• 제목/요약/키워드: strain characterization and identification

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세포 외 중성 단백질분해효소를 생산하는 Pichia anomala CO-1의 분리 동정 및 효소 특성 (Isolation of the Protease-producing Yeast Pichia anomala CO-1 and Characterization of Its Extracellular Neutral Protease)

  • 김지연
    • 생명과학회지
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    • 제29권10호
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    • pp.1126-1135
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    • 2019
  • 세포 외로 단백질분해효소를 생산하는 효모 균주 CO-1을 대나무 부산물에서 분리하였다. CO-1은 원형 또는 타원형($3.1-4.0{\times}3.8-4.4{\mu}m$)으로, 생장을 위한 최적 온도는 $30^{\circ}C$, 초기 pH는 4.0이었다. 그리고 최대 15.0% (w/v)의 NaCl과 9.0%(v/v)의 ethanol 농도에서 생장하였다. 형태적, 생리 생화학적 특성 및 18S rRNA 유전자 염기서열을 통한 계통분석을 이용하여 동정을 실시한 결과 Pichia anomala로 판명되었다. P. anomala CO-1 단백질분해효소를 부분 정제한 결과 수율은 7.2%였으며, 정제 전에 비해 약 14.6배 정제되었다. Zymogram으로 측정한 효소의 분자량은 약 30 kDa으로 확인되었다. 본 균주는 배지 중에 탄소원과 질소원, 무기염으로 1.0%(w/v) CMC와 1.0%(w/v) yeast extract, 0.3%(w/v) $MnSO_4$를 사용하였을 경우 가장 높은 단백질분해효소 활성을 나타내었다. P. anomala CO-1이 생산하는 단백질분해효소의 최적 활성 pH와 온도는 각각 7.0과 $30^{\circ}C$였다. 또한 본 효소는 pH 4.0-10.0에서 75%의 안정성을 나타내었으며, $65^{\circ}C$에서 1시간 가열하여도 60% 전후의 활성을 유지하였다. 균주의 효소 생산은 생육과 비례하였으며 대수증식기 후반에 최대의 효소 생산을 나타내었다.

Identification and Characterization of Ligninolytic Enzyme by Serratia marcescens HY-5 isolated from the Gut of Insect

  • 김기덕;신동하;손광희;박호용
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2002년도 생물공학의 동향 (X)
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    • pp.473-476
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    • 2002
  • A lignin degradation bacteria, symbiotic bacteria was isolated from the gut of Sympetrum depressiusculum and tested for its lignin degrading activity using lignin model compounds and related aromatic compounds. The strain was identified as Serratia marcescens HY-5 based on the 165 rDNA, cellular fatty acid composition, biochemical and physiological characteristics. S. marcescens showed 40-50% lignin degrading activity in the media that contained vaillin, guaiacol and dealkaline lignin. S. marcescens showed three ligninase activities [Jaccase, lignin peroxidase(LiP) and Manganase peroxidase(MnP)]. Addition of dealkaline lignin to the basal media increased about 6fold of laccase activity. Vanillic acid or vanillin increase 1.3fold of MnP activity and p-coumaric acid increased 12fold of LiP activity which added to the basal medium.

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Purification and Characterization of a Protease Produced by a Planomicrobium sp. L-2 from Gut of Octopus vulgaris

  • Liu, Qing;Sun, Shujing;Piao, Meizi;Yang, Ji Young
    • Preventive Nutrition and Food Science
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    • 제18권4호
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    • pp.273-279
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    • 2013
  • Protease widely exists in the digestive tract of animals and humans, playing a very important role in protein digestion and absorption. In this study, a high protease-producing strain Planomicrobium sp. L-2 was isolated and identified from the digestive tract of Octopus variabilis. The strain was identified by physiological and biochemical experiments and 16S rDNA sequences analysis. A protease was obtained from the strain Planomicrobium sp. L-2 through ammonium sulfate precipitation, dialysis and enrichment, DEAE-Sephadex A50 anion-exchange chromatography, and Sephadex G-100 gel chromatography. The molecular weight and properties of the protease were characterized, including optimum temperature and pH, thermal stability, protease inhibitions and metal ions. According to our results, the protease from Planomicrobium sp. L-2 strain designated as F1-1 was obtained by three-step separation and purification from crude enzyme. The molecular weight of the protease was 61.4 kDa and its optimum temperature was $40^{\circ}C$. The protease F1-1 showed a broad pH profile for casein hydrolysis between 5.0~11.0. No residual activity was observed after incubation for 40 min at $60^{\circ}C$ and 60 min at $50^{\circ}C$. F1-1 protease was inhibited by $Mn^{2+}$, $Hg^{2+}$, $Pb^{2+}$, $Zn^{2+}$, and $Cu^{2+}$ ions, as well as PMSF, indicating that the protease F1-1 was a serine protease. Additionally, research basis provided by this study could be considered for industrial application of octopus intestinal proteases.

