• 제목/요약/키워드: microbial characterization

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Microbial Community Profiling in cis- and trans-Dichloroethene Enrichment Systems Using Denaturing Gradient Gel Electrophoresis

  • Olaniran, Ademola O.;Stafford, William H.L.;Cowan, Don A.;Pillay, Dorsamy;Pillay, Balakrishna
    • Journal of Microbiology and Biotechnology
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    • 제17권4호
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    • pp.560-570
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    • 2007
  • The effective and accurate assessment of the total microbial community diversity is one of the primary challenges in modem microbial ecology, especially for the detection and characterization of unculturable populations and populations with a low abundance. Accordingly, this study was undertaken to investigate the diversity of the microbial community during the biodegradation of cis- and trans-dichloroethenes in soil and wastewater enrichment cultures. Community profiling using PCR targeting the l6S rRNA gene and denaturing gradient gel electrophoresis (PCR-DGGE) revealed an alteration in the bacterial community profiles with time. Exposure to cis- and trans-dichloroethenes led to the disappearance of certain genospecies that were initially observed in the untreated samples. A cluster analysis of the bacterial DGGE community profiles at various sampling times during the degradation process indicated that the community profile became stable after day 10 of the enrichment. DNA sequencing and phylogenetic analysis of selected DGGE bands revealed that the genera Acinetobacter, Pseudomonas, Bacillus, Comamonas, and Arthrobacter, plus several other important uncultured bacterial phylotypes, dominated the enrichment cultures. Thus, the identified dominant phylotypes may play an important role in the degradation of cis- and trans-dichloroethenes.

대두접종제(大豆接種劑) 개발(開發)을 위한 우수근류균(優秀根瘤菌)의 선발(選拔) 및 plasmid 특성(特性) (Selection of R. japonicum Strains for Developing Soybean Inoculant and Plasmid Characterization)

  • 김창진;김성훈;이윤;유익동;민태익
    • Applied Biological Chemistry
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    • 제28권3호
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    • pp.149-155
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    • 1985
  • 전국(全國) 223개(個) 지역(地域)에서 수집(蒐集)한 두과작물(豆科作物)의 근류(根瘤)에서 총(總) 590주(株)의 근류균(根瘤菌)을 분리(分離)하여 숙주특이성(宿主特異性)에 따라, R. japonicum 218종(種), R. phaseoli 101종(種), R. trifolii 97종(種), R. meliloti 4종(種), R. leguminosarum 1종(種), Rhizobium species 101종(種), 미분류(未分類)된 균주(菌株) 159종(種)으로 분류(分類)하였다. 분리(分離)된 R. japonicum 218종(種)의 근류형성능(根瘤形成能)과 질소고정능(窒素固定能)을 비교(比較)하여. R. japonicum R-138, R-168, R-214 균주(菌株)를 우수균주(優秀菌株)로 하였다. 분리(分離) 선발(選拔)된 R. japonicum 중(中) 질소고정능(窒素固定能)이 우수(優秀)한 균주(菌株)의 plasmid를 확인(確認)한 결과(結果), fast-growing group 균주(菌株)들은 $1{\sim}4$개(個)의 거대(巨大) plasmid를 함유(含有)하고 있었으며 그 분자량(分子量)은 약(約) $35{\sim}300{\;}Md$ 정도(程度)이었다. 반면 대부분(大部分)의 slow-growing group 균주(菌株)들은 plasmid가 검출(檢出)되지 않았다.

