• Title/Summary/Keyword: Functional characterization

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Isolation and Characterization of a Marine Bacterium Producing Thermotolerant Agarase (내열성 한천분해효소를 생산하는 해양세균의 분리 및 특성)

  • Park Ceun-Tae;Lee Dong-Ceun;Kim Nam Young;Lee Eo-Jin;Jung Jong-Ceun;Lee Jae-Hwa;Heo Moon-Soo;Lee Jung-Hyun;Kim Sang-Jin;Lee Sang-Hyeon
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.884-888
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    • 2005
  • An agar-degrading bacterium was isolated from north-eastern sea of Jeju island and cultured in marine agar 2216 media. Biochemical and morphologicl characteristics and 165 rRNA gene revealed that isolated strain was member of Agarivorans genus, and named Agarivorans sp. JA-1. Agarase was produced as growth-related and expressed regardless of agar presence. Optimal pH was 8 at 50 mM Clycine-NaOH buffer, and activity was maximum at $40^{\circ}C$E Enzymatic activity was maintained over $80\%$ at $60^{\circ}C$t and $70\%$ at $80^{\circ}C$ which is thermotolerant. Hence isolated novel Agarivorans sp. JA-1 strain and its beta-agarase could be used for the production of functional oligosaccharide from agar in solution state.

Characterization of Genes Related to the Cell Size Growth and CCN Family According to the Early Folliculogenesis in the Mouse (쥐의 초기 난포 발달에 관여하는 Cell Size Growth 및 CCN Family 유전자에 관한 연구)

  • Kim, Kyeoung-Hwa;Park, Chang-Eun;Yoon, Se-Jin;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.3
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    • pp.269-277
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    • 2005
  • Objectives: Previously, we sought to compile a list of genes expressed during early folliculogenesis by using cDNA microarray to investigate follicular gene expression and changes during primordialprimary follicle transition and development of secondary follicles (Yoon et al., 2005). Among those genes, a group of genes related to the cell size growth was characterized during the ovarian development in the present study. Methods: We determined ovarian expression pattern of six genes related to the cell size growth (cyr61, emp1, fhl1, socs2, wig1 and wisp1) and extended into CCN family (${\underline{c}}onnective$ tissue growth factor/${\underline{c}}ysteine$-rich 61/${\underline{n}}ephroblastoma$-overexpressed), ctgf, nov, wisp2, wisp3, including cyr61 and wisp1 genes. Expression of mRNA and protein according to the ovarian developmental stage was evaluated by in situ hybridization, and/or semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR), and immunohistochemistry, respectively. Results: Among 6 genes related to the cell size growth, cyr61 and wisp1 mRNA was detected only in oocytes in the postnatal day5 mouse ovaries. cyr61 mRNA expression was limited to the nucleolus of oocytes, while wisp1 was expressed in the cytoplasm and nucleolus of oocytes, except nucleus. cyr61 mRNA expression, however, was found in granulosa cells from secondary follicles. The rest 4 genes in the cell size growth group were detected in oocytes, granulosa and theca cells. Cyr61 and Wisp1 proteins were expressed in the oocyte cytoplasm from primordial follicle stage. Especially, Cyr61 protein was detected in pre-granulosa cells, Wisp1 protein was not. By using RT-PCR, we evaluated and decided that Cyr61 protein is produced by their own mRNA in pre-granulosa cells that was not detected by in situ hybridization. cyr61 and wisp1 genes are happen to be the CCN family members. The other members of CCN family were also studied, but their expression was detected in oocytes, granulose and theca cells. Conclusions: We firstly characterized the ovarian expression of genes related to the cell size growth and CCN family according to the early folliculogenesis. Cyr61 protein expression in the pre-granulosa cells is profound in meaning. Further functional analysis for cyr61 in early folliculogenesis is under investigation.

Identification and Functional Characterization of Two Noncoding RNAs Transcribed from Putative Active Enhancers in Hepatocellular Carcinoma

