• Title/Summary/Keyword: tryptone

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Elimination of R-Plasmid in Streptomyces bobili (YS-40) by Ethldium Bromide (Ethidium Bromide에 의한 Streptomyces bobili(YS-40)의 R-Plasmid 제거)

  • 김상달;도재호
    • Microbiology and Biotechnology Letters
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    • v.10 no.4
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    • pp.289-295
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    • 1982
  • Streptomyces bobili (YS-40) isolated from soil was tested that it had drug resistance against penicillin, cephalosporin series antibiotics and other antibiotics in the previous paper. The treatment of Streptomyces bobili, (YS-40) with ethidium bromide (EtBr), acriflavine and sodium dodecyl sulfate. (SDS) resulted in the elimination of R-plasmid from the host strain. Minimum growth inhibitory concentrations (MIC) of Hg, Ag, penicillin-G, ampicillin, chloramphenicol, oxytetracycline, streptomycin and kanamycin were found to be 15, 10, > 3, 000, > 100, > 1, 000, > 100, < 5 and < 5$\mu\textrm{g}$/$m\ell$ respectively. Among the curing agents, EtBr was proved to be the most powerful compound for the elimination of R-plasmid in the strain and the elimination rate with EtBr(10$\mu\textrm{g}$/$m\ell$) was about 98%. Optimal pH to. the elimination of R-plasmid was pH 7.0 and the R-plasmid in the cells incubated for 24 hrs was proved to be eliminated most effectively. Aerial mass color, soluble pigment formation and reverse side color were reported to be often the plasmid associated characteristics of the R-plasmid bearing bacteria. But these characteristics of the uncured and cured Streptomyces bobili, (YS-40) showed no changes in the most of the pigment formation media tested in this work.

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Effects of Growth Regulators and Organic Nitrogen Sources on the Production of Heavy Chain Immunoglobulin G in Suspension Cultures of Transgenic Tobacco Cells

  • Shin, Joong-Han;Kim, Kyoung-Heon;Kim, Tae-Hwan;Lee, James M.;Lee, Hyong-Joo
    • Journal of Microbiology and Biotechnology
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    • v.13 no.2
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    • pp.256-262
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    • 2003
  • To enhance the production of heavy chain immunoglobulin G (HC IgG) in the suspension cultures of transgenic tobacco cells (Nicotiana tabacum), the effects of adding various cytokinins (i.e., growth regulators) and organic nitrogen sources to culture media were investigated. Four different cytokinins including kinetin, isopentenyladenine (IPA), 6-benzylaminepurine (BA), and zeatin were tested with or without dichlorophenoxyacetic acid (2,4-D), which is a typical growth regulator supplemented in the standard Murashige and Skoog (MS) medium. The productivity of intracellular HC IgG was increased by 36 and $42\%$, compared to the control, especially when IPA (2 mg/l) or BA (0.2 mg/l) was added to the media in the presence of 2,4-D, respectively. In the study of organic nitrogen sources, addition of each casein hydrolysate and tryptone to the culture media at a final concentration of 0.01 and 1 g/l, respectively. increased the productivity or he IgG as much as 68 and $67\%$, respectively, in comparison with the control, which was is MS medium without supplementation of any organic nitrogen sources. This study shows that the optimization of media composition could offer significant improvements in the production of foreign proteins in the suspension cultures of transgenic plants.

Effect of culture Media on Asymbiotic Seed Germination and Those Seedling Growth of Calanthe discolor and Habenaria radiata (새우난초와 해오라비난초 종자의 기내발아와 유묘생장에 미치는 배지의 영향)

  • 정미영;정재동;지선옥
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.189-194
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    • 1998
  • The experiment was tried to identify culture media which was suitable for seed germination and seedling growth of wild orchids, Calanthe discolor and Habenaria radiata. When seeds of Calanthe discolor, which was treated with ultrasonics for 30 minutes, were sowed in Murashige and SKoog(MS) medium, germination was much more promoted than other treatments. Seedling of C. discolor grew more rapidly in 3g/L Hyponex and 2g/L tryptone(H$_3$T$_2$) medium and 3g/L Hyponex and 2g/L peptone(H$_3$P$_2$) medium, especially in H$_3$P$_4$ medium among those media, pseudobulb became more corpulent. Habenaria radiata, whose tubers were obtained from seedlings, were sprouted more vigorously in 3g/L Hyponex and 1g/L peptone(H$_3$P$_1$) medium and 1g/L Hyponex and 2g/L peptone(H$_1$P$_2$) medium; but multiplication of tubers, growth of daughter tubers and its corpulence were well established in 1g/L Hyponex and 2g/L peptone(H$_1$P$_2$) medium.

