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Discovery of a Novel Cellobiose Dehydrogenase from Cellulomonas palmilytica EW123 and Its Sugar Acids Production

  • Ake-kavitch Siriatcharanon;Sawannee Sutheeworapong;Sirilak Baramee;Rattiya Waeonukul;Patthra Pason;Akihiko Kosugi;Ayaka Uke;Khanok Ratanakhanokchai;Chakrit Tachaapaikoon
    • Journal of Microbiology and Biotechnology
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    • v.34 no.2
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    • pp.457-466
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    • 2024
  • Cellobiose dehydrogenases (CDHs) are a group of enzymes belonging to the hemoflavoenzyme group, which are mostly found in fungi. They play an important role in the production of acid sugar. In this research, CDH annotated from the actinobacterium Cellulomonas palmilytica EW123 (CpCDH) was cloned and characterized. The CpCDH exhibited a domain architecture resembling class-I CDH found in Basidiomycota. The cytochrome c and flavin-containing dehydrogenase domains in CpCDH showed an extra-long evolutionary distance compared to fungal CDH. The amino acid sequence of CpCDH revealed conservative catalytic amino acids and a distinct flavin adenine dinucleotide region specific to CDH, setting it apart from closely related sequences. The physicochemical properties of CpCDH displayed optimal pH conditions similar to those of CDHs but differed in terms of optimal temperature. The CpCDH displayed excellent enzymatic activity at low temperatures (below 30℃), unlike other CDHs. Moreover, CpCDH showed the highest substrate specificity for disaccharides such as cellobiose and lactose, which contain a glucose molecule at the non-reducing end. The catalytic efficiency of CpCDH for cellobiose and lactose were 2.05 × 105 and 9.06 × 104 (M-1 s-1), respectively. The result from the Fourier-transform infrared spectroscopy (FT-IR) spectra confirmed the presence of cellobionic and lactobionic acids as the oxidative products of CpCDH. This study establishes CpCDH as a novel and attractive bacterial CDH, representing the first report of its kind in the Cellulomonas genus.

Community Analysis and Bological Water Quality Evaluation of Benthic Macroinvertebrate in Wangpi-cheon Watershed (왕피천 유역의 저서성 대형무척추동물 군집분석 및 생물학적 수질평가)

  • Park, Young-Jun;Jeon, Yong-Lak;Kim, Ki-Dong;Yoon, Hee-Nam;Nam, Sang-Ho
    • Korean Journal of Environment and Ecology
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    • v.27 no.3
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    • pp.327-343
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    • 2013
  • The aim of this study is to perform community analysis and biological assessment of water quality using benthic macroinvertebrate from Wangpi-cheon watershed which is defined as conservation areas of ecosystem and landscape by ministry of environment in Korea. Field survey of the study area was carried out 2 times from June to September in 2012. As a result of the field survey, total 155 species of benthic macroinvertebrates in 74 families, 15 orders, 7 classes and 5 phyla were collected. The findings of community analysis using the classified species and individuals showed relatively low DI(Dominant Index) value of 0.22 and very high value of average H'(Diversity index) as 4.24. And the analyzed results of SI(Similarity Index) according to habitat types using functional feeding groups showed higher values of 94.51% and 93.19% respectively to tributary and main stream after the designation of conservation areas of ecosystem and landscape. These results infer that various species and lots of individual are widely distributed at Wangpi-cheon watershed and stream ecosystem of the study area is healthy and well maintained after the designation of conservation areas. And also, the calculated EPT value was 62.9% as high enough to explain the cleanness of Wangpi-cheon watershed. We evaluated environmental condition and biological water quality by using ESB(Ecological Score of Benthic macroinvertebrate community) and KSI(Korean Saprobic Index). The average evaluated ESB shows very high value of 208.2, therefore Wangpi-cheon watershed is designated as 'First priority protection waters' area and the value of KSI is 0.32 which meets the saprobic water quality standard as 'First class'.

