify the effect of MLE and AIM after acute barrier perturbation, we measured the relation between transepidermal water loss (TEWL) and the barrier recovery rate at 3, 6, 24, and 48 hours after tape stripping hairless mice and then observed changes in the stratum corneum (SC), including the intercellular lipid structure and secretion of lamellar bodies, by electron microscopy. MLE treated skin recover skin barrier function more rapidly, and AIM treated skin delayed barrier repair. Morphological changes in the epidermis, of MLE treated skin revealed well-conserved lipid multi-lamellar structures at 24 h after tape stripping, whereas AIM treated skin showed altered lamellar bilayers within the SC interstices at 48 h. In addition, MLE treated skin showed an increase in the number of LBs and in their secretions and a decrease in the number of SC layers versus AIM treated skin. These results suggest that MLE may accelerate the production of an epidermal permeability barrier in hairless mice by increasing the number and secretion of LB and improve the dryness and itch associated with an altered epidermal permeability barrier.

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Production of High Quality Pork with Feed Additives of Lacquer Tree Extracts and Development of Meat Processing Technique V. Lymphocyte Distribution and Morphological Change in Small Intestine of Pigs after Fed with Fermented Lacquer Tree Extracts (옻 추출물을 이용한 돈육의 생산과 가공기술의 개발 V. 옻 추출물 급여 후 돼지 림프구의 분포와 소장 점막 내부의 형태 변화)

  • Kim, Sang-Hyun;Seuong, Nak-Ju;Park, Joong-Choon;Choi, Chung-Kuk;Song, Young-Min;Cho, Kyu-Woan;Cha, Hye-Jin;Kim, Yong-Hwan
    • Journal of Veterinary Clinics
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    • v.25 no.4
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    • pp.268-273
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    • 2008
  • For evaluation of immune stimulation effect of lacquer tree extracts, lymphocytes were counted by labeling of FITC-conjugated monoclonal antibody in the pheripheral blood of pigs that fed with a fodder supplemented by lacquer tree extracts. Populations of MHC-II+, CD4+, and CD8+T lymphocytes were increased more than 2% level after 1 week feed supply of the lacquer tree extracts. The increase of those T cells reached at maximum level after 2 weeks in the tested group. B lympyocytes with surface IgM were increased 5% after 1 week feed supply of the lacquer tree extracts, and their numbers reached maximum after 2 weeks in the tested group. For the assessment of cytotoxicity of the lacquer tree extracts, morphological changes were examined on the epithelial cells of small intestine from pigs fed with a fodder supplemented by 0.1 % lacquer tree extracts for 6 weeks (the tested group). Thin-sectioned tissue of small intestine was fixed with glutaraldehyde, then coated with gold particles, and the specimen was examined under scanning electron microscope. The villi on the mucus membrane of jejunum and ileum from the tested pigswere enlarged on the tip and were linked each other.

Anomalous somatic embryos formation and plant regeneration from the cultures of immature embryos of Camellia japonica L. (동백나무 미숙배 배양으로부터 비정상 체세포배 형성과 식물체 재생)

  • Choi, Jong-Hye;Kwon, Suk-Yoon;Choi, Pil-Son
    • Journal of Plant Biotechnology
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    • v.38 no.4
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    • pp.258-262
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    • 2011
  • Embryogenic callus was induced from the cultures of immature embryos of Camellia japonica L. on Murashige & Skoog's (MS) solid medium supplemented with 1 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D), and then the embryogenic callus was proliferated on same medium for 4 weeks over. The embryogenic callus was sub-cultured on MS basal medium without 2,4-D to produce coyledonary stage of somatic embryo. The frequency (%) of somatic embryogenesis was 25.1%, and the majority of somatic embryos formed had a abnormal morphology with cupshaped cotyledon (48.3%), one cotyledon (12.6%), three cotyledons (9.4%), four cotyledons (1.9%), whereas was only normal morphology with two cotyledon (27.5%). When the somatic embryos with normal or abnormal cotyledons transfer to MS basal medium or $\frac{1}{2}$ MS medium with/or without plant growth regulators ($GA_3$, IBA) for regeneration, the frequency (%) of two-cotyledon embryos regenerated into plantlets was higher 11.1% than one cotyledon (0.0~8.3 %), three cotyledons (0.0~5.8%), four cotyledons (0.0%), cup-shaped (0.3~4.2%). These results demonstrated that the anomalous cotyledons of somatic embryos could caused to decrease the rate of plant regeneration.

Single Nucleotide Polymorphisms linked to the SlMYB12 Gene that Controls Fruit Peel Color in Domesticated Tomatoes (Solanum lycopersicum L.)