Isolation and characterization of a new Methanoculleus bourgensis strain KOR-2 from the rumen of Holstein steers

  • Battumur, Urantulkhuur;Lee, Manhee;Bae, Gui Sek;Kim, Chang-Hyun
    • Asian-Australasian Journal of Animal Sciences
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    • 제32권2호
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    • pp.241-248
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    • 2019
  • Objective: To isolate and identify new methanogens from the rumen of Holstein steers in Korea. Methods: Representative rumen contents were obtained from three ruminally cannulated Holstein steers ($793{\pm}8kg$). Pre-reduced media were used for the growth and isolation of methanogens. Optimum growth temperature, pH, and sodium chloride (NaCl) concentration as well as substrate utilization and antibiotic tolerance were investigated to determine the physiological characteristics of the isolated strain. Furthermore, the isolate was microscopically studied for its morphology. Polymerase chain reaction of 16S rRNA and mcrA gene-based amplicons was used for identification. Results: One strain designated as KOR-2 was isolated and found to be a non-motile irregular coccus with a diameter of 0.2 to $0.5{\mu}m$. KOR-2 utilized $H_2+CO_2$ and formate but was unable to metabolize acetate, methanol, trimethylamine, 2-propanol, and isobutanol for growth and methane production. The optimum temperature and pH for the growth of KOR-2 were $38^{\circ}C$ and 6.8 to 7.0, respectively, while the optimum NaCl concentration essential for KOR-2 growth was 1.0% (w/v). KOR-2 tolerated ampicillin, penicillin G, kanamycin, spectromycin, and tetracycline. In contrast, the cell growth was inhibited by chloramphenicol. Phylogenetic analysis of 16S rRNA and mcrA genes revealed the relatedness between KOR-2 and Methanoculleus bourgensis. Conclusion: Based on the physiological and phylogenetic characteristics, KOR-2 was thought to be a new strain within the genus Methanoculleus and named Methanoculleus bourgensis KOR-2.

Parasporin-4, A Novel Cancer Cell-killing Protein Produced by Bacillus thuringiensis

  • Inouye, Kuniyo;Okumura, Shiro;Mizuki, Eiichi
    • Food Science and Biotechnology
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    • 제17권2호
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    • pp.219-227
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    • 2008
  • Bacillus thuringiensis was isolated as a pathogen of the sotto disease of silkmoth larvae about a hundred years ago. Since then, this bacterium has attracted attentions of not only insect pathologists but also many other scientists who are interested in its strong and specific insecticidal activity. This has led to the recent worldwide development of B. thuringiensis-based microbial insecticides and insect-resistant transgenic plants, as well as a landmark discovery of par asp orin, a cancer cell-specific cytotoxin produced by B. thuringiensis. In this review, we describe examination of interaction between inclusion proteins of B. thuringiensis and brush border membrane of insects using a surface plasmon resonance-based biosensor, identification and characterization of parasporin-4, the latest parasporin produced by the B. thuringiensis A1470 strain, and an effective method for preparing the parasporin-4 from inclusion bodies expressed in the recombinant Escherichia coli cells.

In vitro Characterization of Bacteriocin Produced by Lactic Acid Bacteria Isolated from Nem Chua, a Traditional Vietnamese Fermented Pork

  • Pilasombut, Komkhae;Rumjuankiat, Kittaporn;Ngamyeesoon, Nualphan;Duy, Le Nguyen Doan
    • 한국축산식품학회지
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    • 제35권4호
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    • pp.473-478
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    • 2015
  • The aim of this study was to screen and In vitro characterize the properties of bacteriocin produced by lactic acid bacteria isolated from Vietnamese fermented pork (Nem chua). One hundred and fifty LAB were isolated from ten samples of Nem chua and screened for bacteriocin-producing lactic acid bacteria. Antimicrobial activity of bacteriocin was carried out by spot on lawn method against both gram positive and gram negative bacteria. One isolate, assigned as KL-1, produced bacteriocin and showed inhibitory activity against Lactobacillus sakei, Leuconostoc mesenteroides and Enterococcus faecalis. To characterize the bacteriocin-producing strain, optimum temperature, incubation period for maximum bacteriocin production and identification of bacteriocin-producing strain were determined. It was found that the optimum cultivation temperature of the strain to produce the maximum bacteriocin activity (12,800 AU/mL) was obtained at 30℃. Meanwhile, bacteriocin production at 6,400 AU/mL was found when culturing the strain at 37℃ and 42℃. The isolate KL-1 was identified as L. plantarum. Antimicrobial activity of cell-free supernatant was completely inhibited by proteolytic enzyme of trypsin, alpha-chymotrypsin and proteinase K. Bacteriocin activity was stable at high temperature up to 100℃ for 10 min and at 4℃ storage for 2 d. However, the longer heating at 100℃ and 4℃ storage, its activity was reduced.

