• 제목/요약/키워드: M9 minimal media

검색결과 15건 처리시간 0.02초

Optimized M9 Minimal Salts Medium for Enhanced Growth Rate and Glycogen Accumulation of Escherichia coli DH5α

  • Wang, Liang;Liu, Qinghua;Du, Yangguang;Tang, Daoquan;Wise, Michael J.
    • 한국미생물·생명공학회지
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    • 제46권3호
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    • pp.194-200
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    • 2018
  • Glycogen plays important roles in bacteria. Its structure and storage capability have received more attention recently because of the potential correlations with environmental durability and pathogenicity. However, the low level of intracellular glycogen makes extraction and structure characterization difficult, inhibiting functional studies. Bacteria grown in regular media such as lysogeny broth and tryptic soy broth do no accumulate large amounts of glycogen. Comparative analyses of bacterial media reported in literature for glycogen-related studies revealed that there was no consistency in the recipes reported. Escherichia coli $DH5{\alpha}$ is a convenient model organism for gene manipulation studies with respect to glycogen. Additionally, M9 minimal salts medium is widely used to improve glycogen accumulation, although its composition varies. In this study, we optimized the M9 medium by adjusting the concentrations of itrogen source, tryptone, carbon source, and glucose, in order to achieve a balance between the growth rate and glycogen accumulation. Our result showed that $1{\times}M9$ minimal salts medium containing 0.4% tryptone and 0.8% glucose was a well-balanced nutrient source for enhancing the growth and glycogen storage in bacteria. This result will help future investigations related to bacterial physiology in terms of glycogen function.

세균 배양액과 반응한 Lactobacillus plantarum의 생장속도 및 항균 활성에 대한 연구 (Growth Rate and Antimicrobial Activity of Lactobacillus plantarum Reacting with Bacterial Culture Supernatant)

  • 하은미
    • 한국미생물·생명공학회지
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    • 제43권3호
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    • pp.187-194
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    • 2015
  • 인간의 위장관은 매우 복잡한 장내균총으로 이루어져 있으며, 장내균총은 인간의 생리 기능에 있어 매우 중요한 역할을 담당하고 있다. 특히 유아기의 장내균총의 불균형은 면역연관 질병을 일으키는 요인이며 치료 목적으로 probiotics가 유용하게 활용되고 있다. Probiotics의 대표 균종은 L. plantarum으로서 영유아기에 병원균보다 가장 먼저 정착해야 면역 질환을 예방할 수 있다. 그러나 아직 Lactobacillus가 장의 우점종으로 정착하기 위한 환경적 요인과 항생물질분비 조절에 대한 이해는 부족한 실정이다.

도로면 강우유출수의 입도분포를 고려한 여재특성 비교분석 (A Comparison of Filtering Characteristics of Various Media considering Particle Size Distribution of Road Runoff)

  • 구본진;최계운;최원석;송창수
    • 상하수도학회지
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    • 제27권3호
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    • pp.299-312
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    • 2013
  • This study investigated the contaminant loading and characteristics of particle size distributions(PSDs) in the rainfall runoff from two different sources, the pavement road and the ancillary parking lot, and then evaluated four different types of filter media(i.e., EPP, EPS, Zeolite, and Perlite) to treat runoff water. The results showed that runoff from the pavement road contains 5.6 and 20 times higher SS and Pb concentrations, respectively, than that from the parking lot. The particles smaller than $100{\mu}m$ occupied 89.8 % of runoff from the pavement road and 81.4 % of that from the parking lot by volume. The effect of the hydraulic loading, at 950 m/day filtering linar velocity and 40 cm head loss, was largest for Zeolite, followed by Perlite, EPS, and EPP. The return period of tested media calculated by the regression equation for head loss indicated that EPP has the longest life time. The average SS removal rate was similar for all media at between 84.9 % and 89 %, while the effect of various filter column heights was different, showing minimal for EPP and maximum for EPS. All filter media tested demonstrated over 95 % of SS treatment efficiency for the particles bigger than $100{\mu}m$, while for the ones smaller than $100{\mu}m$ the efficiency was in order of EPP(82.4%) > Perlite(76.1 %) > EPS(66.2 %) > Zeolite(65.2 %). The results in conclusion implies that EPP is most effective filter media for the highly contaminated fine particles from road runoff.