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Production of Tween 80-inducing Esterase by Acinetobacter sp. B1 Using Response Surface Methodology

  • Ma, Peiyu;Li, Yuqi;Miao, Chensi;Sun, Yunpeng;Liu, Chunhui;Li, Huijuan
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.87-95
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    • 2019
  • Esterase produced by Acinetobacter sp. B1 (strain B1) was optimized by means of one-variable-at-a-time and response surface methodologies. Results of the one-variable-at-a-time experiment showed that Tween 80 significantly increased esterase production of strain B1. The addition of Tween 80 to the culture medium increased the biomass and esterase activity of strain B1, stimulated content of total extracellular protein, and enhanced the oleic acid (C18:1) composition in the cell membrane of strain B1. The influence of eight culture variables on esterase production was evaluated by Plackett-Burman design. Results showed that Tween 80, pH, and $K_2HPO_4$ significantly affected the esterase production of strain B1. Tween 80, pH, and $K_2HPO_4$ were further optimized by central composite design. Under the optimized conditions (w/v, soluble starch 2.5%, tryptone 1.5%, Tween 80 0.8%, $K_2HPO_4$ 0.5%, NaCl 0.5%, pH 8.0, inoculum size 1%, and inoculum age 24 h), the maximum esterase activity of strain B1 was 152.13 U/ml, which was 10-fold higher than that of non-optimization after 36 h cultivation.

Optimization of Medium Composition for Biomass Production of Lactobacillus plantarum 200655 Using Response Surface Methodology

  • Choi, Ga-Hyun;Lee, Na-Kyoung;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • v.31 no.5
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    • pp.717-725
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    • 2021
  • This study aimed to optimize medium composition and culture conditions for enhancing the biomass of Lactobacillus plantarum 200655 using statistical methods. The one-factor-at-a-time (OFAT) method was used to screen the six carbon sources (glucose, sucrose, maltose, fructose, lactose, and galactose) and six nitrogen sources (peptone, tryptone, soytone, yeast extract, beef extract, and malt extract). Based on the OFAT results, six factors were selected for the Plackett-Burman design (PBD) to evaluate whether the variables had significant effects on the biomass. Maltose, yeast extract, and soytone were assessed as critical factors and therefore applied to response surface methodology (RSM). The optimal medium composition by RSM was composed of 31.29 g/l maltose, 30.27 g/l yeast extract, 39.43 g/l soytone, 5 g/l sodium acetate, 2 g/l K2HPO4, 1 g/l Tween 80, 0.1 g/l MgSO4·7H2O, and 0.05 g/l MnSO4·H2O, and the maximum biomass was predicted to be 3.951 g/l. Under the optimized medium, the biomass of L. plantarum 200655 was 3.845 g/l, which was similar to the predicted value and 1.58-fold higher than that of the unoptimized medium (2.429 g/l). Furthermore, the biomass increased to 4.505 g/l under optimized cultivation conditions. For lab-scale bioreactor validation, batch fermentation was conducted with a 5-L bioreactor containing 3.5 L of optimized medium. As a result, the highest yield of biomass (5.866 g/l) was obtained after 18 h of incubation at 30℃, pH 6.5, and 200 rpm. In conclusion, mass production by L. plantarum 200655 could be enhanced to obtain higher yields than that in MRS medium

Incubation Conditions Affecting Bacteriocin Production of Lactobacillus plantarum K11 Isolated from Dongchimi (동치미에서 분리된 Lactobacillus plantarum K11의 박테리오신 생산에 영향을 미치는 배양 조건)

