• 제목/요약/키워드: Taxonomic characterization

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Alaska 툰드라 토양의 깊이 및 해동 영향에 따른 미생물 군집과 토양 유기 탄소 분해 특성 (Characterization of microbial communities and soil organic carbon degradation associated with the depth and thawing effects on tundra soil in Alaska)

  • 박하주;김덕규;박현;이방용;이유경
    • 미생물학회지
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    • 제52권3호
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    • pp.365-374
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    • 2016
  • 고위도에서의 온도 상승은 $0.6^{\circ}C$/10 년으로, 이는 토양 유기 탄소에 대한 미생물의 분해 활성 증가를 유도한다. 게다가, 분해된 토양 유기 탄소는 이산화탄소 또는 메탄 같은 온실가스로 전환, 방출되어 기후 변화를 가속화시킨다. 따라서, 토양 유기 탄소 분해와 관련된 미생물의 다양성 및 기능 이해를 위한 토양 해동 모델 연구가 필요하다. 이러한 연구를 위하여 Alaska Council의 두 깊이의 토양(SPF와 PF라 각각 명명한 30-40와 50-60 cm 깊이의 토양)을 $0^{\circ}C$에서 108일 동안 배양하였다. 환경 모사 실험 동안 pyrosequencing을 수행하였고, metagenome을 분석하여 총 111,804개의 미생물 sequence를 얻었다. 이 중, 574-1,128개의 세균 operational taxonomic unit (OTU)과 30-57개의 고세균 OTU를 확인하였다. 토양 배양에 따라 두 토양 모두에서 Crenarchaeota phyla의 상대적 분포가 증가하였으며, Actinobacteria와 Firmicutes phyla의 분포가 SPF와 PF에서 각각 크게 증가하였다. 추출한 토양 유기 탄소에 대한 무게 측정 및 gel permeation chromatography를 통해, 환경 모사 실험이 진행되는 동안 토양 유기 탄소의 주요 구성 성분인 부식산(humic acids)이 중합화(humification)되는 것을 확인하였다. 결론적으로, 냉대 툰드라 동토의 해동은 Crenarchaeota, Actinobacteria 및 Firmicutes phyla의 증가를 야기시키며, 미생물에 의한 토양 유기 탄소 분해 및 이용을 야기시키는 것으로 예측된다.

Studies on Constituents of Higher Fungi of Korea (LXXI) -Application of Enzymes to Taxonomy of Ganoderma Species-

  • Kim, Byong-Kak;Kim, Jin-Sook;Choi, Kyun-Gae;Kim, Ha-Won;Choi, Eung-Chil
    • 생약학회지
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    • 제24권2호
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    • pp.116-123
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    • 1993
  • The genus Ganoderma is typical wood-rotting fungi and its fruiting body has been used as an important herb in oriental medicine. Recent research discovered antitumor components from Ganoderma lncidum. Various Ganoderma species are being cultivated in Korea. However, taxonomic system of the genus Ganoderma has been based mainly on the macromorphology of fruiting bodies and the ultrastructural characteristics of basidiospores. Since there are similar characteristics in Ganoderma mycelia grown on the same artificial media, it is suggested that the compatibility of the fungi by di-mon mating be used as an aid to determine the identity of species in addition to the conventional characterization. In this study, we examined physiological and genetical properties such as growth temperature, pH, compatibility and enzyme or protein patterns of laccase, esterase and cellular proteins of G. lucidum RZ, G. tsugae and Ganoderma species cultivated in Korea by electrophoresis for characterization of the isolates. We found that compatibility test and isozyme patterns of laccase and esterase of the mycelia could be used for the differentiation of the isolates. These results showed that Ganoderma species cultivated in Korea is genetically similar to G. lucidum but physiologically closer to G. tsugae than to G. lucidum.