Structural Effects of the GXXXG Motif on the Oligomer Formation of Transmembrane Domain of Syndecan-4

  • Song, Jooyoung;Kim, Ji-Sun;Choi, Sung-Sub;Kim, Yongae
    • Bulletin of the Korean Chemical Society
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    • 제34권12호
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    • pp.3577-3585
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    • 2013
  • Syndecan-4 (heparan sulfate proteoglycan), biologically important in cell-to-cell interactions and tumor suppression, was studied through mutation of the GXXXG motif of its transmembrane domain (Syd4-TM), a motif which governs dimerization. The expression and purification of the mutant (mSyd4-TM) were optimized here to assess the function of the GXXXG motif in the dimerization of Syd4-TM. mSyd4-TM was obtained in M9 minimal media and its oligomerization was identified by SDS PAGE, Circular Dichroism (CD) spectroscopy, mass spectrometry and NMR spectroscopy. The mutant, unlike Syd4-TM, did not form dimers and was observed as monomers. The GXXXG motif of Syd-4TM was shown to be an important structural determinant of its dimerization.

Potential for the Uptake and Removal of Arsenic [As (V) and As (III)] and the Reduction of As (V) to As (III) by Bacillus licheniformis (DAS1) under Different Stresses

  • Tripti, Kumari;Sayantan, D.;Shardendu, Shardendu;Singh, Durgesh Narain;Tripathi, Anil K.
    • 한국미생물·생명공학회지
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    • 제42권3호
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    • pp.238-248
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    • 2014
  • The metalloid arsenic (Z = 33) is considered to be a significant potential threat to human health due to its ubiquity and toxicity, even in rural regions. In this study a rural region contaminated with arsenic, located at longitude $85^{\circ}$ 32'E and latitude $25^{\circ}$ 11'N, was initially examined. Arsenic tolerant bacteria from the rhizosphere of Amaranthas viridis were found and identified as Bacillus licheniformis through 16S rRNA gene sequencing. The potential for the uptake and removal of arsenic at 3, 6 and 9 mM [As(V)], and 2, 4 and 6 mM [As(III)], and for the reduction of the above concentrations of As(V) to As(III) by the Bacillus licheniformis were then assessed. The minimal inhibitory concentrations (MIC) for As(V) and As(III) was determined to be 10 and 7 mM, respectively. At 3 mM 100% As(V) was uptaken by the bacteria with the liberation of 42% As(III) into the medium, whereas at 6 mM As(V), 76% AS(V) was removed from the media and 56% was reduced to As(III). At 2 mM As(III), the bacteria consumed 100%, whereas at 6 mM, the As(III) consumption was only 40%. The role of pH was significant for the speciation, availability and toxicity of the arsenic, which was measured as the variation in growth, uptake and content of cell protein. Both As(V) and As(III) were most toxic at around a neutral pH, whereas both acidic and basic pH favored growth, but at variable levels. Contrary to many reports, the total cell protein content in the bacteria was enhanced by both As(V) and As(III) stress.