  • Lim, Sung-Mee;Lee, Gun-Ja;Park, Sun-Mee;Im, Dong-Soon
    • Journal of Food Hygiene and Safety
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    • v.23 no.2
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    • pp.113-120
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    • 2008
  • The influence of incubation temperature, pH and media components on bacteriocin production by Lactobacillus plantarum K11 were investigated. The highest activity was observed in MRS broth, but no bacteriocin activity was obtained in TSB. The bacteriocin was produced from the exponential growth phase and its activity also reached a maximum in MRS broth, but then dropped after 16 hr because of degradation by extracellular proteolytic enzymes or exhaustion of medium nutrients. The optimal temperature and pH for production of bacteriocin were $37^{\circ}C$ and pH 7.0 in MRS broth, respectively. The addition of 0.5 or 1.0% glucose and $0.5{\sim}1.5%$ lactose to MRS resulted in the increase of the bacteriocin production. With 0.5% NaCl and $K_2HPO_4$, the activities were significantly higher than that of control, respectively. However, increasing nitrogen sources such as beef extract, casein, and tryptone and salts such as $NH_4PO_4$, $MgSO_47H_2O$, and $MnSO_4H_2O$, had detected a negative influence upon the bacteriocin production. Consequently, because the bacteriocin produced by L. plantarum K11 was affected by various incubation conditions, the bacteriocin activity of L. plantarum K11 applied in food as a novel starter will be dependent on environmental factors such as fermentation conditions and food ingredients.

Plant Growth Promoting Effect and Antifungal Activity of Bacillus subtilis S37-2 (Bacillus subtilis S37-2 균주의 항진균활성 및 식물생육촉진 효과)

  • Kwon, Jang-Sik;Weon, Hang-Yeon;Suh, Jang-Sun;Kim, Wan-Gyu;Jang, Kab-Yeul;Noh, Hyung-Jun
    • Korean Journal of Soil Science and Fertilizer
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    • v.40 no.6
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    • pp.447-453
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    • 2007
  • With a broad objective for the development of microbial based fertilizers, a total of 373 strains were isolated from rhizoplane and rhizosphere of pepper, tomato, lettuce, pasture, and grass. The efficacy of the isolates to augument overall plant growth was evaluated. After screening for their plant growth promotion and antagonistic properties in vitro efficient strains were further selected. The most efficient strains was characterized by 16S rRNA gene sequences and biochemical techniques and was designated as Bacillus subtilis S37-2. The strains facilitated plant growth and inhibited the plant phathogenic fungi such as Fusarium oxysporum (KACC 40037, Rhizoctonia solani (KACC 40140), and Sclerotinia sclerotiorum (KACC 40457). Pot based bioassay using lettuce as test plant was conducted by inoculating suspension ($10^5$ to $10^8cells\;mL^{-1}$) of B. subtilis S37-2 to the rhizosphere of lettuce cultivated in soil pots. Compared with non-inoculated pots, marked increase in leaf (42.3%) and root mass (48.7%) was observed in the inoculation group where the 50ml of cell mixture ($8.7{\times}10^8cells\;ml^{-1}$) was applied to the rhizosphere of letuce either once or twice. Antagonistic effects of B. subtilis S37-2 strain on S. sclerotiorum (KACC 40457) were tested. All the tested lettuce plants perished after 9 days in treatment containing only S. sclerotiorum, but only 17% of lettuce was perished in the inoculation plot. B. subtilis grew well in the TSB culture medium. The isolates grew better in yeast extracts than peptone and tryptone as nitrogen source. The growth rate was 2~4 times greater at $37^{\circ}C$ as compared with $30^{\circ}C$ incubation temperature. B. subitlis S37-2 produced $0.1{\mu}g\;ml^{-1}$ of IAA (indole 3-acetic acid) in the TSB medium containing L-tryptophan($20mg\;L^{-1}$) in 24 hours.

Effect of Amino Acids and Organic Nitrogen Sources on Cyclosporin A Fermentation by Tolypocladium inflatum (Tolypocladium inflatum을 이용한 Cyclosporin A 발효에서 아미노산과 유기질소원의 영향)

  • Kim, Jeong-Keun;Lee, Byung-Kyu;Chang, Seog-Won;Park, Yong-Deok;Rho, Yong-Taek
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.140-146
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    • 2009
  • Cyclosporin, an immunosuppressant, is a representative group of biologically active secondary metabolites produced by the fungus Tolypocladium inflatum. The amount and ratio of cyclosporin derivatives in the culture broth are an important factors for the production of cyclosporin A and the purification in the industrial process. Therefore, we studied the effect of amino acids and complex organic nitrogen sources using Tolypocladium inflatum mutants on the productivity of cyclosporin A and the ratio of cyclosporin derivatives. Overproducing mutant YHC-004 having seven times higher productivity than mother strain's could be obtained through the artificial mutation by UV irradiation. The concentration and kind of organic nitrogens and amino acids shows the profound effect on the productivity of cyclosporin A and ratio of cyclosporin derivatives. As a result, it was possible to raise the productivity and the ratio of cyclosporin A up to 3,430 mg/L and 93% respectively, but on the other hand the other cyclosporin derivatives decreased less than 2% in the culture broth.

