• Title/Summary/Keyword: growth medium optimization

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Antimicrobial activities of Burkholderia sp. strains and optimization of culture conditions (Burkholderia sp. OS17의 항균활성 증진을 위한 배양최적화)

  • Nam, Young Ho;Choi, Ahyoung;Hwang, Buyng Su;Chung, Eu Jin
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.428-435
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    • 2018
  • In this study, we isolated and identified bacteria from freshwater and soil collected from Osang reservoir, to screen antimicrobial bacteria against various pathogenic bacteria. 38 strains were isolated and assigned to the class Proteobacteria (22 strains), Actinobacteria (7 strains), Bacteroidets (6 strains), and Firmicutes (3 strains) based on 16S rRNA gene sequence analysis. Among them, strain OS17 showed a good growth inhibition against 5 methicillin-resistant Staphylococcus aureus subsp. aureus strains and Bacillus cereus, Bacillus subtilis, Filobasidium neoformans. As a result of the 16S rRNA gene sequence analysis, strain OS17 show the high similarity with Burkholderia ambifaria $AMMD^T$, B. diffusa $AM747629^T$, B. tettitorii $LK023503^T$ 99.8%, 99.7%, 99.6%, respectively. We investigated cell growth and antimicrobial activity according to commercial culture medium, temperature, pH for culture optimization of strain OS17. Optimal conditions for growth and antimicrobial activity in strain OS17 were found to be: YPD medium, $35^{\circ}C$ and pH 6.5. When the strain was cultured in LB, NB, TSB, R2A media at $20^{\circ}C$ and $25^{\circ}C$, the antimicrobial activity did not show. Culture filtrate of strain OS17 showed antimicrobial activity against 5 MRSA strains, Bacillus cereus, Bacillus subtilis, and Filobasidium neoformans with inhibition zones from 2 to 8 mm. Optimal reaction time was 48 h in YPD medium, 100 rpm and 0.3 vvm in 2 L-scale fed-batch fermentation process for antimicrobial activity. Culture optimization of strain OS17 can be improved on antimicrobial activity. Therefore, the antimicrobial activity of Burkholderia sp. OS17 had potential as antibiotics for pathogens including MRSA.

High cell density cultivation of Bacillus sp.

  • Lee, Baek-Seok;Chae, Won-Bok;Jo, Jae-Hui;Choe, Gi-Hyeon;Kim, Yeong-Beom;Choe, Seong-Won;Kim, Eun-Gi
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.290-293
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    • 2001
  • In this study, media optimization by statistically designed experiments stimulated an increase in cell growth of Bacillus sp. during batch cultivation. Plackett-Surman design method selected 3 components among 7 components of production medium. Box-Behnken design method calculated the optimum concentration of selected components by Plackett-Surman design. In the optimized medium, viable cell number increased 2 times. Addition of antifoam effected the cell growth depending on the type of antifoam Vegetable oil, are a carbon source and an antifoam. increased cell growth and controlled foaming

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Optimazation of Submerged Culture Conditions for Exo-Biopolymer Production by Paecilomyces japonica

  • Bae, Jun-Tae;Sinha, Jayanta;Park, Jong-Pil;Song, Chi-Hyun;Yun, Jong-Won
    • Journal of Microbiology and Biotechnology
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    • v.10 no.4
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    • pp.482-487
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    • 2000
  • Optimization of submerged culture conditions for the production of exo-biopolymer from Paecilomyces japonica ws studied. Maltose, yeast extract, and potassium phosphate were the most suitable sources of carbon, nitrogen, and inorganic salt, respectively, for both production of the exo-biopolymer and mycelial growth. The optimal culture conditions in a flask culture were pH 5.0, $25^{\circ}C$, and 150 rpm in a medium containing (as in g/l) 30 maltose, 6 yeast extruct, 2 polypeptone, $0.5{\;}K_3HPO_4,{\;}0.2{\;}KH_2PO_4,{\;}0.2{\;}MnSO_4{\cdot}5H_2O,{\;}0.2{\;}MgSO_4{\cdot}7H_2O$. Exo-biopolymer production and mycelial growth in the above suggested medium were significantly increased in a 2.5-1 jar fermentor, where the maximum biopolymer concentration was 8 g/l. The morphological changes of the mycelium in the submerged culture were observed within pH ranges from 4.0 to 9.0; i.e., growth of the filamentous form was optimal at culture pHs of 5.0 and 6.0, whereas pellet was formed at other pHs.

