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Sphingopyxis granuli sp. nov., a $\beta$-Glucosidase-Producing Bacterium in the Family Sphingomonadaceae in $\alpha$-4 Subclass of the Proteobacteria

  • Kim Myung Kyum;Im Wan Taek;Ohta Hiroyuki;Lee Myung Jin;Lee Sung Taik
    • Journal of Microbiology
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    • 제43권2호
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    • pp.152-157
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    • 2005
  • Strain Kw07$^T$, a Gram-negative, non-spore-forming, rod-shaped bacterium, was isolated from granules in an Up-flow Anaerobic Sludge Blanket (UASB) bioreactor used in the treatment of brewery waste­water. 16S rRNA gene sequence analysis revealed that strain Kw07T belongs to the a-4 subclass of the Proteobacteria, and the highest degree of sequence similarity was determined to be to Sphingopyxis macrogoltabida IFO 15033T (97.8%). Chemotaxonomic data revealed that strain Kw07T possesses a quinone system with the predominant compound Q-I0, the predominant fatty acid C,s:, OJ7c, and sphingolipids, aU of which corroborated our assignment ofthe strain to the Sphingopyxis genus. The results of DNA-DNA hybridization and physiological and biochemical tests clearly demonstrated that strain Kw07T represents a distinct species. Based on these data, Kw07T (= KCTC 12209T = NBRC 100800T) should be classified as the type strain for a novel Sphingopyxis species, for which the name Sphingopyxis granuli sp. novo has been proposed.

홍조류로부터 신규 한천분해미생물 Alteromonas macleodii subsp. GNUM08120의 분리 및 동정 (Isolation and Characterization of a Novel Agar Degrading Bacterium, Alteromonas macleodii subsp. GNUM08120, from Red Macroalgae)

  • 지원재;임주현;박다연;김무찬;김창준;장용근;홍순광
    • 한국미생물·생명공학회지
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    • 제41권1호
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    • pp.8-16
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    • 2013
  • An agar-hydrolyzing marine bacterium, strain GNUM08120, was isolated from Sargassum fulvellum collected from Yeongil bay of East Sea of Korea. The isolate was Gram-negative, aerobic, motile with single polar flagellum, and grew at 1-10% NaCl, pH 5.0-8.0, and $15-37^{\circ}C$. G+C content and the predominant respiratory quinone were 46.13 mol% and Q-8, respectively. The major cellular fatty acids were Summed feature 3 (24.5%), $C_{16:0}$ (21.7%), and $C_{18:1}{\omega}7c$ (12.5%). Based on 16S rRNA gene sequence similarity and DNA-DNA hybridization analyses, strain GNUM08120 was identified as a novel subspecies of Alteromonas macleodii, designated Alteromonas macleodii subsp. GNUM08120. Production of agarase by strain GNUM08120 was likely repressed by the effect of carbon catabolite repression caused by glucose. The crude agarase prepared from 12-h culture broth of strain GNUM08120 exhibited an optimum pH and temperature for agarase activity at 7.0 and $40^{\circ}C$, respectively. The crude enzyme produced (neo)agarobiose, (neo)agarotetraose, and (neo)agarohexaose as the hydrolyzed product of agarose.

Staphylococcus epidermidis urease의 정제 및 생화학적 특성에 관한 연구 (Purification and Characterization of the Staphylococcus epidermidis Urease)

  • 민선희;이만형
    • 생명과학회지
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    • 제17권4호
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    • pp.581-586
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    • 2007
  • 본 연구에서는 피부상재균이며 기회병원균이기도 한 Staphylococcus epidermidis ATCC12228로부터 urease효소를 4단계 크로마토그라피 방법을 사용하여 1,127배 정제하고 그 생화학적인 특성을 규명하였다. 정제된 urease 효소는 SDS-PACE 전기영동분석 및 gel-filtration 크로마토그라피를 이용한 천연분자량 분석결과, 67, 16.1 및 12.7 kDa의 3개 subunit가 3량체로 회합되어 존재하는 것으로 나타났으며 catalytic unit 당 2.2개의 니켈 원소를 함유하는 것으로 측정되었다. 정제된 효소의 비활성은 993.8 U/mg, $K_m$값은 8.5mM로 각각 산출되었다.