  • Lee, Ye-Eun;Lee, Jiyeon;Lee, Yong Sun;Jang, Jiyoung Joan;Woo, Hyeonju;Choi, Hae In;Chai, Young Gyu;Kim, Tae-Kyung;Kim, TaeSoo;Kim, Lark Kyun;Choi, Sun Shim
    • Molecules and Cells
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    • v.44 no.9
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    • pp.658-669
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    • 2021
  • Enhancers have been conventionally perceived as cis-acting elements that provide binding sites for trans-acting factors. However, recent studies have shown that enhancers are transcribed and that these transcripts, called enhancer RNAs (eRNAs), have a regulatory function. Here, we identified putative eRNAs by profiling and determining the overlap between noncoding RNA expression loci and eRNA-associated histone marks such as H3K27ac and H3K4me1 in hepatocellular carcinoma (HCC) cell lines. Of the 132 HCC-derived noncoding RNAs, 74 overlapped with the eRNA loci defined by the FANTOM consortium, and 65 were located in the proximal regions of genes differentially expressed between normal and tumor tissues in TCGA dataset. Interestingly, knockdown of two selected putative eRNAs, THUMPD3-AS1 and LINC01572, led to downregulation of their target mRNAs and to a reduction in the proliferation and migration of HCC cells. Additionally, the expression of these two noncoding RNAs and target mRNAs was elevated in tumor samples in the TCGA dataset, and high expression was associated with poor survival of patients. Collectively, our study suggests that noncoding RNAs such as THUMPD3-AS1 and LINC01572 (i.e., putative eRNAs) can promote the transcription of genes involved in cell proliferation and differentiation and that the dysregulation of these noncoding RNAs can cause cancers such as HCC.

Characterization of Miniimonas sp. S16 isolated from activated sludge (활성슬러지로부터 분리된 Miniimons sp. S16 세균의 특성)

  • Koh, Hyeon-Woo;Kim, Hongik;Park, Soo-Je
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.242-247
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    • 2019
  • Biological factors (e.g. microorganism activity) in wastewater treatment plant (WWTP) play essential roles for degradation and/or removal of organic matters. In this study, to understand the microbial functional roles in WWTP, we tried to isolate and characterize a bacterial strain from activated sludge sample. Strain S16 was isolated from the activated sludge of a municipal WWTP in Daejeon metropolitan city, the Republic of Korea. The cells were a Gram-stain-positive, non-motile, facultative anaerobe, and rod-shaped. Strain S16 grew at a temperature of $15{\sim}40^{\circ}C$ (optimum, $30^{\circ}C$), with 0~9.0% (w/v) NaCl (optimum, 1.0~2.0%), and at pH 5.5~9.0 (optimum, pH 7.0~7.5). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain S16 was most closely related to the unique species Miniimonas arenae NBRC $106267^T$ (99.79%, 16S rRNA gene sequence similarity) of the genus Miniimonas. The cell wall contained alanine, glutamic acid, serine, and ornithine. Although the isolation source of the type strain NBRC $106267^T$ which considered as a marine microorganism is sea sand, that of strain S16 is terrestrial environment. It might raise an ecological question for habitat transition. Therefore, comparative genome analysis will be valuable investigation for shedding light on their potential metabolic traits and genomic streamlining.

Preparation and characterization of Environmental Functional Nanofibers by electrospun nanofibers-Dry sorption material for indoor CO2 capture (정전방사를 통한 환경기능성 미세섬유 제조 및 특성분석 - 실내환경 CO2 포집용 건식흡착소재)

  • Kim, Eun Joo;Park, Kyung-Ryul
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.12
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    • pp.938-943
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    • 2018
  • Thin nano-sized fibres were prepared by an electrospinning method. The spinning appratus consisted of pump for polymer injection, nozzle and nozzle rotus, and an aluminum plate collected the polymer fibers. Its surface was chemically modified for selective improved adsorption of carbon monoxide at indoor level. The chemical activation enabled to form the fibres 250-350 nm in thickness with pore sizes distributed between 0.6 and 0.7 nm and an average specific surface area of $569m^2/g$. The adsorption capacities of pure (100%) and indoor (0.3%) $CO_2$, of which level frequently appears, at the ambient condition were improved from 1.08 and 0.013 to 2.2 and 0.144 mmol/g, respectively. It was found that the adsorption amount of $CO_2$ adsorbed by the chemically activated carbon nanofiber prepared through chemical activation would vary depending on the ratio of specific surface area and micropores. In particular, chemical interaction between adsorbent surface and gas molecules could enhance the selective capture of weak acidic $CO_2$.

