• Title/Summary/Keyword: culture pH

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Optimal Production and Characterization of Laccase from Fomitella fraxinea Mycelia (Fomitella fraxinea 균사체로부터 Laccase의 최적생산 및 효소적 특성)

  • Park Kyung-Mi;Park Sang-Shin
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.228-234
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    • 2006
  • The culture conditions were investigated to maximize the production of laccase from Fomitella fraxinea mycelia. Among the tested media, mushroom complete medium (MCM) showed the highest production of the enzyme. The optimum culture medium was 2% dextrose, 0.4% $(NH_4)_{2}HPO_4$, 0.05% $Na_{2}HPO_{4}{\cdot}7H_{2}O$, and 0.05% KCl as carbon, nitrogen, phosphorus, and inorganic salt sources respectively. SDS-PAGE followed by laccase activity staining using 2,6-djmethoxyphenol as the substrate was performed to identify the laccase activity under culture conditions studied. Zymogram analysis of the culture supernatant showed a laccase band with a molecular mass of 50 kDa. The enzyme production from F. fraxinea was reached to the highest level after the cultivation for 10 days at $25^{\circ}C$ and initial pH 8. The enzyme activity of the culture supernatant was most active at $50^{\circ}C$ and pH 5.

Improvement of Ergone Production from Mycelial Culture of Polyporus umbellatus

  • Lee, Wi-Young;Park, Young-Ki;Ahn, Jin-Kwon
    • Mycobiology
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    • v.35 no.2
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    • pp.82-86
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    • 2007
  • Ergone, a fungal metabolite derived from ergosterol, was previously isolated and identified from Polyporus umbellatus. Ergone is a major component of P. umbellatus known to have anti-aldosteronic diuretic effect and also displays cytotoxic activities. Most of mushroom's fruit bodies used for test contained less than 10 ${\mu}g/g$ of ergone. But P. umbellatus have larger amount of ergone than any other mushrooms. In order to improve the ergone production from the submerged culture of P. umbellatus, several factors including medium composition, culture conditions (temperature and pH) and different combinations of co-cultivation with various mycelia were studied. Among various carbon sources examined, starch proved to be most effective for the production of mycelia. The optimum pH and temperature for a flask culture of P. umbellatus mycelia were found to be 4.5 and $25^{\circ}C$, respectively. Under the optimized culture conditions, both the ergone production (86.9 ${\mu}g/g$) and mycelial growth (3.5 g/l) increased when P. umbellatus was cultured with Armillariella mellea. When the optimized conditions were applied, both mycelium and ergone production were significantly enhanced.

Effects of pH on the Elaboration of Pullulan and the Morphology of Aureobasidium pullulans (배양액의 pH가 Aureobasidium pullulans의 풀루란 생성과 형태에 미치는 영향)

  • Shin, Yong-Chul;Byun, Si-Myung
    • Microbiology and Biotechnology Letters
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    • v.19 no.2
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    • pp.193-199
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    • 1991
  • The effects of pH on the cell growth, the elaboration of pullulan, and the morphology of Aureobasidium pullulans IFO 4464 were examined. A. pulluans grew in yeast-like form at constant pH7.5 and in mycelial form at constant pH2.5. At the both pH conditions, the elaboration of pullulan was very low, about 6.0~6.5g/l. The mixture of yeast-like form and mycelial form of cells was found at the constant pH4.5, at which condition, the elaboration of pullulan was high, about 24.5g/l. The pH shift experimemts showed that the specific production rates of pullulan were 0.048($hr^{-1}$)for the mycelial form and 0.058($hr^{-1}$)for the yeast like form, which indicated that the yeast-like form has the similar, only slightly higher, biosynthetic activity of pullulan to the mycelial form at pH4.5 and the pH of culture broth is more important factor for the elaboration of pullulan than the morphology of A. pullulans.