Recombinant Expression, Isotope Labeling, and Purification of Cold shock Protein from Colwellia psychrerythraea for NMR Study

  • Moon, Chang-Hun;Jeong, Ki-Woong;Kim, Hak-Jun;Heo, Yong-Seok;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • 제30권11호
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    • pp.2647-2650
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    • 2009
  • Cold shock proteins (Csps) are a subgroup of the cold-induced proteins on reduction of the growth temperature below the physiological temperature. They preferentially bind to single-stranded nucleic acids to translational regulation via RNA chaperoning. Csp plays important role in cold adaptations for the psychrophilic microorganism. Recently, Cold shock protein from psychrophilic bacteria, Colwellia psychrerythraea (CpCsp) has been identified. Three dimensional structures of a number of Csps from various microorganisms have been solved by NMR spectroscopy or X-ray crystallography, but structures of psychrophilic Csps were not studied yet. Therefore, cloning and purification protocols for further structural study of psychrophilic Csp have been optimized in this study. CpCsp was expressed in E. coli with pET-11a vector system and purified by ion exchange, size exclusion, and reverse phase chromatography. Expression and purification of CpCsp in M9 minimal media was carried out and $^{15}N$-labeled proteins with high purity over 90% was obtained. Further study will be carried out to investigate the tertiary structure and dynamics of CpCsp.

Production and Characterization of Nitrate Reductase Deficient Mutants in Petunia parviflora

  • Lee, Cheol-Hee
    • 한국자원식물학회지
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    • 제19권6호
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    • pp.706-715
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    • 2006
  • Nitrate reductase deficient (NR) mutant lines were selected indirectly by their resistance to 100mM chlorate in cell cultures of P. parviflora. A total of 585 chlorate resistant lines were confirmed by a second passage on a high concentration of chlorate. Frequency of spontaneous mutation was $9.7{\times}10^{-7}$ in 3 month old suspension-cultured cells, and in non-selective media containing amino acids as sole nitrogen source. The frequency of mutation could be increased up to 11-fold by culture for 12 months. Out of 40 randomly selected calli, 22 were fully deficient in NR. The rest of the clones contained a decreased level of NR activity. Further characterization was carried out in 13 mutant lines which were fully deficient in NR and in 5 mutant lines containing residual (0-7.0%) NR activity, as compared to wild-type cells cultured on the same medium. The $NR^-$ mutants were tentatively classified as defective in the NR apoenzyme (nia-type; 11 mutant lines including the 5 with residual NR activity) or in the molybdenum cofactor (cnx-type; 7 mutant lines) by the XDH activity. The cnx-type could be further classified into two groups. In one group (5 mutant lines) of these, the NR activity could be partially restored by nonphysiologically high (1.0mM) molybdate in the culture medium. Both types of $NR^-$ mutants were unable to grow on minimal medium containing nitrate as sole nitrogen source, but grew well on amino acids. They also proved to be extremely sensitive to the standard medium ($MSP_1$) containing nitrate and ammonium. Shoot regeneration was obtained only in the $NR^-$ mutants, which contained residual NR activity, but they so far have failed to grow into plants.

Anti-Oomycete Activity and Pepper Root Colonization of Pseudomonas plecoglossicida YJR13 and Pseudomonas putida YJR92 against Phytophthora capsici

  • Elena, Volynchikova;Ki Deok, Kim
    • The Plant Pathology Journal
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    • 제39권1호
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    • pp.123-135
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    • 2023
  • Previously, Pseudomonas plecoglossicida YJR13 and Pseudomonas putida YJR92 from a sequential screening procedure were proven to effectively control Phytophthora blight caused by Phytophthora capsici. In this study, we further investigated the anti-oomycete activities of these strains against mycelial growth, zoospore germination, and germ tube elongation of P. capsici. We also investigated root colonization ability of the bacterial strains in square dishes, including cell motility (swimming and swarming motilities) and biofilm formation. Both strains significantly inhibited mycelial growth in liquid and solid V8 juice media and M9 minimal media, zoospore germination, and germ tube elongation compared with Bacillus vallismortis EXTN-1 (positive biocontrol strain), Sphingomonas aquatilis KU408 (negative biocontrol strain), and MgSO4 solution (untreated control). In diluted (nutrient-deficient) V8 juice broth, the tested strain populations were maintained at >108 cells/ml, simultaneously providing mycelial inhibitory activity. Additionally, these strains colonized pepper roots at a 106 cells/ml concentration for 7 days. The root colonization of the strains was supported by strong swimming and swarming activities, biofilm formation, and chemotactic activity towards exudate components (amino acids, organic acids, and sugars) of pepper roots. Collectively, these results suggest that strains YJR13 and YJR92 can effectively suppress Phytophthora blight of pepper through direct anti-oomycete activities against mycelial growth, zoospore germination and germ tube elongation. Bacterial colonization of pepper roots may be mediated by cell motility and biofilm formation together with chemotaxis to root exudates.