Quantitative Expression Analysis of Functional Genes in Four Dog Breeds (개의 네 품종에서 기능 유전자들에 대한 정량적 발현 분석)

  • Gim, Jeong-An;Kim, Sang-Hoon;Lee, Hee-Eun;Jeong, Hoim;Nam, Gyu-Hwi;Kim, Min Kyu;Huh, Jae-Won;Choi, Bong-Hwan;Kim, Heui-Soo
    • Journal of Life Science
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    • v.25 no.8
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    • pp.861-869
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    • 2015
  • One of the domesticated species; the dog has been selectively bred for various aims by human. The dog has many breeds, which are artificially selected for specific behaviors and morphologies. Dogs contribute their life to human as working dogs for guide, rescue, detection or etc. Working dogs requires good personality, such as gentleness, robustness and patience for performing their special duty. Many studies have concentrated on finding genetic marker for selecting the high-quality working dog. In this study, we confirmed quantitative expression patterns of eight genes (ABAT; 4-Aminobutyrate Aminotransferase, PLCB1; Phospholipase C, Beta 1, SLC10A4; Solute Carrier Family 10, Member 4, WNT1; Wingless-Type MMTV Integration Site Family, Member 1, BARX2; BarH-Like Homeobox 2, NEUROD6; Neuronal Differentiation 6, SEPT9; Septin 9 and TBR1; T-Box, Brain, 1) among brains tissues from four dog breeds (Beagle, Sapsaree, Shepherd and Jindo), because these genes were expressed and have functions in brain mostly. Specially, BARX2, SEPT9, SLC10A4, TBR1 and WNT1 genes were highly expressed in Beagle and Jindo, and Sapsaree and German Shepherd were vice versa. The biological significance of total genes was estimated by database for annotation, visualization and integrated discovery (DAVID) to determine a different gene ontology (GO) class. In these analyses, we suppose to these eight genes could provide influential information for brain development, and intelligence of organisms. Taken together, these results could provide clues to discover biomarker related to functional traits in brain, and beneficial for selecting superior working dogs.

Vegetation Structure and Population Dynamics of Berchemia racemosa Habitats (청사조(Berchemia racemosa) 자생지의 식생구조 및 개체군 동태 분석)

  • Beon, Mu-Sup;Kim, Young-Ha
    • Korean Journal of Environment and Ecology
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    • v.22 no.6
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    • pp.679-690
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    • 2008
  • The objectives of this study are to investigate and analyze the vegetation structure and population dynamics of Berchemia racemosa habitats in the Weolmyung park in Gunsan city, and base on that to seek the ecological habitat conservation plan for the Berchemia racemosa. In results, the Berchemia racemosa habitats are located at $81{\sim}93$ meters above the sea level, in steep seaside slope of a mountain. The soil texture are silt loam mainly and soil pH were $4.1{\sim}5$. The vascular plants in the Berchemia racemosa habitats has been analyzed as 61 taxa; 33 families, 51 genera, 54 species, 6 varieties, and 1 forms. Berchemia racemosa as a Specific plant species by floral region was the class V. Berchemia racemosa habitats were classified into 7 vegetation communities of Quercus serrata community(A1), Alnus firm a community(A2), Platycarya strobilacea community(A3), Robinia pseudoacacia community(A4) and 3 Pinus densiflora communities(B1, B2, B3). The importance value of Berchemia racemosa were 30%(A1), 15%(A2), 27%(A3), 65%(A4), 18%(B1), 45%(B2) and 35%(B3) on shrubs layer and 12, 27, 20, 18, 11, 18, 21 % on herb layer. The constant companion species with Berchemia racemosa were Stephanandra incisa and Ligustrum obtusifolium. Total 103 populations appear in the 7 Berchemia racemosa habitats. Their spatial distribution pattern were clumped for the most part. The average height was 133cm, the root color diameter was 4.4cm and the ramification branch number was 9.4. From the results of this study, it is suggested the continued monitoring and the active protection measures for the Berchemia racemosa habitats.

Selection of (Ac/Ds) insertion mutant lines by abiotic stress and analysis of gene expression pattern of rice (Oryza sativar L.) (비생물학적 스트레스 관련 벼 Ac/Ds 삽입 변이체의 선발 및 유전자 발현 분석)

