• Title/Summary/Keyword: M9 minimal media

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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.
    • Microbiology and Biotechnology Letters
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    • v.46 no.3
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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.

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

  • Ha, Eun-Mi
    • Microbiology and Biotechnology Letters
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    • v.43 no.3
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    • pp.187-194
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    • 2015
  • In this study, L. plantarum, when reacting with the culture media of potential pathogenic bacteria, exhibited an increase in growth rate and antimicrobial activity. In order to examine the characteristics and the nature of the reaction with the bacteria, this study carried out experiments involving culturing the test bacteria in M9 minimal media. Subsequently, the supernatant was incrassated by the decompression-drying method. Through colony forming unit assay, it was confirmed that L. plantarum had the function of growth inhibition to various bacteria. After culturing L. plantarum with bacterial media, the growth rate of L. plantarum was measured by absorbance (OD600), the results showed that the growth rate (E. coli treatment group: OD600 = 0.848, S. typhimurium treatment group: OD600 = 0.848) increased, as compared with the non-treated control group (OD600 = 0.48). In contrast, the concentrate itself did not induce the growth of L. plantarum. These results were observed as a universal phenomenon of the Lactobacillus species. Moreover, the increase in antimicrobial activity was observed in L. plantarum, which reacted with the culture media of E. coli and S. typhimurium, through a disc diffusion assay, and the result of growth inhibition against various bacteria was induced. Finally, based on the analysis results of the characteristics of bacteria culture media, which increased the growth rate of L. plantarum and antibacterial activity, the bacterial media had a tolerance for catabolic enzymes, pH 2−8 and heat. Therefore, this substance can be said to be a small molecule which is highly stable under various conditions.

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

  • Koo, Bonjin;Choi, Gyewoon;Choi, Weonsuk;Song, Changsoo
    • Journal of Korean Society of Water and Wastewater
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    • v.27 no.3
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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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    • v.34 no.12
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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.
    • Microbiology and Biotechnology Letters
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    • v.42 no.3
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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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    • v.30 no.11
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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
    • Korean Journal of Plant Resources
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    • v.19 no.6
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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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    • v.39 no.1
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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.

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

  • Lee, Chang-Gu;Park, Seong-Jik;Kim, Hyon-Chong;Han, Yong-Un;Park, Jeong-Ann;Kim, Song-Bae
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.4
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    • pp.287-293
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    • 2009
  • This study investigated the influence of ionic strength on the adhesion and release of bacteria (Escherichia coli, Bacillus subtilis, and Staphylococcus aureus) in quartz and iron-coated sands using column experiments. Results show that the mass recovery remained constant (E. coli = 13.7${\pm}$0.5%, B. subtilis = 9.8${\pm}$1.3%, S. aureus = 13.0${\pm}$2.1%) in iron-coated sand while it decreased from 80.7 to 45.3% (S. aureus) in quartz sand with increasing ionic concentrations from 1 to 100 mM. As the ionic concentrations of leaching solution was lowered from 100 to 0.1 mM, average 39.1% of bacterial detachment was quantified from quartz sand, but no bacterial release was observed in iron-coated sand. The phenomenon observed in iron-coated sand can be attributed to the inner-sphere complexes between bacteria and coated sand, which have minimal effect from ionic strength. This study improves our knowledge regarding the bacterial interaction with surface-modified porous media.

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

  • Bandara, Nadeeka;Chung, Seo-Jin;Jeong, Do-Youn;Kim, Kwang-Pyo
    • Korean Journal of Food Science and Technology
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    • v.46 no.1
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    • pp.115-120
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    • 2014
  • The purpose of this study was to develop a rice straw-derived Bacillus cereus (B. cereus)-free starter culture for traditional soybean fermented products using a B. cereus-specific bacteriophage, BCP8-2. To determine the optimal medium that supports the growth of rice straw-derived microorganisms and BCP8-2 activity, 5 different culture media were tested. The 5% ground bean (GB) medium was selected for further study. No B. cereus was detected in the BCP8-2-treated rice straw in GB medium, whereas B. cereus at a level of $10^7$ CFU/mL was recovered in the no-phage control. The total bacterial count reached approximately $10^9$ CFU/mL regardless of phage addition. When the 16S rRNA sequence-based microbial community was monitored using denaturing gradient gel electrophoresis (DGGE) and pyrosequencing, a similar microbial community was observed in the phage-treated and control samples. In conclusion, we demonstrate that phage can be used to prepare a rice straw-derived B. cereus-free starter culture with minimal effect on natural microflora.