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Optimization of Culture Conditions and Analysis of Plasmid Stability of a Transformant Bacillus subtilis for Cytidine Deaminase Production

  • Kim, Soo-Hyun;Song, Bang-Ho;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.1 no.2
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    • pp.116-120
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    • 1991
  • The transformant Bacillus subtilis ED213 carrying the pSO100 which cloned the cdd gene encoding cytidine deaminase (cytidine /2'-deoxycytidine aminohydrolase, EC 3.5.4.5, CDase) originated from wild type B. subtilis was cultivated in Spizizen minimal medium (SMM). To overcome poor expression of the cdd gene in SMM medium, the medium compositions and growth conditions were optimized. The optimized medium compositions and growth conditions were cytidine concentration of 80 mg/l, glycerol of 25 g/l, and $(NH_4)_2SO_4$ of 10 g/l, along with $37^{\circ}C$ and pH 7.0. The intracellular CDase production was increased 3 times from 1,000 unit/ml to 3,200 unit/ml, and extracellular CDase also increased from nearly undetectable amounts to 1,500 unit/ml. The cytidine concentration was signified as the most critical compositional factor for overproduction of CDase by increasing the cell density mainly in culture broth. The plasmids were more stable in cells that were grown in original SMM medium with stability of 90% compared to those grown in optimized SMM medium with stability of 80% after 48 hours cultivation. The most active amplification of plasmid was occurred in the logarithmic phase, which showed a value around four times higher than the initial copy number. In the exponential phase, the CDase production was closely related to the plasmid copy number along with the cell density. However, it was not accorded with cell density at the stationary phase.

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In Vitro Proliferation Model of Helicobacter pylori Required for Large-Scale Cultivation

  • Oh, Heung-Il;Lee, Heung-Shick;Kim, Kyung-Hyun;Paek, Se-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.10 no.3
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    • pp.367-374
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    • 2000
  • The composition of dissolved gases and nutrients in a liquid medium were determined for establishment of the optimum conditions for in vitro culture of Helicobacter pylori. A microaerobic condition facored by the organism was prepared by adjusting the partial pressure of the gas, agitation speed, and viscosity of the medium. The gaseous concentrations were controlled by utilizing CampyPak Plus that reduced oxygen while augmenting carbon dioxide. Agitation of the broth facilitated the oxygen transfer to the cells, yet inhibited the growth at high rates. An increase of viscosity in the medium repressed the culture although this variable was relatively insignificant. The chemical constituents of the liquid broth were examined to establish an economic model for H. pylori cultivation. The microbe required a neutral pH for optimum growth, and yet was also able to proliferate in an acidic condition, presumably by releasing the acidity-modulating enzyme, urease. Cyclodextrin and casamino acid were investigated as growth enhancers in place of serum, while yeast extract unexpectedly inhibited the cells. A low concentration of glucose, the unique carbon source for the organism, increased the cell density, yet high concentrations resulted in an adverse effect. Under optimally dissolved gas conditions, the cell concentration in brucella broth supplemented with serum substitutes and glucose reached $1.6{\times}10^8$ viable cells/ml which was approximately 50% higher than that obtained in the liquid medium added with only cyclodextrin or serum.

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Composition Optimization of Cabbage Extract Medium for Cell Growth of Lactobacillus plantarum (식물성 배지에서 Lactobacillus plantarum의 배양을 위한 배지 최적화)

  • Jeong, Eun Ji;Moon, Dae Won;Oh, Joon Suk;Moon, Jin Seok;Eom, Hyun Ju;Choi, Hye Sun;Kim, Chang Sup;Han, Nam Soo
    • KSBB Journal
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    • v.27 no.6
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    • pp.347-351
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    • 2012
  • This study was conducted to optim ize the composition of CEM (cabbage extract medium) and cryoprotectants on the growth of Lactobacillus plantarum, a probiotics growing in plant and milk. For this, we analyzed the growth characteristics of Lb. plantarum in CEM and subsequently optimized the medium composition by addition of carbon, nitrogen sources and buffering agents. Among carbon sources, glucose showed the best result to increase the cell density after dilution of CEM. When 0.5% yeast extract and 1% soy peptone were supplemented in the diluted CEM, Lb. plantarum grew up to the maximum cell density. Addition of buffering agents in CEM was not significantly effective to increase the cell density. Meanwhile, addition of 12% skim milk, 5% sucrose and 0.5% glycerol showed a cryoprotective effect against cell damage of Lb. plantarum during freeze drying process showing high survival rate after 150 days. This optimized CEM can be used for economical production of bacterial cells particularly originated from a plant-related ecosystem.