Ramlibacter ginsenosidimutans sp. nov., with Ginsenoside-Converting Activity

  • Wang, Liang;An, Dong-Shan;Kim, Song-Gun;Jin, Feng-Xie;Kim, Sun-Chang;Lee, Sung-Taik;Im, Wan-Taek
    • Journal of Microbiology and Biotechnology
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    • 제22권3호
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    • pp.311-315
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    • 2012
  • A novel ${\beta}$-proteobacterium, designated BXN5-$27^T$, was isolated from soil of a ginseng field of Baekdu Mountain in China, and was characterized using a polyphasic approach. The strain was Gram-staining-negative, aerobic, motile, non-spore-forming, and rod shaped. Strain BXN5-$27^T$ exhibited ${\beta}$-glucosidase activity that was responsible for its ability to transform ginsenoside $Rb_1$ (one of the dominant active components of ginseng) to compound Rd. Phylogenetic analysis based on 16S rRNA gene sequences showed that this strain belonged to the family Comamonadaceae; it was most closely related to Ramlibacter henchirensis $TMB834^T$ and Ramlibacter tataouinensis$TTB310^T$ (96.4% and 96.3% similarity, respectively). The G+C content of the genomic DNA was 68.1%. The major menaquinone was Q-8. The major fatty acids were $C_{16:0}$, summed feature 4 (comprising $C_{16:1}$ ${\omega}7c$ and/or iso-$C_{15:0}$ 2OH), and $C_{17:0}$ cyclo. Genomic and chemotaxonomic data supported the affiliation of strain BXN5-$27^T$ to the genus Ramlibacter. However, physiological and biochemical tests differentiated it phenotypically from the other established species of Ramlibacter. Therefore, the isolate represents a novel species, for which the name Ramlibacter ginsenosidimutans sp. nov. is proposed, with the type strain being BXN5-$27^T$ (=DSM $23480^T$ = LMG $24525^T$ = KCTC $22276^T$).

Rheinheimera aquatica sp. nov., Antimicrobial Activity-Producing Bacterium Isolated from Freshwater Culture Pond

  • Chen, Wen-Ming;Lin, Chang-Yi;Young, Chiu-Chung;Sheu, Shih-Yi
    • Journal of Microbiology and Biotechnology
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    • 제20권10호
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    • pp.1386-1392
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    • 2010
  • A bacterial strain designated GR5$^T$, previously isolated from a freshwater culture pond in Taiwan while screening for bacteria for antimicrobial compounds, was characterized using a polyphasic taxonomic approach. Strain GR5$^T$ was found to be Gram-negative, aerobic, greenish-yellow colored, rod-shaped, and motile by means of a single polar flagellum. Growth occurred at $10-40^{\circ}C$ (optimum, $35^{\circ}C$), pH 7.0-8.0 (optimum pH 8.0), and with 0-2.0% NaCl (optimum, 0.5-1.0%). The major fatty acids were $C_{16:1}{\omega}7c$(36.3%), $C_{16:0}$(16.6%), $C_{12:0}$ 3-OH (12.5%), and $C_{18:1}{\omega}7c$(9.1%). The major respiratory quinone was Q-8, and the DNA G+C content of the genomic DNA was 51.9 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain GR5$^T$ belongs to the genus Rheinheimera, where its most closely related neighbors are Rheinheimera texasensis A62-14B$^T$ and Rheinheimera tangshanensis JA3-B52$^T$ with sequence similarities of 98.1% and 97.5%, respectively, and the sequence similarities to any other recognized species within Gammaproteobacteria are less than 96.5%. The mean level of DNA-DNA relatedness between strain GR5$^T$ and R. texasensis A62-14B$^T$, the strain most closely related to the isolate, was $26.5{\pm}7.6%$. Therefore, based on the phylogenetic and phenotypic data, strain GR5$^T$ should be classified as a novel species, for which the name Rheinheimera aquatica sp. nov. is proposed. The type strain is GR5$^T$ (=BCRC 80081$^T$=LMG 25379$^T$).