  • Kim, Bichsaem;Kim, Nahui;Kang, Jumsoon;Choi, Youngwhan;Sim, Sung-Chur;Min, Sung Ran;Park, Younghoon
    • Horticultural Science & Technology
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    • v.33 no.4
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    • pp.566-574
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    • 2015
  • Yellow or transparent fruit peel color is caused by the accumulation or lack of naringenin chalcone (NG, C) in fruit peel and determines the red or pink appearance of tomato fruit, respectively. NGC biosynthesis is regulated by the SlMYB12 gene of the Y locus on chromosome 1, and DNA markers derived from SlMYB12 would be useful for marker-assisted selection (MAS) of tomato fruit color. To develop a gene-based marker, 4.9 kb of the SlMYB12 gene including a potential promoter region was sequenced from the red-fruited (YY) line 'FCR' and pink-fruited (yy) line 'FCP'. Sequence alignment of these SlMYB12 alleles revealed no sequence variations between 'FCR' and 'FCP'. To identify SlMYB12-linked single nucleotide polymorphisms (SNPs), 'FCR' and 'FCP' were genotyped using a SolCAP Tomato SNP array and CAPS markers (CAPS-456, 531, 13762, and 38123) were developed from the four SNPs (solcap_snp_sl_456, 531, 13762, and 38123) most closely flanking the SlMYB12. These CAPS markers were mapped using $F_2$ plants derived from 'FCR' ${\times}$ 'FCP'. The map positions of the fruit peel color locus (Y) were CAPS-13762 (0 cM) - 456 (11.09 cM) - Y (15.71 cM) - 38123 (17.82 cM) - 531 (30.86 cM), and the DNA sequence of SlMYB12 was physically anchored in the middle of CAPS-456 and CAPS-38123, indicating that fruit peel color in domesticated tomato is controlled by SlMYB12. A total of 64 SolCAP tomato germplasms were evaluated for their fruit peel color and SNPs located between solcap_snp_sl_456 and 38123. Seven SNPs that were detected in this interval were highly conserved for pink-fruited accessions and specific to transparent fruit peel traits, as depicted by a phenetic tree of 64 accessions based on the seven SNPs.

Stable Transformation via Callus Formation and Rhizogenesis from the Cultures of Hypocotyl Explant of Chinese Cabbage (배추의 배축절편으로부터 캘러스와 뿌리 발생을 통한 안정적 형질전환)

  • Cho, Mi-Ae;Kim, Choon-Ae;Min, Sung-Ran;Ko, Suck-Min;Liu, Jang-Ryol;Choi, Pil-Son
    • Journal of Plant Biotechnology
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    • v.34 no.2
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    • pp.139-144
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    • 2007
  • Hypocotyl explants of Chinese cabbage (cvs. "Jeong Sang") produced transgenic calli on callus induction medium (MS salt, B5 vitamin, 5 mg/L acetosyringone, 1 mg/L 2,4-D, 3% sucrose, 400 mg/L cefotaxime, 100 mg/L paromomycin, pH 5.8) after cocultivation with strains of Agrobacterium tumefaciens (EHA101, LBA4404, GV3101) harboring the pPTN290 containing paromomycin-resistance gene as a selectable marker, and then they transferred to root induction medium (1/2MS salt, MS vitamins, 2% sucrose, 100 mg/L paromomycin, 100 mg/L cefotaxime, pH 5.8) and shoot induction medium (MS salt, B5 vitamin, 4 mg/L $AgNO_3$, 4 mg/L 6-benzyladenine, 3 mg/L alpha-naphthaleneacetic acid, 100 mg/L paromomycin, 100 mg/L cefotaxime, 3% sucrose, pH 5.8) in order. There was a significant difference in the frequency of transgenic calli depending on Agrobacterium strains. In particular, the highest frequency (6.1%) of transgenic calli was obtained from the hypocotyls cocultivated with EHA101 strains. Also, the frequency (%) of transgenic root and plants from each transgenic callus clone were obtained with 60.7% and 38.2% in EHA101, with 8.3% and 0% in LBA4404, with 20.5% and 85.7% in GV3101 strains, respectively. They were grown to maturity in a greenhouse and normally produced $T_2$ seeds. GUS histochemical assay for progeny ($T_2$) revealed that the transgenes was expressed in the plant genome, and progeny analysis from 7 independent transgenic events demonstrated that the transformants transmitted the transgene as a single or multiple functional locus.