Production of green tea jelly using theanine and its physiochemical characterization (녹차 theanine을 이용한 젤리 제조 및 품질특성 조사)

  • Kim, Seong Gyung;Jeong, Hana;Im, Ae Eun;Yang, Kwang-Yeol;Choi, Yong Soo;Nam, Seung-Hee
    • Korean Journal of Food Science and Technology
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    • v.53 no.5
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    • pp.553-560
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    • 2021
  • Theanine, the major amino acid and a sweet umami component of green tea, has anti-stress effects in humans. From green tea, theanine was extracted at 80℃ for 2 h using a low temperature, high pressure extractor, and caffeine was removed using an HP-20 column with 80% ethanol. Theanine extracts were applied to produce functional jelly using three kinds of gelling agents (I, II, and III) or various concentrations of theanine extracts (10-50%). Theanine jelly was characterized with respect to its physical properties, product stability, and physiological function. Gelling agent III (tamarind gum, xanthan gum, and locust bean gum=2:3:5, w/w/w) and S3 (35% theanine extracts) jelly exhibited the optimum textural properties with lower hardness and high springiness. Among theanine jellies, S3 exhibited optimum product stability, high 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging, and acetylcholinesterase inhibitory activity. These results indicate that the anine extracts could be used as a neuroprotective source in the food industry.

Immobilization of As and Pb in Contaminated Soil Using Bead Type Amendment Prepared by Iron NanoparticlesImpregnated Biochar (철 나노 입자가 담지된 바이오차 기반 비드 형태 안정화제를 이용한 비소 및 납 오염토양의 안정화)

  • Choi, Yu-Lim;Kim, Dong-Su;Kang, Tae-Jun;Yang, Jae-Kyu;Chang, Yoon-Young
    • Journal of Environmental Impact Assessment
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    • v.30 no.4
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    • pp.247-257
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    • 2021
  • In this work, Iron Nano-Particles Impregnated BioChar/bead (INPBC/bead) soil amendment was developed to increase biochar's reactivity to As in soil and preventing possible wind loss. Prior to preparation of INPBC/bead, INPBC was produced utilizing lignocellulosic biomass and Fe(III) solution in a hydrothermal method, followed by a calcination process. Then, the bead type amendment, INPBC/bead was produced by cross-linking reaction of alginate with INPBC. FT-IR, XRD, BET, and SEM-EDS analyses were utilized to characterize the as-synthesised materials. The particle size range of INPBC/bead was 1-4 mm, and different oxygen-containing functional groups and Fe3O4 crystalline phase were produced on the surface of INPBC/bead, according to the characterization results. The soil cultivation test was carried out in order to assess the stabilization performance of INPBC/bead utilizing As and Pb-contaminated soil obtained from an abandoned mining location in South Korea. After 4 weeks of culture, TCLP and SPLP extraction tests were performed to assess the stabilization efficacy of the amendment. The TCLP and SPLP findings revealed that raising the application ratio improved stabilizing efficiency. The As stabilization efficiency was determined to be 81.56 % based on SPLP test findings for a 5% in (w/w) INPBC/bead treatment, and the content of Pb in extracts was reduced to the limit of detection. According to the findings of this study, INPBC/bead that can maintain pH of origin soil and minimize wind loss might be a potential amendment for soil polluted with As and heavy metals.

Analysis of 16S rRNA gene sequencing data for the taxonomic characterization of the vaginal and the fecal microbial communities in Hanwoo

  • Choi, Soyoung;Cha, Jihye;Song, Minji;Son, JuHwan;Park, Mi-Rim;Lim, Yeong-jo;Kim, Tae-Hun;Lee, Kyung-Tai;Park, Woncheoul
    • Animal Bioscience
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    • v.35 no.11
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    • pp.1808-1816
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    • 2022
  • Objective: The study of Hanwoo (Korean native cattle) has mainly been focused on meat quality and productivity. Recently the field of microbiome research has increased dramatically. However, the information on the microbiome in Hanwoo is still insufficient, especially relationship between vagina and feces. Therefore, the purpose of this study is to examine the microbial community characteristics by analyzing the 16S rRNA sequencing data of Hanwoo vagina and feces, as well as to confirm the difference and correlation between vaginal and fecal microorganisms. As a result, the goal is to investigate if fecal microbiome can be used to predict vaginal microbiome. Methods: A total of 31 clinically healthy Hanwoo that delivered healthy calves more than once in Cheongju, South Korea were enrolled in this study. During the breeding season, we collected vaginal and fecal samples and sequenced the microbial 16S rRNA genes V3-V4 hypervariable regions from microbial DNA of samples. Results: The results revealed that the phylum-level microorganisms with the largest relative distribution were Firmicutes, Actinobacteria, Bacteroidetes, and Proteobacteria in the vagina, and Firmicutes, Bacteroidetes, and Spirochaetes in the feces, respectively. In the analysis of alpha, beta diversity, and effect size measurements (LefSe), the results showed significant differences between the vaginal and fecal samples. We also identified the function of these differentially abundant microorganisms by functional annotation analyses. But there is no significant correlation between vaginal and fecal microbiome. Conclusion: There is a significant difference between vaginal and fecal microbiome, but no significant correlation. Therefore, it is difficult to interrelate vaginal microbiome as fecal microbiome in Hanwoo. In a further study, it will be necessary to identify the genetic relationship of the entire microorganism between vagina and feces through the whole metagenome sequencing analysis and meta-transcriptome analysis to figure out their relationship.