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방선균으로부터 Cholesterol Oxidase의 생산 및 특성 (Production and Characterization of Cholesterol Oxidase from Streptomyces sp. No.4)

  • 김현수;고희선
    • KSBB Journal
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    • 제14권2호
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    • pp.174-180
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    • 1999
  • Cholesterol oxidase(EC.1.1.3.6)는 cholesterol을 산화 또는 이성화시키는 효소로, 혈중 cholesterol이 측정 등의 임상진단용 시약에 이용되고 있으며, 여려종류의 미생물에서 분리, 연구되어 왔다. 현재에는 농업 및 식품 등에도 응용이 기대되고 있는 매우 유용한 효소이다. 본 실힘은 토양에서 분리한 방선균으로부터 cho1esterol oxidase생산능이 우수한 균주를 선발하여 속의 동정, 효소의 생산조건 및 효소학적 특성을 조사하였다 . 본 분리균주는 배양학적, 형태학적 생리학적 특성 및 현미경 검정을 통하여 Streptomyces sp으로 동정하였다. 본 효소의 생산조건을 조사한 결괴 탄소원으로 1% soluble starch, 질소원으로 2% corn steep liquor가 효소생산에 우수하였으며, 이들 탄소원, 질소원이의 0.1% $NaNO_3$, 0.1% $KH_2PO_4$, 0.05% $MgSO_4$가 함유된 배지를 생산 최적 배지로 사용하였다. 본 효소의 특성을 검토한 결과, 최적온도와 최적pH는 각각 $37^{\circ}C$, pH 6.5~7.5이었으며. 효소와 안정성은 온도 $30-40^{\circ}C$에사 80% 이상 활성이 유지되었으며, pH 6.0~9.0의 범위까지 안정한 것으로 나타났다. 본 효소의 등전점은 multichambered electrofocusing unit를 사용하여 측정한 결과 pH 6.0-6.5인 것으로 추정되었다.

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Morphological and Molecular Characterization of Thamnocalamus falconeri Hook f. ex. Munro

  • Tiwari, Chandrakant;Bakshi, Meena;Nautiyal, Subhash
    • Journal of Forest and Environmental Science
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    • 제31권3호
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    • pp.214-224
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    • 2015
  • The economy of India and so also of many Asian countries depends on bamboos and their uses are not only in domestic items but also in rural housing and raw materials to several industries and germplasm characterization is an important link between the conservation and utilization of plant genetic resources. Classical taxonomic studies of the bamboos are based on floral morphology and growth habit, which can cause problems in identification due to erratic flowering coupled with different biotic agencies and environmental factors. Identification and genetic relationships among accessions of Thamnocalamus falconeri were investigated using morphology and random amplified polymorphic DNAs (RAPD) technique. Analysis started by using 51 vegetative characters and forty two 10-mer primers that allowed us to distinguish different genotypes hailing from different eco- zones of Garhwal Himalayas (India). The selected primers (12) were used for identification and for establishing a profiling system to estimate genetic diversity. A total of 79.33% polymorphism was estimated by using 12 selected primers. The genetic similar analysis was conducted based on binary digits i.e. presence (1) or absence (0) of bands, which revealed a wide range of variability among the species whereas genetic relatedness was quite high based on vegetative characters. Cluster analysis clearly showed two major clusters for both of the markers viz. morphology and RAPD belonging to 10 accessions of T. falconeri. Two major clusters were further divided into minor clusters. Cluster based on RAPD marker showed grouping of accessions of closed locality whereas analogy was reported for vegetative traits. The RAPD technique has the potential for use in species identification and genetic relationships studies of bamboo for breeding program.

Isolation, Production, and Characterization of Protease from Bacillus subtilis IB No. 11

