• 제목/요약/키워드: strain SK1

검색결과 57건 처리시간 0.03초

Molecular Cloning of the DNA Gyrase Genes from Methylovorus Sp. Strain SS1 and the Mechanism of Intrinsic Quinolone Resistance in Methylotrophic Bacteria

  • Kim, Kwang-Seo;Kim, Jeong Hoon;Kim, Do Yeob;Kim, Hyun Jong;Park, Sang Tae;Kim, Young Min
    • Molecules and Cells
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    • 제20권3호
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    • pp.392-400
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    • 2005
  • The genes encoding the DNA gyrase A (GyrA) and B subunits (GyrB) of Methylovorus sp. strain SS1 were cloned and sequenced. gyrA and gyrB coded for proteins of 846 and 799 amino acids with calculated molecular weights of 94,328 and 88,714, respectively, and complemented Escherichia coli gyrA and gyrB temperature sensitive (ts) mutants. To analyze the role of type II topoisomerases in the intrinsic quinolone resistance of methylotrophic bacteria, the sequences of the quinolone resistance-determining regions (QRDRs) in the A subunit of DNA gyrase and the C subunit (ParC) of topoisomerase IV (Topo IV) of Methylovorus sp. strain SS1, Methylobacterium extorquens AM1 NCIB 9133, Methylobacillus sp, strain SK1 DSM 8269, and Methylophilus methylotrophus NCIB 10515 were determined. The deduced amino acid sequences of the QRDRs of the ParCs in the four methylotrophic bacteria were identical to that of E. coli ParC. The sequences of the QRDR in GyrA were also identical to those in E. coli GyrA except for the amino acids at positions 83, 87, or 95. The $Ser^{83}$ to Thr substitution in Methylovorus sp. strain SS1, and the $Ser^{83}$ to Leu and $Asp^{87}$ to Asn substitutions in the three other methylotrophs, agreed well with the minimal inhibitory concentrations of quinolones in the four bacteria, suggesting that these residues play a role in the intrinsic susceptibility of methylotrophic bacteria to quinolones.

Kinetic and Spectral Investigations on $Ca^{2+}$ - and Sr$^{2+}$ -containing Methanol Dehydrogenases

  • Kim, Si-Wouk;Kim, Chun-Sung;Lee, Jung-Sup;Koh, Moon-Joo;Yang, Song-Suk;Duine, Johannis-A.;Kim, Young-Min
    • Journal of Microbiology
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    • 제35권3호
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    • pp.200-205
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    • 1997
  • Bothl $Ca^{2+}$ and Sr$^{2+}$-containing methanol dehydrogenases (MDH) were purified to homogeneity with yields of 48% and 42%, respectively, from Methylabacillus methanolovorus sp. strain SK5. Most of the biochemical and structural properties were similar to each other. However, some differences were found: (1) although the overall shape of the absorption spectrum of Sr$^{2+}$-MDH was very similar to that of $Ca^{2+}$-MDH, the absorption intensity originating from the cofactor in Sr$^{2+}$. MDH was higher than that in $Ca^{2+}$-MDH. Small blue shift of the maximum was also observed. These are probably due to a difference in redox state of the cofactors in $Ca^{2+}$ and Sr$^{2+}$-MDH; (2) Sr$^{2+}$-MDH was more heat-stable than $Ca^{2+}$-MDH above 56$^{\circ}C$; (3) the V$_{max}$ values for the methanol-dependent activities of Sr$^{2+}$- and $Ca^{2+}$-MDH in the presence of 3 mM KCN were 2.038 and 808 nmol/mg protein/min, respectively. In addition, the $K_{m}$ values of Sr$^{2+}$ and $Ca^{2+}$ MDH for methanol were 12 and 21 $\mu$M, respectively; (4) the endogenous activity of $Ca^{2+}$-MDH was more sensitive than that of Sr$^{2+}$-MDH in the presence of cyanide; (5) Diethyl pyrocarbonate treatment increased the enzyme activities of $Ca^{2+}$- and Sr$^{2+}$-MDH 4.2- and 1.4-folds, respectively. These results indicate that Sr$^{2+}$ stabilizes the structural conformation and enhances the activity of MDH more than $Ca^{2+}$.