  • Jung, Yu-Jin;Park, Seul-Ah;Ahn, Byung-Ohg;Yun, Doh-Won;Ji, Hyeon-So;Lee, Gang-Sup;Park, Young-Whan;Suh, Seok-Cheol;Baek, Hyung-Jin;Lee, Myung-Chul
    • Journal of Plant Biotechnology
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    • v.35 no.4
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    • pp.307-316
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    • 2008
  • Transposon-mediated insertional mutagenesis is one of powerful strategy for assessing functions of genes in higher plants. In this report, we have selected highly susceptible and tolerance plant by screening about high salt (3% NaCl) and cold stresses ($4^{\circ}C$) from F2 seeds of 30,000 Ac/Ds insertional mutagenesis lines in rice (Oryza sativa L. cv. Dongjin). In order to identify the gene tagging, insertion of Ds element was analyzed by Southern blot and these results revealed that 19 lines were matched genotype of selected lines with phenotype from the first selected 212 lines, and 13 lines have one copy of Ds elements. The Franking Sequence Tags (FSTs) of selected mutant lines showed high similarities with the following known function genes: signal transduction and regulation of gene expression (transpoter, protease family protein and apical meristem family protein), osmotic stress response (heat shock protein, O-methyltransferase, glyceraldehyde-3-phosphate dehydrogenase and drought stress induce protein), vesicle trafficking (SYP 5 family protein) and senescence associated protein. The expression pattern of 19 genes were analyzed using RT-PCR under the abiotic stresses of 9 class; 250mM NaCl, osmotic, drought, 3% $H_2O_2$, $100{\mu}M$ ABA, $100{\mu}M$ IAA, 0.1 ppm 2,4-D, $4^{\circ}C$ cold and $38^{\circ}C$ high temperature. Isolated knock-out genes showed the positive response about 250 mM NaCl, drought, $H_2O_2$, PEG, IAA, 2,4-D, ABA treatment and low ($4^{\circ}C$) and high temperature ($38^{\circ}C$). The results from this study indicate that function of selected knock-out genes could be useful in improving of tolerance to abiotic stresses as an important transcriptional activators in rice.

THE STUDY ON THE EFFECTS OF THE INSULIN-LIKE GROWTH FACTOR-I ON THE BIOLOGICAL ACTIVITY OF THE HUMAN PERIODONTAL LIGAMENT CELLS (Insulin-like growth factor-I 이 치주인대세포의 생물학적 활성도에 미치는 영향에 대한 연구)

  • Kim, Seong-Jin;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.24 no.2
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    • pp.219-237
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    • 1994
  • The ultimate goal of clinical periodontal therapy is to achieve regeneration of a healthy connective tissue reattachment. Conventional therapy including scaling, root planing, gingival curettage, gingivectomy and flap procedures of various types results primarily in repair rather than regeneration of the periodontium. In order for periodontal regeneration to occur, progenitor periodontal ligament cells must migrate to the denuded root surface, attach to it, proliferate and mature into an organized and functional fibrous attachment apparatus. Polypeptide growth factors belong to a class of potent biologic mediators which regulate cell differentiation, proliferation, migration and metabolism. Insulin-like growth factor-I (IGF- I ) of these factors appear to have an important role in periodontal wound healing and bone formation. The purpose of this study is to evaluate the effects of IGF- I on the periodontal ligament cells to use as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were obtained from periodontal tissue explants culture of the first premolar tooth extracted for the orthodontic treatment. Cells were cultured in Dulbecco's modified Eagle medium(DMEM) with 10% fetal bovine serum. Fourth to seventh passage cells were plated in 24 well tissue culture plates and medium changed to serum-free medium prior to addition of growth factors. Cell proliferation was measured by the incorporation of $[^3H]-thymidine$ into DNA, Protein synthesis was determined by measurement of $[^3H]-proline$ incorporation into collagenase-digestible protein(CDP) and noncollagenous protein(NCP) according to the method of Peterkofsky and Diegelmann (1971), And alkaline phosphatase activity was measured as one parameter of osteoblastic differentiation. The results were as follows : The DNA synthetic activity was increased in a dose-dependent manner with IGF- I except for 0.1ng/ml concentration of IGF- I At the concentration of 10, 100ng/ml, IGF- I significantly increased the DNA synthetic activity(P<0.05) The total protein, collagen and noncollagen synthesis was increased in a dose-dependent manner with IGF- I except for 0.1ng/ml concentration of IGF- I. At the concentration of 1, 10, 100ng/ml, IGF- I significantly increased the total protein, collagen and noncollagen synthesis activity(P<0.95, P<0.001). The % of collagen was not effected according to the concentration of IGF- I. The alkaline phosphatase activity was increased in a dose-, time-dependent manner with IGF- I (10, 100ng/ml). In conclusions, the present study shows that IGF- I has a potentiality to enhance the DNA synthesis of periodontal ligament cells with including the increase of the total protein and collagen synthetic activity. The use of IGF- I to mediate biological stimulation of periodontal ligament cells shows promise for future therapeutic applications.

