• Title/Summary/Keyword: optimization of culture conditions

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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.

Strain Selection and Optimization of Mixed Culture Conditions for Lactobacillus pentosus K1-23 with Antibacterial Activity and Aureobasidium pullulans NRRL 58012 Producing Immune-Enhancing β-Glucan

  • Sekar, Ashokkumar;Kim, Myoungjin;Jeong, Hyeong Chul;Kim, Keun
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.697-706
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    • 2018
  • Lactobacillus pentosus K1-23 was selected from among 25 lactic acid bacterial strains owing to its high inhibitory activity against several pathogenic bacteria, including Escherichia coli, Salmonella typhimurium, S. gallinarum, Staphylococcus aureus, Pseudomonas aeruginosa, Clostridium perfringens, and Listeria monocytogenes. Additionally, among 13 strains of Aureobasidium spp., A. pullulans NRRL 58012 was shown to produce the highest amount of ${\beta}$-glucan ($15.45{\pm}0.07%$) and was selected. Next, the optimal conditions for a solid-phase mixed culture with these two different microorganisms (one bacterium and one yeast) were determined. The optimal inoculum sizes for L. pentosus and A. pullulans were 1% and 5%, respectively. The appropriate inoculation time for L. pentosus K1-23 was 3 days after the inoculation of A. pullulans to initiate fermentation. The addition of 0.5% corn steep powder and 0.1% $FeSO_4$ to the basal medium resulted in the increased production of lactic acid bacterial cells and ${\beta}$-glucan. The following optimal conditions for solid-phase mixed culture were also statistically determined by using the response surface method: $37.84^{\circ}C$, pH 5.25, moisture content of 60.82%, and culture time of 6.08 days for L. pentosus; and $24.11^{\circ}C$, pH 5.65, moisture content of 60.08%, and culture time of 5.71 days for A. pullulans. Using the predicted optimal conditions, the experimental production values of L. pentosus cells and ${\beta}$-glucan were $3.15{\pm}0.10{\times}10^8CFU/g$ and $13.41{\pm}0.04%$, respectively. This mixed culture may function as a highly efficient antibiotic substitute based on the combined action of its anti-pathogenic bacterial and immune-enhancing activities.

Optimization of submerged culture conditions for the mycelial growth and exo-biopolymer production by Cordyceps millitaris

  • Park, Jong-Pil;Sinha, Jayanta;Song, Chi-Hyeon;Yun, Jong-Won
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.317-320
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    • 2000
  • The optimal temperature and pH for both mycelial growth and exe-biopolymer production by Cordyceps millitaris in shake flask culture were found to be $20^{\circ}C$ and 6.0, respectively. Sucrose (4%) and corn steep powder (1%) were the most suitable carbon and nitrogen source for mycelial growth and exo-biopolymer production. The maximum specific growth rate $(0.142h^{-1})$ was achieved when sucrose was used as the sole carbon source. Exo-biopolymer production was increased with the increase in C/N molar ratio concentration, probably due to the facilitated carbon uptake. Under the optimal culture conditions, the maximum mycelial growth exe-biopolymer concentration were reached to around 13.3 g dry cell weigh/l and 3.33 g/l, respectively.

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Characteristics of Culture Conditions for the Production of Crude Biosurfactant by Bacillus subtilis JK-1 (Bacillus subtilis JK-1의 생물계면활성제 생산을 위한 배양 특성)

  • Kim, Ji-Yeon
    • Journal of Applied Biological Chemistry
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    • v.54 no.3
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    • pp.153-158
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    • 2011
  • Optimal culture conditions were characterized for production of crude biosurfactant of Bacillus subtilis JK-1. During incubation of B. subtilis JK-1, the bacterial growth pattern, changes of the surface tension at variable temperatures, pH and NaCl concentrations in bacterial culture medium were studied. The strain was able to grow and produce biosurfactant at $15-45^{\circ}C$, in the pH range of 6-10, and at 0-10% (w/v) NaCl. In case, culture broth pH was gradually changed to neutral or weak alkaline. Optimal culture conditions for crude biosurfactant production were at $35^{\circ}C$ and pH 7.0 after 48 h incubation and the surface tension of biosurfactant was 24.0 mN/m. Besides, as the concentration of NaCl was increased from 0 to 10% (w/v), the growth was decreased, pH of the culture broth was converted from weak alkaline to acidic, and the surface tension rised.