  • Lee, Min-Hyang;Lee, Kang-Moon;Choi, Yong-Jin;Baek, Yeon-Soo
    • Journal of Animal Science and Technology
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    • 제51권6호
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    • pp.527-536
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    • 2009
  • A potent protein degrading bacterium was isolated from soil samples of different environments. Polyphasic taxonomic studies and phylogenetic 16S rRNA sequence analyses led to identify the isolate IB No. 11 as a strain of Bacillus subtilis. The isolated strain was recognized to produce protease constitutively, and the maximum production (1.64 units/ml) was attained in a shake flask culture when the isolate was grown at $40^{\circ}C$, for 32 h in basal medium supplemented with starch (0.25%) and gelatin (1.25%) as sole carbon and nitrogen source, respectively. The optimum pH and temperature for the protease activity were determined to be pH 7.0 and $50^{\circ}C$, respectively. $Ca^{2+}$ and $Mn^{2+}$ enhanced remarkably the protease activity but neither showed positive effect on the protease's thermal stability. In addition, it was observed that the protease was fairly stable in the pH range of 6.5-8.0 and at temperatures below $50^{\circ}C$, and it could be a good candidate for an animal feed additive. The inhibition profile of the protease by various inhibitors indicated that the enzyme is a member of serine-proteases. A combination of UV irradiation and NTG mutagenesis allowed to develop a protease hyper-producing mutant strain coded as IB No. 11-4. This mutant strain produced approximately 3.23-fold higher protease activity (6.74 units/mg) than the parent strain IB No. 11 when grown at $40^{\circ}C$ for 32h in the production medium. The protease production profile of the selected mutants was also confirmed by the zymography analysis.

Phylogenetic Diversity of Acidophilic Sporoactinobacteria Isolated from Various Soils

  • Cho, Sung-Heun;Han, Ji-Hye;Seong, Chi-Nam;Kim, Seung-Bum
    • Journal of Microbiology
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    • 제44권6호
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    • pp.600-606
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    • 2006
  • Spore forming actinobacteria (sporoactinobacteria) isolated from soils with an acidic pH in Pinus thunbergii forests and coal mine waste were subjected to taxonomic characterization. For the isolation of acidophilic actinobacteria, acidified starch casein agar (pH adjusted to 4-5) was used. The numbers of actinobacteria growing in acidic media were between $3.2{\times}10^4$ and $8.0{\times}10^6$ CFU/g soil. Forty three acidophilic actinobacterial strains were isolated and their 16S rDNA sequences were determined. The isolates were divided into eight distinctive phylogenetic clusters within the variation encompassed by the family Streptomycetaceae. Four clusters among them were assigned to the genus Streptacidiphilus, whereas the remaining four were assigned to Streptomyces. The clusters belonging to either Streptomyces or Streptacidiphilus did not form a monophyletic clade. The growth pH profiles indicated that the representative isolates grew best between pH 5 and 6. It is evident from this study that acidity has played a critical role in the differentiation of the family Streptomycetaceae, and also that different mechanisms might have resulted in the evolution of two groups, Streptacidiphilus (strict acidophiles) and neutrotolerant acidophilic Streptomyces. The effect of geographic separation was clearly seen among the Streptacidiphilus isolates, which may be a key factor in speciation of the genus.

Application of Molecular Biology to Rumen Microbes -Review-

  • Kobayashi, Y.;Onodera, R.
    • Asian-Australasian Journal of Animal Sciences
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    • 제12권1호
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    • pp.77-83
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    • 1999
  • Molecular biological techniques that recently developed, have made it possible to realize some of new attempts in the research field of rumen microbiology. Those are 1) cloning of genes from rumen microorganisms mainly in E. coli, 2) transformation of rumen bacteria and 3) ecological analysis with nonculturing methods. Most of the cloned genes are for polysaccharidase enzymes such as endoglucanase, xylanase, amylase, chitinase and others, and the cloning rendered gene structural analyses by sequencing and also characterization of the translated products through easier purification. Electrotransformation of Butyrivibrio fibrisolvens and Prevotella ruminicola have been made toward the direction for obtaining more fibrolytic, acid-tolerant, depoisoning or essential amino acids-producing rumen bacterium. These primarily required stable and efficient gene transfer systems. Some vectors, constructed from native plasmids of rumen bacteria, are now available for successful gene introduction and expression in those rumen bacterial species. Probing and PCR-based methodologies have also been developed for detecting specific bacterial species and even strains. These are much due to accumulation of rRNA gene sequences of rumen microbes in databases. Although optimized analytical conditions are essential to reliable and reproducible estimation of the targeted microbes, the methods permit long term storage of frozen samples, providing us ease in analytical work as compared with a traditional method based on culturing. Moreover, the methods seem to be promissing for obtaining taxonomic and evolutionary information on all the rumen microbes, whether they are culturable or not.