석영 및 철피복 모래에서 박테리아 부착.탈착: 이온강도의 영향 (Adhesion and Release of Bacteria in Quartz and Iron-coated Sands: Effect of Ionic Strength)

  • 이창구;박성직;김현정;한용운;박정안;김성배
    • 대한환경공학회지
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    • 제31권4호
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    • pp.287-293
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    • 2009
  • 본 연구에서는 칼럼실험을 이용하여 석영모래와 철피복 모래에서 박테리아(Escherichia coli, Bacillus subtilis, 그리고 Staphylococcus aureus)의 부착 및 탈착에 미치는 이온강도의 영향을 조사하였다. 실험결과, 철피복 모래에서는 이온강도가 1 mM에서 100 mM로 증가함에 따라 질량회수율이 일정(E. coli = 13.7${\pm}$0.5%, B. subtilis = 9.8${\pm}$1.3%, S. aureus = 13.0${\pm}$2.1%)한 반면, 석영모래에서는 80.7%에서 45.3%로 감소하였다(S. aureus). 용출용액의 이온강도가 100 mM에서 0.1 mM로 감소함에 따라, 석영모래에서는 평균 39.1%의 박테리아 탈착이 일어났지만, 철피복 모래에서는 탈착이 관찰되지 않았다. 철피복 모래에서 관찰된 이러한 현상은 박테리아와 철피복 모래사이의 inner-sphere complexes(이온강도의 영향을 받지 않는)에 의한 결합 때문이다. 본 연구는 박테리아와 표면변형 다공성 여재의 상호작용에 대한 지식을 증진시킨다.

단일 박테리오파지를 이용한 볏짚 유래 Bacillus cereus free 스타터 컬쳐의 개발 (The Use of the Pathogen-specific Bacteriophage BCP8-2 to Develop a Rice Straw-derived Bacillus cereus-free Starter Culture)

  • 나디카 반다라;정서진;정도연;김광표
    • 한국식품과학회지
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    • 제46권1호
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    • pp.115-120
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    • 2014
  • 본 연구는 목적은 단일 박테리오파지를 이용하여 볏짚으로부터 B. cereus가 제거된 스타터 컬쳐를 개발하는 것이다. 다양한 배지 가운데 5% 콩 갈은 물(GB)이 볏짚 미생물의 증식과 박테리오파지 BCP8-2의 활성에 적합한 배지로 선정되었다. GB를 이용하여 다양한 볏짚 샘플과 BCP8-2를 함께 처리한 결과 모든 샘플들에서 총균수는 $10^9$ CFU/mL 수준으로 동일하였고, B. cereus의 경우 BCP8-2를 처리하지 않은 샘플에서 $10^7$ CFU/mL까지 증식하였고, 박테리오파지 처리군에서는 검출되지 않음을 확인하였다. 샘플에 존재하는 미생물 군집을 16S rDNA 염기서열을 바탕으로 한 DGGE와 pyrosequencing을 통해 분석한 결과 BCP8-2 처리군과 미처리군 사이에 매우 유사한 미생물 군집을 확인하였고, 더 나아가 계대 배양에 따른 미생물 군집이 안정적으로 유지됨을 확인했다. 이러한 결과들은 볏짚 등에서 특정 박테리오파지를 이용하여 특정 유해균을 제거한 컬쳐의 개발이 가능함을 보여주는 것이고, 안전한 전통장류의 생산에 기여할 것이다.