Analysis of the Effect of Carbon Dioxide Reduction by Changing from Signalized Intersection to Roundabout using Tier 3 Method (Tier 3 방법을 이용한 회전교차로 도입에 따른 $CO_2$ 감축효과)

  • Lee, Jung-Beom;Lee, Seung-Hoon
    • The Journal of The Korea Institute of Intelligent Transport Systems
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    • v.10 no.5
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    • pp.105-112
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    • 2011
  • Delay reduction of vehicles at the intersection is highly dependent on the signal operation method. Improper traffic operation causes the violation of the traffic regulations and increasing traffic congestion. Delay because of congestion has contributed to the increase in carbon dioxide in the atmosphere. The focus of this paper is to measure the amount of carbon dioxide when the intersection is changed to roundabout. Even though, Intergovernmental Panel on Climate Change(IPCC) recommends Tier 1 method to measure the amount of greenhouse gas from vehicles, this paper used Tier 3 method because we could use the data of average running distance per each vehicle model. Two signalized intersections were selected as the study area and the delay reductions of roundabout operation were estimated by VISSIM microscopic simulation tool. The control delay for boksu intersection reduced from 28.6 seconds to 4.4 seconds and the KRIBB intersection sharply reduced from 156.4 seconds to 23.6 seconds. In addition, carbon dioxide for two intersections reduced to 646.5 ton/year if the intersection is changed to roundabout. Future research tasks include testing the experiment for networks, as well as for various intersection types.

Regulatory roles of ganglioside GQ1b in neuronal cell differentiation of mouse embryonic stem cells

  • Kwak, Dong-Hoon;Jin, Jung-Woo;Ryu, Jae-Sung;Ko, Kinram;Lee, So-Dam;Lee, Jeong-Woong;Kim, Ji-Su;Jung, Kyu-Yong;Ko, Ki-Sung;Ma, Jin-Yeul;Hwang, Kyung-A;Chang, Kyu-Tae;Choo, Young-Kug
    • BMB Reports
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    • v.44 no.12
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    • pp.799-804
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    • 2011
  • Gangliosides play an important role in neuronal differentiation processes. The regulation of ganglioside levels is related to the induction of neuronal cell differentiation. In this study, the ST8Sia5 gene was transfected into mESCs and then differentiated into neuronal cells. Interestingly, ST8Sia5 gene transfected mESCs expressed GQ1b by HPTLC and immunofluorescence analysis. To investigate the effects of GQ1b over-expression in neurogenesis, neuronal cells were differentiated from GQ1b expressing mESCs in the presence of retinoic acid. In GQ1b expressing mESCs, increased EBs formation was observed. After 4 days, EBs were co-localized with GQ1b and nestin, and GFAP. Moreover, GQ1b co-localized with MAP-2 expressing cells in GQ1b expressing mESCs in 7-day-old EBs. Furthermore, GQ1b expressing mESCs increased the ERK1/2 MAP kinase pathway. These results suggest that the ST8Sia5 gene increases ganglioside GQ1b and improves neuronal differentiation via the ERK1/2 MAP kinase pathway.

Neuroprotective effects of Rg3-enriched Korean Red Ginseng on alcohol-induced apoptosis in PC12 Cells (PC12 세포에서 알코올 유발성 세포 사멸에 대한 Rg3 풍부 고려 홍삼의 신경세포 보호 효과)

  • Choi, Na-Eun;Ryu, Jin-Hyeob;Lee, Dong-Ha;Cho, Hyun-Jeong
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.12
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    • pp.521-528
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    • 2017
  • Excessive alcohol consumption is one of the leading causes of many neurological diseases, such as dementia and Alzheimer's disease, and many efforts are under way to solve them. Red ginseng is known to enhance neuronal survival, inhibit apoptosis, and promote nerve regeneration of nerve cells. This study examined whether Rg3-enriched Korean red ginseng extract (KRG) inhibits the apoptosis of PC12 cells caused by alcohol-induced neurotoxicity and how KRG regulates several factors related to the caspase mediated pathway. In this way, the cell survival rate and apoptosis rate of PC12 cells were measured using an EZ-Cytox cell viability assay kit and flow cytometry, respectively. The expression of the apoptosis-related proteins (Bcl-2, Bid, Bax and caspase-3) were analyzed by western blotting, and the significance of the measured results was confirmed using the ANOVA method. As a result, KRG increased the expression of Bcl-2; inhibited the expression of Bid, Bax, and caspase-3; and inhibited the apoptosis of alcohol-induced PC12 cells. These results mean that the KRG-induced increase in Bcl-2 expression and down-regulation of Bid and Bax expression down-regulate caspase-3 expression, which in turn inhibits the mitochondrial apoptotic pathways. This study suggests that KRG is worth developing as a neuroprotective agent candidate.