용담호 조류군집의 시공간적 분포와 조류발생 요인분석 (Characterization of Algal Community of Yongdam Reservoir and Identification of Ecological Factors Inducing the Changes in Community Composition)

  • 김현수;정일환
    • 한국지하수토양환경학회지:지하수토양환경
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    • 제20권7호
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    • pp.121-134
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    • 2015
  • Spatial and temporal changes in algal population in Yongdam reservoir and ecological factors that induced the changes in the size and composition of algal population were investigated by monthly sampling at ten locations in the reservoir. Nutritional state of the reservoir was identified to be phosphorus-limited with nitrogen to phosphorus (N : P) ratio much greater than 17 in most samples. Algal population was dominated by three taxonomic groups, diatoms, chlorophytes and cyanobacteria. Although explosive algal growth was not observed in the summer, algal population showed transition with time of the dominant algal type from diatoms in the winter to cyanobacteria in the summer. Chlorophyta was not the dominant group in the reservoir although they maintained relatively stable number of cells in the reservoir and showed increase in population from March to May. The application of statistical methods revealed that the factors inducing changes in cell number of each group were water temperature for diatoms and cyanobacteria and phosphorus concentration for chlorophyte. Fluctuation of cyanobacterial population was mainly observed near the inlet of tributaries while diatoms showed higher variation inside the reservoir.

Molecular Characterization of Nippostrongylus brasiliensis (Nematoda: Heligmosomatidae) from Mus musculus in India

  • Chaudhary, Anshu;Goswami, Urvashi;Singh, Hridaya Shanker
    • Parasites, Hosts and Diseases
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    • 제54권6호
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    • pp.743-750
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    • 2016
  • Mus musculus (Rodentia: Muridae) has generally been infected with a rodent hookworm Nippostrongylus brasiliensis. In this report, we present morphological and molecular identification of N. brasiliensis by light and scanning electron microscopy and PCR amplification of mitochondrial cytochrome c oxidase subunit 1 (cox1) gene and the protein sequences encoded by cox1 gene, respectively. Despite the use of N. brasiliensis in many biochemistry studies from India, their taxonomic identification was not fully understood, especially at the species level, and no molecular data is available in GenBank from India. Sequence analysis of cox1 gene in this study revealed that the present specimen showed close identity with the same species available in GenBank, confirming that the species is N. brasiliensis. This study represents the first record of molecular identification of N. brasiliensis from India and the protein structure to better understand the comparative phylogenetic characteristics.

Computational Approaches for Structural and Functional Genomics

  • Brenner, Steven-E.
    • 한국생물정보학회:학술대회논문집
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    • 한국생물정보시스템생물학회 2000년도 International Symposium on Bioinformatics
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    • pp.17-20
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    • 2000
  • Structural genomics aims to provide a good experimental structure or computational model of every tractable protein in a complete genome. Underlying this goal is the immense value of protein structure, especially in permitting recognition of distant evolutionary relationships for proteins whose sequence analysis has failed to find any significant homolog. A considerable fraction of the genes in all sequenced genomes have no known function, and structure determination provides a direct means of revealing homology that may be used to infer their putative molecular function. The solved structures will be similarly useful for elucidating the biochemical or biophysical role of proteins that have been previously ascribed only phenotypic functions. More generally, knowledge of an increasingly complete repertoire of protein structures will aid structure prediction methods, improve understanding of protein structure, and ultimately lend insight into molecular interactions and pathways. We use computational methods to select families whose structures cannot be predicted and which are likely to be amenable to experimental characterization. Methods to be employed included modern sequence analysis and clustering algorithms. A critical component is consultation of the presage database for structural genomics, which records the community's experimental work underway and computational predictions. The protein families are ranked according to several criteria including taxonomic diversity and known functional information. Individual proteins, often homologs from hyperthermophiles, are selected from these families as targets for structure determination. The solved structures are examined for structural similarity to other proteins of known structure. Homologous proteins in sequence databases are computationally modeled, to provide a resource of protein structure models complementing the experimentally solved protein structures.

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