Identification of characterization and statistical optimization of medium constituent for Bacillus subtilis SCJ4 isolated from Korean traditional fermented food (전통 장류 유래 Bacillus subtilis SCJ4의 특성확인 및 통계학적 방법을 이용한 배양조건 최적화)

  • Jeong, Su-Ji;Yang, Hee-Jong;Jeong, Seong-Yeop;Jeong, Do-Youn
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.48-60
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    • 2015
  • 612 strains isolated from Korean traditional fermented food in Sunchang and their investigated biochemical characterization and ability of biogenic amines non-producing. We selected the SCJ4 having various activity by measurement of extracellular enzyme, antioxidant and antimicrobial activities. Selected strain SCJ4 by 16S rRNA sequencing and biochemical characterization was named Bacillus subtilis SCJ4. And then, we investigated cell growth of SCJ4, and optimized of culture medium constituents using response surface methodology as statistically method. Response surface methodology used Plackett-Burman experimental design for screening of medium constituent. Tryptone, peptone and $MgSO_4$ as medium constituent improving cell growth selected. In order to find out optimal concentration on each constituent, we carried out central composite design. Consequently, optimized concentrations of tryptone, peptone and $MgSO_4$ were predicted to be 15.35 g/L, 12.235 g/L, and 3.5 g/L respectively. Through the model verification, we confirmed about 1.28-fold improvement of the dried cell weight from 0.8767 g/L to 1.1222 g/L when compared to basal medium.

Optimization for Mycelial Growth and Inhibitory Effect on Nitric Oxide Production of Cordyceps nutans Pat. (노린재동충하초의 배양 최적화 및 NO 생성 저해 효과)

  • Lee, Ki-Man;Lee, Geum-Seon;Nam, Sung-Hee;Lim, Sung-Cil;Kang, Tae-Jin
    • Korean journal of applied entomology
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    • v.50 no.4
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    • pp.307-314
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    • 2011
  • Cordyceps (vegetable wasp and plant worm), an entomopathogenic fungi, has been used as a herbal medicine in Asian countries since ancient times. Cordyceps nutans is common but there is little research on this species. This study investigated the optimal culture conditions of C. nutans and the inhibitory effect on nitric oxide (NO) production in RAW 264.7 cell treated culture broth. The optimal conditions for the mycelial growth were $25^{\circ}C$ and pH 7.0-8.0. Mycelial growth was highest on mushroom complete medium (MCM), V8 juice agar (V8A), and yeast malt dextrose (YMD) medium. Mycelial growth on mushroom minimal medium (MMM) did not occur, so nutrient source was essential. Dextrose and sucrose as carbon sources, and ammonium citrate as a nitrogen source were satisfactory for mycelial growth. Cytotoxicity of C. nutans culture broth was not found in RAW 264.7 cells. C. nutans culture broth suppressed NO production of lipopolysaccharide (LPS)-stimulated RAW 264.7 cell in a dose-dependent manner. Thus, our results provided the optimal conditions for cultivation of C. nutans and showed that C. nutans may have excellent physiological activities.

Growth Optimization of Photorhabdus luminescens Isolated from Entomopathogenic Nematode Heterorhabditis bacteriophora (병원성 선충 Heterorhabditis bacteriophora에서 분리된 공생 박테리아 Photorhabdus luminescens의 생장조건)

  • Yoo, Sun Kyun;Randy Gaugler;Christopher W. Brey
    • Microbiology and Biotechnology Letters
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    • v.29 no.2
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    • pp.104-109
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    • 2001
  • The yield of infective juveniles of Heterorhabditis bacteriophora (Tf strain) in vitro monoxenic liquid culture was improved significantly as the amount of symbiont biomass, Photorhabdus sp. strain Tf, increased. To investigate the influence of abiotic factors on the growth and biomass production of Photorhabdus sp. strain Tf, triplicate flask cu1tmes were performed. The optinal temperature and medium pH for the growth of Photorhahdus sp. strain Tf were 30$^{\circ}$C and between pH 5.5-7.3, respectively. Aeration also improved greatly growth and yield of biomass of Photorhabdus sp. strain Tf. Photorhabdus sp. strain Tf in batch fermentation showed growth-associated pattem in terms of pigment production, and the pH of culture medium rose steadily until growth stopped dUling the fermentation. Both pigment production and culture pH rise would be useful parameters indicating a reliable growth of Photorhabdus sp. strain Tf.

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Medium Optimization for the Protease Production by Bacillus licheniformis Isolated from Cheongkookjang (청국장에서 분리된 Bacillus licheniformis의 Protease 생산을 위한 배지 최적화)

  • Yoon, Ki-Hong;Shin, Hye-Young
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.385-390
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    • 2010
  • Bacillus licheniformis fermenting soybean product with highest score in consumer acceptance had been isolated from homemade Cheongkookjang. In order to develop the medium composition, effects of ingredients including nitrogen sources, carbon sources, metal ions and phosphate were examined for protease production of the isolate. Potato starch increased the protease productivity, while glucose repressed it. Yeast extract was the most effective nitrogen source for enzyme production. The calcium was found to increase protease activity slightly while cell growth and enzyme production was completely inhibited by divalent ions such as $Zn^{2+}$, $Cu^{2+}$ and $Co^{2+}$. The maximum protease productivity was reached approximately 800 unit/mL in the optimized medium consisting of potato starch (1.5%), yeast extract (1.5%), $CaCl_2$(0.7%), $K_2HPO_4$(0.03%) and $KH_2PO_4$(0.03%). The protease activity of culture filtrate was gradually decreased after incubation for 28 h.