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Microbial Rhodopsins: Genome-mining, Diversity, and Structure/Function

  • Jung, Kwang-Hwan;Vishwa Trivedi;Yang, Chii-Shen;Oleg A. Sineschekov;Elena N. Spudich;John L. Spudich
    • Journal of Photoscience
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    • 제9권3호
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    • pp.45-48
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    • 2002
  • Microbial rhodopsins, photoactive 7-transmembrane helix proteins that use retinal as their chromophore, were observed initially in the Archaea and appeared to be restricted to extreme halophilic environments. Our understanding of the abundance and diversity of this family has been radically transformed by findings over the past three years. Genome sequencing of cultivated microbes as well as environmental genomics have unexpectedly revealed archaeal rhodopsin homologs in the other two domains of life as well, namely Bacteria and Eucarya. Organisms containing these homologs inhabit such diverse environments as salt flats, soil, freshwater, and surface and deep ocean waters, and they comprise a broad phylogenetic range of microbial life, including haloarchaea, proteobacteria, cyanobacteria, fungi, and algae. Analysis of the new microbial rhodopsins and their expression and structural and functional characterization reveal that they fulfill both ion transport and sensory functions in various organisms, and use a variety of signaling mechanisms. We have obtained the first crystallographic structure for a photosensory member of this family, the phototaxis receptor sensory rhodopsin II (SRII, also known as phoborhodopsin) that mediates blue-light avoidance by the haloarchaeon Natronobacterium pharaonis. The structure obtained from x-ray diffraction of 3D crystals prepared in a cubic lipid phase reveals key features responsible for its spectral tuning and its sensory function. The mechanism of SRII signaling fits a unified model for transport and signaling in this widespread family of phototransducers.

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Recent Development of Rapid and Automation Technology for Food Microbiological Examination

  • Hiroshi Kurata
    • 한국식품위생안전성학회:학술대회논문집
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    • 한국식품위생안전성학회 1996년도 제11회 학술대회 및 정기총회 - 식품의 위생 안전성에 관한 최근 연구 동향
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    • pp.33-33
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    • 1996
  • Interests in the field of rapid methods and automation in microbiology have been growing steadily on an international scale in recent years. International meetings concerned this problem have been held in elsewhere in the world countries since the past twenty years. But, unfortunately in the field of microbial examination in food hygiene, this problem have not yet been developed so much as in the field of clinical microbiology. Today, I would like to introduce you here present aspects of rapid and automation technologies, those which are manly carrying in milk and meats industries. My illustration will be given recent improved technologies using automatic apparatus and instruments along with process of microbial count procedure. Recent direct microbiological counting system (ChemeScan \ulcorner) as real time ultrasensitive analysis created by Cheminex Ltd., France is now most evolutional instrument to provide direct microbial counts, down to one cell, within 30 minutes. The results from these evaluations how a good correlation between the ChemScan system and the standard plate count method. This system will be successful application for not only in the field of pharmacology but also food microbiology. In addition, current identification of microbes by sophisticated instruments suitable for food microbiology, one of which Biology is manual system (BIOLOG\ulcorner), provides reference-level capability at a modes price. For the manual system, the color reactions in the microplate are read by eye and manually keyed into personal computer. Species identification appears on the computer screen within seconds, along with biotype patterns, a list of closely related species, and other useful statistics. In present this is useful application for microbial ecology and epidemiological survey. RiboPrinter system newly produced by DuPont is now focusing among microbiologists in the world, and is one of the biggest microbial characterization system using a DNA-based approach. The technology analyzer is bacterial culture for its genetic fingerprint or riboprint pattern. Finally Bio-cellTracer system for automatic measurement of fungal growth and Fukitori-Maseter, a Surface Hygiene Monitoring Kit by using swabe procedure in food processing environment are briefly illustrated in this presentation.