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Polyethylene flow prediction with a differential multi-mode Pom-Pom model

  • Rutgers, R.P.G.;Clemeur, N.;Debbaut, B.
    • Korea-Australia Rheology Journal
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    • 제14권1호
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    • pp.25-32
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    • 2002
  • We report the first steps of a collaborative project between the University of Queensland, Polyflow, Michelin, SK Chemicals, and RMIT University, on simulation, validation and application of a recently introduced constitutive model designed to describe branched polymers. Whereas much progress has been made on predicting the complex flow behaviour of many - in particular linear - polymers, it sometimes appears difficult to predict simultaneously shear thinning and extensional strain hardening behaviour using traditional constitutive models. Recently a new viscoelastic model based on molecular topology, was proposed by McLeish and carson (1998). We explore the predictive power of a differential multi-mode version of the porn-pom model for the flow behaviour of two commercial polymer melts: a (long-chain branched) low-density polyethylene (LDPE) and a (linear) high-density polyethylene (HDPE). The model responses are compared to elongational recovery experiments published by Langouche and Debbaut (19c99), and start-up of simple shear flow, stress relaxation after simple and reverse step strain experiments carried out in our laboratory.

Methylophaga aminosulfidovorans SKI bfmo 오페론의 클로닝 및 구조 분석 (Cloning and Structural Analysis of bfmo Operon in Methylophaga aminosulfidovorans SK1)

  • 임현숙;구재환;김리현;김시욱;조은희
    • 미생물학회지
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    • 제41권1호
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    • pp.1-7
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    • 2005
  • Methylophaga aminosulfidovorans SK1 (KCTC 10323 BP)은 단일 탄소원, 질소원 그리고 에너지원으로 난분해성 화합물인 트리메틸아민을 이용할 수 있다. M. aminosulfidovorans SK1는 진핵세포의 flavin-containing monooxygenase와 유사한 유전자(bFMO)를 지니고 있으며 대장균에서 발현된 재조합 단백질은 강력한 트리메틸아민 산화활성을 보인다. 본 연구에서는 bEMO의 기능과 조절 메커니즘을 연구하기 위하여 bfmo의 상단부 및 하단부 유전자의 염기서열을 결정하였다. bfmo 상단부의 세 개의 열린해독틀은 잘 보존된 nitrate/nitrite response regulators와 methyl accepting protein 유사단백질을 암호화하였다. 하단부의 두 개의 작은 열린해독틀은 기능은 알려져 있지 않지만 진정세균계에서 잘 보존된 단백질의 일종으로 나타났다. 역전사효소 중합효소증폭반응을 통하여 여섯 개의 유전자는 세 개의 독립된 오페론으로 구성되어 있음을 확인하였다. bfmo의 상단부에 위치하는 세 개의 조절유전자는 두 개의 프로모터에서 전사되었다. 그리고 이와 독립적으로 bfmo와 두 개의 하단부 유전자가 하나의 전사단위를 이루고 있다.

실트질 모래지반의 비선형 강도 및 강성도 추정법 (Estimation of Non-linear Strength and Stiffness for Silty Sands)