Isolation of Simiduia sp. SH-2 and Characterization of Its β-Agarase (한천분해세균 Simiduia sp. SH-2 균주의 분리 및 β-agarase의 특성조사)

  • Lee, Dong-Geun;Kim, Geun-Dae;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.32 no.10
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    • pp.778-783
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    • 2022
  • This study isolated a new agarase-producing bacterium and characterized its agarase. A new agar-degrading strain was isolated from the seashore of Namhae in Gyeongnam province, Korea, and was purely cultured using the Marine Agar 2216 media. The isolated bacterium was identified as Simiduia sp. SH-2 after 16S rRNA gene sequencing. The crude agarase was obtained from the culture medium of the Simiduia sp. SH-2 strain, and the agar-degrading activity was measured. The highest level of activity of the Simiduia sp. SH-2-derived agar-degrading enzyme was 625 U/l. Agar degradation activity was most significant at 40℃ and pH 7.0. Compared to the activity at 40℃, the relative activity was 31% at 20℃ and 71% at 30℃. Compared to the activity at pH 7.0, the relative activity was 94% and 89% at pH 6.0 and pH 8.0, respectively. Residual activity was greater than 96% after exposure to 20℃ and 30℃ for 2 hr and more than 49% after exposure to 40℃ for 2 hr. Simiduia sp. SH-2 was identified as a strain producing β-agarase that creates neoagarooligosaccharides, such as neoagarotetraose and neoagarohexaose. Therefore, the Simiduia sp. SH-2 strain and its β-agarase are expected to be useful functional material producers in the food, cosmetic, and pharmaceutical industries.

Functional characterization of naturally-occurring constitutively activating/inactivating mutations in equine follicle-stimulating hormone receptor

  • Byambaragchaa, Munkhzaya;Ahn, Tae-Young;Choi, Seung-Hee;Kang, Myung-Hwa;Min, Kwan-Sik
    • Animal Bioscience
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    • v.35 no.3
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    • pp.399-409
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    • 2022
  • Objective: Follicle-stimulating hormone (FSH) is the central hormone involved in mammalian reproduction, maturation at puberty, and gamete production that mediates its function by control of follicle growth and function. The present study investigated the mutations involved in the regulation of FSH receptor (FSHR) activation. Methods: We analyzed seven naturally-occurring mutations that were previously reported in human FSHR (hFSHR), in the context of equine FSHR (eFSHR); these include one constitutively activation variant, one allelic variant, and five inactivating variants. These mutations were introduced into wild-type eFSHR (eFSHR-wt) sequence to generate mutants that were designated as eFSHR-D566G, -A306T, -A189V, -N191I, -R572C, -A574V, and -R633H. Mutants were transfected into PathHunter EA-parental CHO-K1 cells expressing β-arrestin. The biological function of mutants was analyzed by quantitating cAMP accumulation in cells incubated with increasing concentrations of FSH. Results: Cells expressing eFSHR-D566G exhibited an 8.6-fold increase in basal cAMP response, as compared to that in eFSHR-wt. The allelic variation mutant eFSHR-A306T was not found to affect the basal cAMP response or half maximal effective concentration (EC50) levels. On the other hand, eFSHR-D566G and eFSHR-A306T displayed a 1.5- and 1.4-fold increase in the maximal response, respectively. Signal transduction was found to be completely impaired in case of the inactivating mutants eFSHR-A189V, -R572C, and -A574V. When compared with eFSHR-wt, eFSHR-N191I displayed a 5.4-fold decrease in the EC50 levels (3,910 ng/mL) and a 2.3-fold decrease in the maximal response. In contrast, cells expressing eFSHR-R633H displayed in a similar manner to that of the cells expressing the eFSHR-wt on signal transduction and maximal response. Conclusion: The activating mutant eFSHR-D566G greatly enhanced the signal transduction in response to FSH, in the absence of agonist treatment. We suggest that the state of activation of the eFSHR can modulate its basal cAMP accumulation.