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Effect of Soil Water Potential on Pysico-Chemical Properties of Soil and Cucumber(Cucumis sativus L.) Growth (토양(土壤) 수분(水分)포텐셜이 오이(Cucumis sativus L.)생육(生育)과 토양(土壤) 이화학적(理化學的 ) 특성(特性)에 미치는 영향(影響))

  • Bum, In-Sook;Kim, Yong-Woong;Kim, Kwang-Sik;Kim, Kil-Yong;Sohn, Bo-Kyoon;Kim, Hyun-Woo
    • Korean Journal of Soil Science and Fertilizer
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    • v.32 no.2
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    • pp.171-181
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    • 1999
  • A field experiments was carried out to investigate the physicochemical properties of soil and cucumber growth in vinyl house when irrigation point was made at 0.2, 1/3, 0.5 and 1.0 bar. The obtained results was summarized as follow: The taxonomic class of the soil used was loam and each content of the required water was 4.4, 7.3, 9.6 and 13.4 mm per each irrigation time at 0.2, 1/3, 0.5 and 1.0 bar treatments in spring culture, respectively. At 0.2 bar and 1.0 bar treatments, interval of irrigation was 2.3 and 14.8 day, the times of irrigation was 37 and 6, and total irrigation volume was 163.5 and 80.3 mm, respectively. After cucumber culture, pH, EC concentration and exchangeable K content of soil at 0.2 bar treatment was distributed near to the level of improvement target while EC, available $P_2O_5$ and exchangeable base content in other treatments were higher compared to improvement target. At 1.0 bar treatment, ratios of the solid and liquid phase were 44.9 and 27.1%, respectively, and bulk density was $1.26g\;cm^{-3}$ which was the highest among the treatments. At 0.2 bar treatment, the ratio of the solid and liquid phase was 41.7 and 22.8%, respectively, and bulk density was $1.09g\;cm^{-3}$ which was the lowest. The root length and radius at 0.2 bar treatment were best, while those at 1/3 bar were worst. At 0.2 bar treatment, the total yield was 7,269 kg and the weight of good products was 5,677 kg which was the highest among treatments. At 0.33 bar treatment, the yield was the lowest with the high ratio of deformity.

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Representative Physical and Chemical Properties of Korean Soils by the Results from Detailed Soil Survey (우리나라 토양(土壤)의 대표적(代表的)인 물리화학적(物理化學的) 특성(特性) (정밀토양조사결과(精密土壤調査結果)를 중심(中心)으로))

  • Hur, Bong-Koo;Jo, In-Sang;Min, Kyeong-Beom;Um, Ki-Tae
    • Korean Journal of Soil Science and Fertilizer
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    • v.17 no.4
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    • pp.330-336
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    • 1984
  • These studies were aimed to find out the representative values of physico-chemical properties in Korean soils and to serve the basic informations to improve the soil properties for increasing the soil productivity. By the results of detailed soil survey, 14 physico-chemical properties (soil texture, moisture characteristics, organic matter content etc) were collected from 315 series of soils except Cheju soils, computerized the simple mean and the distributed area by weighted mean values with grouping of land use, drainage class and soil type. The results were as follows; 1. Simple mean values within the efficient soil depth in whole country soils were clay 20.0%, organic matter 2.03% and CEC 10.3me/100g. But weighted mean values with land area belonged to the same soil series were clay 18.0%, OM 1.85% and CEC 8.6me/100g. 2. Mean values of paddy soils weighted by area were clay 19.7%, OM 2.0% and CEC 9.1me/100g but those of upland soils were 17.9%, 1.8% and 8.1me/100g and forest soils were 16.7%, 1.8% and 8.6me/100g respectively. 3. Weighted mean values of moderately well soils with covered area were clay 18.9%, organic matter 1.7%, CEC 8.4me/100g but those of imperfectly drained soils were 21.3%, 2.2% and 9.5me/100g, and those of poorly drained soils were 15.1%, 1.8% and 9.9me/100g respectivcly. 4. Simple mean and area weighted mean values of clay content, pH, organic matter contents, cation exchange capacity and base saturation were calculated by the types of paddy and upland.