Isolation of a Novel Freshwater Agarolytic Cellvibrio sp. KY-YJ-3 and Characterization of Its Extracellular ${\beta}$-Agarase

  • Rhee, Young-Joon;Han, Cho-Rong;Kim, Won-Chan;Jun, Do-Youn;Rhee, In-Ku;Kim, Young-Ho
    • Journal of Microbiology and Biotechnology
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    • 제20권10호
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    • pp.1378-1385
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    • 2010
  • A novel agarolytic bacterium, KY-YJ-3, producing extracellular agarase, was isolated from the freshwater sediment of the Sincheon River in Daegu, Korea. On the basis of Gram-staining data, morphology, and phylogenetic analysis of the 16S rDNA sequence, the isolate was identified as Cellvibrio sp. By ammonium sulfate precipitation followed by Toyopearl QAE-550C, Toyopearl HW-55F, and MonoQ column chromatographies, the extracellular agarase in the culture fluid could be purified 120.2-fold with a yield of 8.1%. The specific activity of the purified agarase was 84.2 U/mg. The molecular mass of the purified agarase was 70 kDa as determined by dodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimal temperature and pH of the purified agarase were $35^{\circ}C$ and pH 7.0, respectively. The purified agarase failed to hydrolyze the other polysaccharide substrates, including carboxymethyl-cellulose, dextran, soluble starch, pectin, and polygalacturonic acid. Kinetic analysis of the agarose hydrolysis catalyzed by the purified agarase using thin-layer chromatography showed that the main products were neoagarobiose, neoagarotetraose, and neoagarohexaose. These results demonstrated that the newly isolated freshwater agarolytic bacterium KY-YJ-3 was a Cellvibrio sp., and could produce an extracellular ${\beta}$-agarase, which hydrolyzed agarose to yield neoagarobiose, neoagarotetraose, and neoagarohexaose as the main products.

Polyphasic Assignment of a Highly Proteolytic Bacterium Isolated from a Spider to Serratia proteamaculans

  • Kwak, Jang-Yul;Lee, Dong-Hun;Park, Youn-Dong;Kim, Seung-Bum;Maeng, Jin-Soo;Oh, Hyun-Woo;Park, Ho-Yong;Bae, Kyung-Sook
    • Journal of Microbiology and Biotechnology
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    • 제16권10호
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    • pp.1537-1543
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    • 2006
  • A bacterial strain named HY-3 that produces a highly active extracellular protease was isolated from the digestive tract of a spider, Nephila clavata. The bacterium was a Gram-negative, oxidase-negative, catalase-positive, nonhalophilic, nitrate-reducing, facultative anaerobe. Transmission and scanning electron microscopies demonstrated that the isolate was non-spare-forming, straight, rod-shaped, and motile by peritrichous flagella. The G+C content of the DNA was 57.0 mol%. The isoprenoid quinone type was ubiquinone with 8 isoprene units (Q-8). The morphological and biochemical characteristics including the predominant fatty acid and phospholipids profiles placed the isolate HY-3 in the family Enterobacteriaceae. Further biochemical characterization and phylogenetic studies including determination of an almost complete 16S ribosomal DNA sequence suggested that the bacterium was closely related to the genus Serratia. DNA-DNA hybridization analysis revealed that this extracellular protease-producing strain belongs to Serratia proteamaculans, which is also known far its association with insects.

곤충세포주에서 누에신 단백질의 발현 및 성상구명 (Characterization and Expression of Antibacterial Protein Gene, Nuecin)

  • 윤은영;구태원;황재삼;김상현;강석우;김근영;진병래
    • 한국잠사곤충학회지
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    • 제44권2호
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    • pp.64-68
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    • 2002
  • 본 연구는 곤충 유전자를 이용한 항세균성 펩타이드 생산 및 농업용 소재로서의 응용에 관한 연구로서 항세균성 단백질 누에신 유전자를 베큘로 바이러스 발현계(BEVS)를 이용하여 곤충세포주에서 발현한 후 누에신 단백질의 농업용 소재로서의 가능성을 모색하기 위해 농작물을 가해하는 감자 고추의 무름병을 일으키는 Pectobacterium carotovorum subsp. carotovorum, 가지 및 고추의 풋마름병을 일으키는 Ralstonia solanacearum, 양송이 버섯의 세균성 갈색 무늬 병을 일으키는 Pseudomonas tolaasii 및 무와 배추의 검은썩음병을 일으키는 Xanthomonas campestris pv. campestris 에 대해서 항세균 활성을 관찰하였다. 그 결과 Pectobacterium carotovorum subsp. carotovorum, Ralstonia solanacearum 및 Pseudomonas tolaasii에 대해 높은 활성을 나타내었으며 Xanthomonas campestris pv. campestris에는 활성을 나타내지 않았다. Ion exchange 및 gel filtration chromatography 를 수행하여 약 20 kDa의 성숙 누에신 단백질을 순수 분리하여 pH 및 온도에 대한 안정성을 조사한 결과, pH 2~12 완충액에서 30분간 처리하였을 때에도 항세균 활성이 그대로 유지되었고 10$0^{\circ}C$에서 2시간 처리시에는 활성이 안정되었으며 4시간 처리시에도 80% 정도로 유지됨을 확인함으로써 누에신 단백질은 pH 및 온도에 대한 안정성이 있음을 확인할 수 있었다.