Optimization of Culture Conditions of Chitosanase-producing Bacillus sp. P16 (키토산분해효소 생산을 위한 Bacillus sp. P16 배양조건의 최적화)

  • Jung, Mi-Ra;Jo, Yu-Young;Chil, Youn-Tae;Park, Ro-Dong
    • Applied Biological Chemistry
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    • v.42 no.3
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    • pp.193-198
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    • 1999
  • The optimal culture condition of Bacillus sp. P16 was investigated for production of an extracellular endo-splitting chitosanase. The best carbon and nitrogen sources for the chitosanase production were chitosan and tryptone, respectively. The best condition for the maximum activity was at $37^{\circ}C$ in a medium containing 0.5% powdered chitosan, 1% tryptone, and 1% NaCl(at initial pH 7.0) in a rotary shaker(200 rpm). In a jar fermenter, the culture duration shortened to $6{\sim}12$ hr for maximum activity and the enzyme activity increased about 100% compared with that of flask culture.

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Statistical Optimization of Culture Conditions for the Production of Aphicidal Metabolites of Beauveria bassiana Bb08 (Beauveria bassiana Bb08의 살충성 물질 생산을 위한 배양조건의 통계적 최적화)

  • Go, Eunsu;Lim, Younghoon;Jeong, Hyeongchul;Choi, Jaepil;Park, Inseo;Kim, Jeong Jun;Lee, Dong-Jin;Kim, Keun
    • Microbiology and Biotechnology Letters
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    • v.41 no.4
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    • pp.398-406
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    • 2013
  • For the maximal production of aphicidal metabolites produced by the Beauveria bassiana Bb08, statistical methods such as the Box-Behnken experimental design and response surface methodology were used. The fungal culture filtrate was sprayed towards 3-star aphids and the mortality was examined. After the statistical analysis of the aphid mortality, the optimal culture conditions were found to be a culture temperature of $26.2^{\circ}C$, medium pH 5.9, flask shaking speed of 209.0 rpm, and culture time of 5.9 days. The expected mortality on days 4, 5, and 6 after spraying the filtrate on to the aphids were 76.8%, 84.9%, and 89.4%, respectively. All 4 factors of the culture conditions significantly affected the production of the aphicidal metabolites, and the order of significance was temperature, pH, culture time and shaking speed.

Optimization of Sweet Rice Muffin Processing Prepared with Oak Mushroom (Lentinus edodes) Powder (표고버섯 첨가 찹쌀머핀의 최적화 및 품질특성)

  • Kim, Bo-Ram;Joo, Na-Mi
    • Journal of the Korean Society of Food Culture
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    • v.27 no.2
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    • pp.202-210
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    • 2012
  • The purpose of this study was to bake sweet rice muffins with oak mushroom ($Lentinus$ $edodes$) powder. The process included substituting sweet rice flour for cake flour and adding oak mushroom powder. This study determined the optimal mixing conditions of oak mushroom muffins by adjusting the amounts of oak mushroom powder, whole eggs, and soybean oil. The mixing conditions for the oak mushroom muffins included 3 categories: oak mushroom powder (X1), whole eggs (X2), and soybean oil (X3) by Central Composite Design (CCD) which was optimized by Response Surface Methodology (RSM). Oak mushroom muffin formulation was optimized using rheology. Yellowness (p<0.001) and redness (p<0.05) displayed a linear model pattern, whereas lightness (p<0.05) was represented by a quadratic model. Among the sensory properties, variables that appeared to show significant values such as appearance (p<0.5), texture (p<0.5), and overall quality (p<0.5) were used to identify optimums. The result of mechanical properties showed significant values in gumminess (p<0.5) and chewiness (p<0.5). The numerical and graphical methods used in this study determined that the optimum formulation for oak mushroom powder sweet rice muffins was 8.75 g of oak mushroom powder, 235.95 g of whole eggs, and 19.93 g of soybean oil.

Production of Alkaline Protease by Entrapped Bacillus licheniformis Cells in Repeated Batch Process

  • Mashhadi-Karim, Mohammad;Azin, Mehrdad;Gargari, Seyyed Latif Mousavi
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1250-1256
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    • 2011
  • In this study, Bacillus licheniformis cells were immobilized by entrapment in calcium alginate beads and were used for production of alkaline protease by repeated batch process. In order to increase the stability of the beads, the immobilization procedure was optimized by statistical full factorial method, by which three factors including alginate type, calcium chloride concentration, and agitation speed were studied. Optimization of the enzyme production medium, by the Taguchi method, was also studied. The obtained results showed that optimization of the cell immobilization procedure and medium constituents significantly enhanced the production of alkaline protease. In comparison with the free-cell culture in pre-optimized medium, about 7.3-fold higher productivity was resulted after optimization of the overall procedure. Repeated batch mode of operation, using optimized conditions, resulted in continuous production of the alkaline protease for 13 batches in 19 days.