  • 이경숙;김현주;이준환
    • 한국지반공학회논문집
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    • 제22권1호
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    • pp.35-44
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    • 2006
  • 일반적으로 현장지반은 대부분 일정량의 세립분을 포함하고 있으므로 세립분 함유량을 고려한 지반 거동에 관한 연구가 요구된다. 이와 더불어, 기초 지반은 하중초기단계부터 상당한 비선형 거동특성을 나타내고 있어, 응력단계에 따른 지반의 비선형성 또한 매우 중요한 고려사항이다. 본 연구에서는 일련의 실내시험을 수행하여 지반의 전단강도 및 비선형 감쇠 특성을 도출하고자 하였으며, 수정 Hyerbolic 모델을 적용하여 분석된 실트질 모래 지반의 비선형 거동특성은 실트함유량과 상대밀도에 의해 정량화되었다. 실트질 모래 시료의 응력-변형률 곡선을 도출하기 위해 세립분 함유량을 변화시켜가며 일련의 삼축압축시험이 수행되었다. 또한 비선형 특성의 정규화에 요구되는 미소변형률 구간의 초기전단탄성계수의 도출을 위해서 삼축압축시험의 시료조건과 유사한 시료에 대해 공진주시험이 수행되었다. 또한 실트함유량별로 도출된 비선형 특성치는 상대밀도의 영향이 가장 큰 것으로 나타났으며, 상대밀도 증가에 따라 파괴시의 탄성계수의 비인 f값은 감소하고, 지반강성도 감쇠율을 나타내는 9의 경우 증가하는 경향을 나타내었다. 이와 더불어 비선형 특성치를 상대밀도 $D_R$을 간극비로 환산한 절대적 간극비 $e_{sk}$에 따른 정량화 결과의 경우, $e_{sk}$의 증가에 따라 f값은 증가하고 g값은 감소하는 경향을 나타내었다.

Inhibitory Effect of Aged Petroleum Hydrocarbons on the Survival of Inoculated Microorganism in a Crude-Oil-Contaminated Site

  • Kang, Yoon-Suk;Park, Youn-Jong;Jung, Jae-Joon;Park, Woo-Jun
    • Journal of Microbiology and Biotechnology
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    • 제19권12호
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    • pp.1672-1678
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    • 2009
  • We studied the effects of aged total petroleum hydrocarbons (aged TPH) on the survival of allochthonous diesel-degrading Rhodococcus sp. strain YS-7 in both laboratory and field investigations. The aged TPH extracted from a crude-oil-contaminated site were fractionized by thin-layer chromatography/flame ionization detection (TLC/FID). The three fractions identified were saturated aliphatic (SA), aromatic hydrocarbon (AH), and asphaltene-resin (AR). The ratio and composition of the separated fractions in the aged TPH were quite different from the crude-oil fractions. In the aged TPH, the SA and AH fractions were reduced and the AR fraction was dramatically increased compared with crude oil. The SA and AH fractions (2 mg/l each) of the aged TPH inhibited the growth of strain YS-7. Unexpectedly, the AR fraction had no effect on the survival of strain YS-7. However, crude oil (1,000 mg/l) did not inhibit the growth of strain YS-7. When strain YS-7 was inoculated into an aged crude-oil-contaminated field and its presence was monitored by fluorescent in situ hybridization (FISH), we discovered that it had disappeared on 36 days after the inoculation. For the first time, this study has demonstrated that the SA and AH fractions in aged TPH are more toxic to an allochthonous diesel-degrading strain than the AR fraction.

repABC- Type Replicator Region of Megaplasmid pAtC58 in Agrobacterium tumefaciens C58

  • LEE KO-EUN;PARK DAE-KYUN;BAEK CHANG-HO;HWANG WON;KIM KUN-SOO
    • Journal of Microbiology and Biotechnology
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    • 제16권1호
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    • pp.118-125
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    • 2006
  • The region responsible for replication of the megaplasmid pAtC58 in the nopaline-type Agrobacterium tumefaciens strain C58 was determined. A derivative ofa Co1E1 vector, pBluscript SK-, incapable of autonomous replication in Agrobacterium spp, was cloned with a 7.6-kb Bg1II-HindIII fragment from a cosmid clone of pAtC58, which contains a region adjacent to the operon for the utilization of deoxyfructosyl glutamine (DFG). The resulting plasmid conferred resistance to carbenicillin on the A. tumefaciens strain UIA5 that is a plasmidfree derivative of C58. The plasmid was stably maintained in the strain even after consecutive cultures for generations. Analysis of nested deletions of the 7.6-kb fragment showed that a 4.3-kb BglII-XhoI region sufficiently confers replication of the derivative of the ColE1 vector on UIA5. The region comprises three ORFs, which have high homologies with repA, repB, and repC of plasm ids in virulent Agrobacterium spp. including pTiC58, pTiB6S3, pTi-SAKURA, and pRiA4b as well as those of symbiotic plasmids from Rhizobium spp. Phylogenie analysis showed that rep genes in pAtC58 are more closely related to those in pRiA4 than to pTi plasmids including pTiC58, suggesting that the two inborn plasmids, pTiC58 and pAtC58, harbored in C58 evolved from distinct origins.