Chitinase of Multifunctional Antagonistic Bacterium Bacillus amyloliquefaciens 7079 against Phy-tophathogenic fungi (식물병원진균을 길항하는 chitinase 생산성 생물방제균 Bacillus amyloliquefaciens 7079의 선발과 chitinase 생산조건)

  • 한옥경
    • Microbiology and Biotechnology Letters
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    • v.29 no.3
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    • pp.142-148
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    • 2001
  • An indigenous antagonistic bacterium Bacillus sp. 7079 was isolated from a local soil sampled at Kyongju area in Korea . The strain has strong antagonistic ability which was originated from multifunctional mechanisms of chitinase and antibiotic and is a powerful antagonistic biocontrol agent against red-pepper rotting fungus Phytophthora capsici and Wilt fungus Fusarium oxysporum. The chitinase might degrade the cell wasll for Fusarium species. The selected Bacilus sp. 7079 was identified as a Bacillus amyloliquefaciens 7079. The maximal production of the chitinase from B, amyloliquefaciens 7079 were obtained in chitin-yeast extract medium containing 0.7%, $K_2$$HPO_4$, $0.2KH_2$$PO_4$, 0.1% ($NH_4$)$_2$$SO_4$, 0.05% sodium cirate, 0.01% $MgSO_4$$7H_2$O, 0.1% yeast extract and 0.1% colloidal chitin after cultivation of 3 days at pH 7.0 and $30^{\circ}C$. The best carbon and nitrogen sources for the production of the chitinase from B amyloliquefaciens 7079 were determined to be 0.1% colloi- dal chitin and 0.15% proteose peptone NO 3 respectively, The antagonistic activity of B amyloliquefaciens 7079 was confirmed using P. capsici by in vivo pot test with red-pepper plant.

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Cultural Characteristics of a Biosurfactant-Producing Microorganism Pseudomonas aeruginosa F722 (Biosurfactant 생산균주 Pseudomonas aeruginosa F722의 배양특성)

  • ;;;Motoki Kubo
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.171-176
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    • 2003
  • Productivity of biosurfactant (rhamnolipid) by Pseudomonas aeuginosa F722 was investigated in the several culture conditions and culture composition. Biosurfactant production by P. aeuginosa F722 was amounted to 0.78 g/l as the result of the nitrogen sources and carbon sources without investing of optimum conditions. As for that one was investigated, biosurfactant production by P. aeruginosa F722 was amounted to 1.66 g/l. Biosurfactant production increased twofold because the composition of a modified C-medium was investigated efficiently. $NE_4$Cl or $NaNO_2$ inorganic nitrogens and yeast extract or trypton organic nitrogens were effective, but others inorganic nitrogens and organic nitrogens tested were not efficient far biosurfactant production by P. aeruginosa F722. The optimum concentration of $NH_4$Cl; inorganic nitrogen and yeast extract; organic nitrogen were 0.05% and 0.1%, respectively. In various carbon sources, others with the exception of hydrophobic property substrate (n-alkane) and hydrophilic property substrate (glucose, glycol) were not found to be effective fur biosurfactant production, and 3.0% was better in yield than other concentration of glucose. This yielded C-to-N ratios between 17 and 20. In our experiment, the highest biosurfactant production by P. aeruginosa F722 were observed in 5 days cultivation, containing glucose 3.0%, $NH_4$Cl 0.05%, and yeast extract 0.1% and C-to-N ratio was 20. Optimal pH and temperature for biosurfactant production were 7.0 and $35^{\circ}C$, respectively. Under the optimal culture conditions with glucose, biosurfactant production was amounted to 1.66 g/l. Velocity of biosurfactant production and strain growth increased after nitrogen depletion. The average surface tension of 30 mN/m after the 3 days of incubation under optimal culture condition was measured by ring tensionmeter.