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Winter Algal Bloom and Spatial Characteristics of Water Quality in the Lower Taewha River, Ulsan, Korea (태화강 하류에서 겨울철 조류 발생과 수질의 공간적 특성)

  • Sohn, Eun Rak;Park, Jung Im;Lee, Bora;Lee, Jin Woo;Kim, Jongseol
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.30-37
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    • 2013
  • This study was carried out to assess the spatial and tidal effects on the water quality in the lower reaches of Taewha River, Ulsan, Korea and to understand the environmental factors affecting winter algal bloom in the river. From May, 2010 to January, 2011, water samples were collected at five locations (New Samho Bridge, Old Samho Bridge, Mungjung Stream, Taewha Bridge, and Mungchon Bridge) along the river at high and low tides of spring tide. We measured environmental parameters including salinity, dissolved oxygen (DO), biological oxygen demand (BOD), chemical oxygen demand (COD), chlorophyll a (Chl a) and various nutrient concentrations. Salinity increased towards the downstream direction. Average values of Chl a concentrations ranged $10-26mg/m^3$ at high tide and $11-53mg/m^3$ at low tide depending on sampling locations. It was noteworthy that there were strong increases in Chl a concentrations during the November 21 to December 22 sampling period especially at the Taewha Bridge. At the location, Chl a concentrations were measured as $138-296mg/m^3$ for the period; Rhodomonas lacustris of class Cryptophyceae was the dominant algal species. Chl a concentrations at the Taewha Bridge were positively correlated with such parameters as salinity, BOD, DO, COD, pH, and T-N, and negatively correlated with temperature and $NO_3{^-}$-N. On the other hand, at the Mungchon Bridge the highest concentration of Chl a was $55mg/m^3$ on August 25, and Chl a concentrations were positively correlated with $NH_3$-N, T-N, $PO_4{^{3-}}$-P, T-P, and heterotrophic plate counts. The results suggested that water quality in the lower Taewha River fluctuated a lot with the sampling locations and the patterns of algal blooms were different between Taewha Bridge and Mungchon Bridge sampling locations.

Analysis of Global Gene Expression Profile of Human Adipose Tissue Derived Mesenchymal Stem Cell Cultured with Cancer Cells (암세포주와 공동 배양된 인간 지방 조직 유래 중간엽 줄기 세포의 유전자 발현 분석)

  • Kim, Jong-Myung;Yu, Ji-Min;Bae, Yong-Chan;Jung, Jin-Sup
    • Journal of Life Science
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    • v.21 no.5
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    • pp.631-646
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    • 2011
  • Mesenchymal stem cells (MSC) are multipotent and can be isolated from diverse human tissues including bone marrow, fat, placenta, dental pulp, synovium, tonsil, and the thymus. They function as regulators of tissue homeostasis. Because of their various advantages such as plasticity, easy isolation and manipulation, chemotaxis to cancer, and immune regulatory function, MSCs have been considered to be a potent cell source for regenerative medicine, cancer treatment and other cell based therapy such as GVHD. However, relating to its supportive feature for surrounding cell and tissue, it has been frequently reported that MSCs accelerate tumor growth by modulating cancer microenvironment through promoting angiogenesis, secreting growth factors, and suppressing anti-tumorigenic immune reaction. Thus, clinical application of MSCs has been limited. To understand the underlying mechanism which modulates MSCs to function as tumor supportive cells, we co-cultured human adipose tissue derived mesenchymal stem cells (ASC) with cancer cell lines H460 and U87MG. Then, expression data of ASCs co-cultured with cancer cells and cultured alone were obtained via microarray. Comparative expression analysis was carried out using DAVID (Database for Annotation, Visualization and Integrated Discovery) and PANTHER (Protein ANalysis THrough Evolutionary Relationships) in divers aspects including biological process, molecular function, cellular component, protein class, disease, tissue expression, and signal pathway. We found that cancer cells alter the expression profile of MSCs to cancer associated fibroblast like cells by modulating its energy metabolism, stemness, cell structure components, and paracrine effect in a variety of levels. These findings will improve the clinical efficacy and safety of MSCs based cell therapy.