Characterization of a Thermophilic Lignocellulose-Degrading Microbial Consortium with High Extracellular Xylanase Activity

  • Zhang, Dongdong;Wang, Yi;Zhang, Chunfang;Zheng, Dan;Guo, Peng;Cui, Zongjun
    • Journal of Microbiology and Biotechnology
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    • 제28권2호
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    • pp.305-313
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    • 2018
  • A microbial consortium, TMC7, was enriched for the degradation of natural lignocellulosic materials under high temperature. TMC7 degraded 79.7% of rice straw during 15 days of incubation at $65^{\circ}C$. Extracellular xylanase was effectively secreted and hemicellulose was mainly degraded in the early stage (first 3 days), whereas primary decomposition of cellulose was observed as of day 3. The optimal temperature and initial pH for extracellular xylanase activity and lignocellulose degradation were $65^{\circ}C$ and between 7.0 and 9.0, respectively. Extracellular xylanase activity was maintained above 80% and 85% over a wide range of temperature ($50-75^{\circ}C$) and pH values (6.0-11.0), respectively. Clostridium likely had the largest contribution to lignocellulose conversion in TMC7 initially, and Geobacillus, Aeribacillus, and Thermoanaerobacterium might have also been involved in the later phase. These results demonstrate the potential practical application of TMC7 for lignocellulosic biomass utilization in the biotechnological industry under hot and alkaline conditions.

Characterization of Calcium-Activated Bifunctional Peptidase of the Psychrotrophic Bacillus cereus

  • Kim Jong-Il;Lee Sun-Min;Jung Hyun-Joo
    • Journal of Microbiology
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    • 제43권3호
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    • pp.237-243
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    • 2005
  • The protease purified from Bacillus cereus JH108 has the function of leucine specific endopeptidase. When measured by hydrolysis of synthetic substrate (N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide), the enzyme activity exhibited optimal activity at pH 9.0, $60^{\circ}C$. The endopeptidase activity was stimulated by $Ca^{++},\;Co^{++},\;Mn^{++},\;Mg^{++},\;and\;Ni^{++}$, and was inhibited by metal chelating agents such as EDTA, 1,10-phenanthroline, and EGTA. Addition of serine protease inhibitor, PMSF, resulted in the elimination of the activity. The endopeptidase activity was fully recovered from the inhibition of EDTA by the addition of 1 mM $Ca^{++}$, and was partially restored by $Co^{++}\;and\;Mn^{++}$, indicating that the enzyme was stabilized and activated by divalent cations and has a serine residue at the active site. Addition of $Ca^{++}$ increased the pH and heat stability of endopeptidase activity. These results show that endopeptidase requires calcium ions for activity and/or stability. A Lineweaver-Burk plot analysis indicated that the $K_m$ value of endopeptidase is 0.315 mM and $V_{max}$ is 0.222 ) is $0.222\;{\mu}mol$ of N-succinyl-Ala-Ala-Pro-Leu-p-nitroanilide per min. Bestatin was shown to act as a competitive inhibitor to the endopeptidase activity.

Isolation and Genomic Characterization of the T4-Like Bacteriophage PM2 Infecting Pectobacterium carotovorum subsp. carotovorum

  • Lim, Jeong-A;Lee, Dong Hwan;Heu, Sunggi
    • The Plant Pathology Journal
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    • 제31권1호
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    • pp.83-89
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    • 2015
  • In order to control Pectobacterium carotovorum subsp. carotovorum, a novel virulent bacteriophage PM2 was isolated. Bacteriophage PM2 can infect 48% of P. carotovorum subsp. carotovorum and 78% of P. carotovorum subsp. brasilliensis but none of atrosepticum, betavasculorum, odoriferum and wasabiae isolates had been infected with PM2. PM2 phage belongs to the family Myoviridae, and contains a large head and contractile tail. It has a 170,286 base pair genome that encodes 291 open reading frames (ORFs) and 12 tRNAs. Most ORFs in bacteriophage PM2 share a high level of homology with T4-like phages including IME08, RB69, and JS98. Phylogenetic analysis based on the amino acid sequence of terminase large subunits confirmed that PM2 is classified as a T4-like phage. It contains no integrase- or no repressor-coding genes related to the lysogenic cycle, and lifestyle prediction using PHACT software suggested that PM2 is a virulent bacteriophage.