Caenimonas aquaedulcis sp. nov., Isolated from Freshwater of Daechung Reservoir during Microcystis Bloom

  • Le, Ve Van;Ko, So-Ra;Lee, Sang-Ah;Kang, Mingyeong;Oh, Hee-Mock;Ahn, Chi-Yong
    • Journal of Microbiology and Biotechnology
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    • 제32권5호
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    • pp.575-581
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    • 2022
  • A Gram-stain-negative, white-coloured, and rod-shaped bacterium, strain DR4-4T, was isolated from Daechung Reservoir, Republic of Korea, during Microcystis bloom. Strain DR4-4T was most closely related to Caenimonas terrae SGM1-15T and Caenimonas koreensis EMB320T with 98.1% 16S rRNA gene sequence similarities. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strain DR4-4T and closely related type strains were below 79.46% and 22.30%, respectively. The genomic DNA G+C content was 67.5%. The major cellular fatty acids (≥10% of the total) were identified as C16:0, cyclo C17:0, summed feature 3 (C16:1ω7c and/or C16:1ω6c), and summed feature 8 (C18:1ω7c and/or C18:1ω6c). Strain DR4-4T possessed phosphatidylethanolamine, diphosphatidylglycerol, and phosphatidylglycerol as the main polar lipids and Q-8 as the respiratory quinone. The polyamine profile was composed of putrescine, cadaverine, and spermidine. The results of polyphasic characterization indicated that the isolated strain DR4-4T represents a novel species within the genus Caenimonas, for which the name Caenimonas aquaedulcis sp. nov. is proposed. The type strain is DR4-4T (=KCTC 82470T =JCM 34453T).

Variovorax terrae sp. nov. Isolated from Soil with Potential Antioxidant Activity

  • Woo, Chae Yung;Kim, Jaisoo
    • Journal of Microbiology and Biotechnology
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    • 제32권7호
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    • pp.855-861
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    • 2022
  • A white-pigmented, non-motile, gram-negative, and rod-shaped bacterium, designated CYS-02T, was isolated from soil sampled at Suwon, Gyeonggi-do, Republic of Korea. Cells were strictly aerobic, grew optimally at 20-28℃ and hydrolyzed Tween 40. Phylogenetic analysis based on 16S rRNA gene sequence indicated that strain CYS-02T formed a lineage within the family Comamonadaceae and clustered as members of the genus Variovorax. The closest members were Variovorax guangxiensis DSM 27352T (98.6% sequence similarity), Variovorax paradoxus NBRC 15149T (98.5%), and Variovorax gossypii JM-310T (98.3%). The principal respiratory quinone was Q-8 and the major polar lipids contain phosphatidylethanolamine (PE), phosphatidylethanolamine (PG), and diphosphatidylglycerol (DPG). The predominant cellular fatty acids were C16:0, summed feature 3 (C16:1ω7c and/or C16:1ω6c) and summed feature 8 (C18:1ω7c and/or C18:1ω6c). The DNA GC content was 67.7 mol%. The ANI and dDDH values between strain CYS-02T and the closest members in the genus Variovorax were ≤ 79.0 and 22.4%, respectively, and the AAI and POCP values between CYS-02T and the other related species in the family Comamonadaceae were > 70% and > 50%, respectively. The genome of strain CYS-02T showed a putative terpene biosynthetic cluster responsible for antioxidant activity which was supported by DPPH radical scavenging activity test. Based on genomic, phenotypic and chemotaxonomic analyses, strain CYS-02T was classified into a novel species in the genus Variovorax, for which the name Variovorax terrae sp. nov., has been proposed. The type strain is CYS-02T (= KACC 22656T = NBRC 00115645T).