양어 사료첨가제로서의 유산균 발효 개똥쑥의 항산화 및 항균활성 (Antioxidant and Antibacterial Activities of Lactobacillus-fermented Artemisia annua L. as a Potential Fish Feed Additive)

  • 이아란;우개민;강수경;한성구;이봉주;김수기
    • 생명과학회지
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    • 제27권6호
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    • pp.652-660
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    • 2017
  • 최근 양어에서는 전염병, 환경오염, 어분 가격 상승 등으로 어려움을 가지고 있다. 본 연구는 쑥에서 분리한 L. plantarum SK3494를 이용하여 전통 약초인 개똥쑥을 발효시켜 in vitro 생리활성을 측정하고 어류용 사료첨가제로서의 가능성을 연구하였다. 개똥쑥의 발효시 유산균수는 9.38 log10 CFU/ml이며, pH는 4.1로 나타났다. 발효개똥쑥의 항산화활성은 DPPH 측정과 MAC-T 세포 내 과산화물 감소 실험에서 뛰어난 효과를 보였다. 발효개똥쑥은 어류 병원균인Photobacterium damselae subsp. damselae와 Vibrio ichthyoenteri에 대하여 강한 항균활성을 나타내었다. 따라서 유산균을 이용한 개똥쑥 발효물은 양식용 사료첨가제로써 가능성을 가지고 있는 것으로 판단되었다.

Fatigue Crack Growth Behavior of NR and HNBR Based Vulcanizates with Potential Application to Track Pad for Heavy Weight Vehicles

  • Kim, Wonho;Kim, Minyoung;Chang, Young-Wook;Shin, Jung-Eun;Bae, Jong-Woo
    • Macromolecular Research
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    • 제11권2호
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    • pp.73-79
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    • 2003
  • Generally, field performance of elastomeric track pad components has been poor, especially for the medium to heavy tonnage tracked vehicles, which are operated on the hilly cross-country course. The service life of these track pad, is affected not only by the terrain and environmental conditions but also by the speed, cornering, braking, weight of the vehicle, and the track tread design. In this research, modulus, tearing energy, and the rate of crack propagation of vulcanizates are evaluated by changing base materials to improve the service time of track pad. By increasing the contents of carbon black, modulus, tearing energy, and fatigue crack growth resistance of vulcanizates improved. Compared with the NR vulcanizate, the HNBR vulcanizate had a higher value of tearing energy. The rate of crack propagation of vulcanizates using smaller size carbon black was slower than that using larger size carbon black. When the HNBR was blended with the ZSC, the tearing energy of the vulcanizates was a little reduced because of the high modulus but the crack propagation rate was reduced significantly. In the relation between the crack propagation rate and the strain energy release rate, though up to 100% strain were applied to specimens, the slope on the log scale ($\beta$) varied between 1.72 and 2.3 with the kind of elastomer.

Overexpression of Escherichia coli Thiol Peroxidase in the Periplasmic Space

  • Kim, Sung-Jin;Cha, Mee-Kyung;Kim, Il-Han;Kim, Ha-Kun
    • Journal of Microbiology and Biotechnology
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    • 제8권1호
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    • pp.92-95
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    • 1998
  • Overproduction of Escherichia coli thiol peroxidase in the periplasmic space was achieved by locating the appropriate gene on a downstream region of the strong T7 promoter. E. coli strain BL21 carrying the recombinant plasmid pSK-TPX was induced by IPTG, lysed, and analyzed by SDS-polyacrylamide gel electrophoresis. A large amount of the overexpressed thiol peroxidase was located in the periplasmic space. A homogeneous thiol peroxidase was obtained from E. coli osmotic shock fluid by simple one-step gel permeation chromatography.

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