미생물학적 위해성 평가 (Microbial Risk Assessment)

  • 이건형
    • 미생물학회지
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    • 제37권2호
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    • pp.101-108
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    • 2001
  • 환경에서 병원성 미생물의 발생에 관련된 자료를 분석하는 일은 잠재적 보건 위해성을 판단하고 그에 따른 적절한 조치를 취하는데 종종 결정적인 역할을 한다. 이러한 일들은 과거에는 주로 정성적이고, 주관적인 방법으로 이루어 졌지만, 최근에는 정량적 위해성 평가(Quantitative Risk Assessment: QRA)로 이루어지는데, 이러한 QRA는 병원성 미생물로부터의 감염이나 발병, 또는 치사율 등의 위해성을 정량적인 방법으로 표현하는 접근법이다. 따라서, 이와 같은 정보는 정책 결정권자가 위해성의 확대와 비용부담, 그리고 적절한 조치를 취했을 때 기대할 수 있는 이점을 결정하는데 보다 더 잘 이용할 수 있다. 본 문에서는 위해성 분석의 과제에 대한 일반적인 배경을 살펴보고 문제해결 과정에서 미생물 위해성 평가가 어떻게 활용 가능한지를 살펴보았다.

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Cultivation-Dependent and -Independent Characterization of Microbial Community Producing Polyhydroxyalkanoates from Raw Glycerol

  • Ciesielski, Slawomir;Pokoj, Tomasz;Klimiuk, Ewa
    • Journal of Microbiology and Biotechnology
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    • 제20권5호
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    • pp.853-861
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    • 2010
  • High substrate costs decrease the profitability of polyhydroxyalkanoates (PHAs) production, and thus low-cost carbon substrates coming from agricultural and industrial residuals are tested for the production of these biopolymers. Among them, crude glycerol, formed as a by-product during biodiesel production, seems to be the most promising source of carbon. The object of this study was to characterize the mixed population responsible for the conversion of crude glycerol into PHAs by cultivation-dependent and -independent methods. Enrichment of the microbial community was monitored by applying the Ribosomal Intergenic Spacer Analysis (RISA), and the identification of community members was based on 16S rRNA gene sequencing of cultivable species. Molecular analysis revealed that mixed populations consisted of microorganisms affiliated with four bacterial lineages: ${\alpha}$, ${\gamma}$-Proteobacteria, Actinobacteria, and Bacteroides. Among these, three Pseudomonas strains and Rhodobacter sp. possessed genes coding for polyhydroxyalkanoates synthase. Comparative analysis revealed that most of the microorganisms detected by direct molecular analysis were obtained by the traditional culturing method.

Purification and Characterization of a Thermostable Laccase from Trametes trogii and Its Ability in Modification of Kraft Lignin

  • Ai, Ming-Qiang;Wang, Fang-Fang;Huang, Feng
    • Journal of Microbiology and Biotechnology
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    • 제25권8호
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    • pp.1361-1370
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    • 2015
  • A blue laccase was purified from a white rot fungus of Trametes trogii, which was a monomeric protein of 64 kDa as determined by SDS-PAGE. The enzyme acted optimally at a pH of 2.2 to 4.5 and a temperature of 70℃ and showed high thermal stability, with a half-life of 1.6 h at 60℃. A broad range of substrates, including the non-phenolic azo dye methyl red, was oxidized by the laccase, and the laccase exhibited high affinity towards ABTS and syringaldazine. Moreover, the laccase was fairly metal-tolerant. A high-molecular-weight kraft lignin was effectively polymerized by the laccase, with a maximum of 6.4-fold increase in weight-average molecular weight, as demonstrated by gel permeation chromatography. Notable structural changes in the polymerized lignin were detected by Fourier transform infrared spectroscopy and 1H NMR spectroscopy. This revealed an increase in condensed structures as well as carbonyl and aliphatic hydroxyl groups. Simultaneously, phenolic hydroxyl and methoxy groups decreased. These results suggested the potential use of the laccase in lignin modification.