Characterization of a New Leuconostoc Species Isolated from Fresh Garlic

  • Lee, Se-Hi;Choi, Jong-Hoon;Kim, Youn-Soon;Kyung, Kyu-Hang
    • Food Science and Biotechnology
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    • 제14권3호
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    • pp.416-419
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    • 2005
  • Unknown bacterium isolated from garlic was characterized using phenotypic methods, phylogenetic analysis, DNA-DNA hybridization, and cultural methods. The strain was identified as typical leuconostoc; Gram-positive, non-sporeforming, heterofermentative, catalase-negative and spherical. Although its 16S rRNA gene sequence showed high homology to Leuconostoc argentinum DSM $8581^T$(99.8%), DNA-DNA hybridization experiments indicated it represents novel genomic species in the genus Leuconostoc. The garlic-specific leuconostoc was more resistant to antimicrobial activity of garlic compared to other common laboratory lactic acid bacteria, and was even stimulated by low concentrations (1-2%) of garlic extract supplemented in trypticase soy broth. Growth stimulation was concentration-dependent when tested with residual aqueous layer after solvent extraction of fresh whole garlic extract.

Identification and Characterization of a New Strain of the Unicellular Green Alga Dunaliella salina (Teod.) from Korea

  • Polle, Jurgen E.W.;Struwe, Lena;Jin, Eon-Seon
    • Journal of Microbiology and Biotechnology
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    • 제18권5호
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    • pp.821-827
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    • 2008
  • The unicellular green alga Dunaliella salina is a halotolerant eukaryotic organism. Its halophytic properties provide an important advantage for open pond mass cultivation, since D. salina can be grown selectively. D. salina was originally described by E. C. Teodoresco in 1905. Since that time, numerous isolates of D. salina have been identified from hypersaline environments on different continents. The new Dunaliella strain used for this study was isolated from the salt farm area of the west coastal side of South Korea. Cells of the new strain were approximately oval- or pear-shaped (approximately $16-24\;{\mu}m$ long and $10-15\;{\mu}m$ wide), and contained one pyrenoid, cytoplasmatic granules, and no visible eyespot. Although levels of $\beta$-carotene per cell were relatively low in cells grown at salinities between 0.5 to 2.5 M NaCl, cells grown at 4.5 M NaCl contained about a ten-fold increase in cellular levels of $\beta$-carotene, which demonstrated that cells of the new Korean strain of Dunaliella can overaccumulate $\beta$-carotene in response to salt stress. Analysis of the ITS1 and ITS2 regions of the new Korean isolate showed that it is in the same clade as D. salina. Consequently, based on comparative cell morphology, biochemistry, and molecular phylogeny, the new Dunaliella isolate from South Korea was classified as D. salina KCTC10654BP.

Sine sweep effect on specimen modal parameters characterization

  • Roy, Nicolas;Violin, Maxime;Cavro, Etienne
    • Advances in aircraft and spacecraft science
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    • 제5권2호
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    • pp.187-204
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    • 2018
  • The sine sweep base excitation test campaign is a major milestone in the process of mechanical qualification of space structures. The objectives of these vibration tests are to qualify the specimen with respect to the dynamic environment induced by the launcher and to demonstrate that the spacecraft FE model is sufficiently well correlated with the test specimen. Dynamic qualification constraints lead to performing base excitation sine tests using a sine sweep over a prescribed frequency range such that at each frequency the response levels at all accelerometers, load cells and strain gages is the same as the steady state response. However, in practice steady state conditions are not always satisfied. If the sweep rate is too high the response levels will be affected by the presence of transients which in turn will have a direct effect on the estimation of modal parameters. A study funded by ESA and AIRBUS D&S was recently carried out in order to investigate the influence of sine sweep rates in actual test conditions. This paper presents the results of this study along with recommendations concerning the choice of methods.

음식물류폐수에 특이적 응집제를 생성하는 미생물의 분리, 동정 및 응집특성 (Isolation, Identification, and Characterization of Microorganisms which Possess the Flocculating Activity to Food Wastewater)

  • 정명희;정두영;김영준
    • 유기물자원화
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    • 제15권4호
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    • pp.125-130
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    • 2007
  • 다양한 환경으로부터 음식물류폐수에 응집특성을 보이는 미생물 6종을 분리하였다. 동정한 결과, 이들은 각각 Bacillus pumilus, Enterobacter sp., Pantotea agglomerans, Bacillus licheniformis, 그리고 두 종류의 Bacillus sp. 균주들로 밝혀졌다. 분리된 미생물중 Enterobacter sp.(YK102), Bacillus sp.(YK103), Pantotea agglomerans (YK104) 등, 3균주의 경우, 대조군으로 사용된 분양균주 Zoogloea ramigera와 비교하여 카올린에 대한 응집율이 모두 8배 이상 높은 것으로 나타났다. 음식물류폐수에 대한 응집실험에서는 Enterobacter sp.(YK102) 균주가 가장 높은 응집능을 보였으며, 대조군으로 사용된 Pseudomonas fluorescens보다 약 2.5배 높은 것으로 나타났다.

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