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Production and Characterization of Manganese Peroxidase from the White Rot Fungus Pleurotus ostreatus in Liquid Culture (액체배양한 느타리 버섯균(Pleurotus ostreatus)으로부터 망간퍼옥시데이즈의 생산 및 특성)

  • Lee, Jae-Sung;Ha, Hyo-Cheol
    • Applied Biological Chemistry
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    • v.47 no.1
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    • pp.22-26
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    • 2004
  • The ligninolytic basidiomycete, Pleurotus ostreatus K-2946, was produced a manganese peroxidase (MnP) activity when grown in liquid culture with glucose-yeast-peptone (G-Y-P) medium. However, lignin peroxidase (LiP) was not detected in this culture medium. The purification progress of MnP was purified that included chromatography on Sepharose CL-6B, Superdex 75 prep grade and Mono-Q. MnP purified by column chromatography, was 36400 dalton and a pI of 3.95. The optimal pH and temperature of the purified MnP activity were 5.0 and $55^{\circ}C$. The characteristics of MnP produced was quite similar to those of MnP 3 isoenzyme produced by other strains of P. ostreatus.

Changes in activities of protease, phenoloxidase and cellulase during mycelium growth of Pleurotus ostreatus in sawdust cultures (톱밥배양한 느타리버섯 균사생장시 생산되는 각종 효소변화)

  • Chang, Hyun-You;Kim, Gwang-Po;Cha, Dong-Yeul
    • The Korean Journal of Mycology
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    • v.24 no.2 s.77
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    • pp.149-154
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    • 1996
  • Effects of various kinds of sawdusts, supplements and culture conditions on activities of several enzymes such as protease, phenoloxidase and cellulase produced from mycelium of P. ostreatus grown on sawdust medium were studied and the results are as follows; Higher specific activity of these enzymes was observed when oak tree sawdust and poplar tree sawdust were supplemented with rice bran or wheat bran at rate of 30%, 20% and 10% in total volume respectively. Higher total activities of protease, phenoloxidase and cellulase were observed at 70% of the moisture contents of culture media, while lower activity of these enzymes was observed with 40% moisture contents of sawdust culture medium. The pH 4 and 9 of the sawdust media appeared to be optimum pH for the. production of protease while pH 5 and 7 were optimal for the production of phenoloxidase. The pH 6 of the sawdust medium was optimal for the production of cellulase. The optimum incubating temperature for the production of protease, phenoloxidase and cellulase was $25^{\circ}C$. Higher total activities of protease and phenoloxidase were observed when culture medium was added with wood vinegar at the control, and 0.5% for cellulase.

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Methodology of Genetic Engineering (遺傳工學의 方法論)

  • 이평우;윤경하;이세영
    • Korean Journal of Microbiology
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    • v.13 no.2
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    • pp.71-80
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    • 1975
  • The neurotoxin of Clostridium botulinum type B was purified from a liquid culture. The purification steps consist of ammonium sulfate precipitation of whole culture, treatment of Polymin P(0.15%, v/v), gel filtration on Sephadex G-100 at pH5.6 and DEAE-Sephadex charomatography at pH8.0. The procedure recovered 17% of the toxin assayed in the starting culture. The toxin was homogeneous by sodium dodecyl sulfate(SDS)-polyacrylamide gel electrophoresis and had a molecular weight of 163,000. Subunits of 106,000 and 56,000 molecular weight were found when purified toxin was treated with a disulfide-reducing agent and electro phoresed on SDS-polyacrylamide gels.

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The Effects of Mixed Culture with Aspergillus flavus, Aspergilus niger and Penicillium griseofulvum on Aflatoxin and Patulin Production (Aspeygillus flavus, Aspergillus nicer 및 Peniciilum griseofulvum의 혼합배양이 aflatoxin 및 patulin 생성에 미치는 영향)

  • 강성조;강진순;정덕화
    • Journal of Food Hygiene and Safety
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    • v.16 no.3
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    • pp.206-211
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    • 2001
  • This experiment was conducted to investigate the effects of mixed culture with mycotoxigenic and non-mycotoxigenic fungi on mycotoxin production. For this work, Aspegillus flavus (aflatoxin producing strain), Aspegillus niger (non-mycotoxigenic strain) and Penicillium griseofulvum (patulin producing strain)were cultured in 5 ml SLS medium for 15 days under single or mixed culture. Aflatoxin was determined by direct competitive ELISA, whereas, patulin was measured by HPLC. The mycelial growth, pH and total acidity were also observed by general methods. The mycelial growth was slightly decreased in the mixed culture, meanwhile total acidity was increased and pH was shown lower than that in single culture. Aspergillus flavus produced 145 $\mu\textrm{g}$/ml of aflatoxin for 12 days single culture, but in mixed culture, aflatoxin was decreased to 93%, and was shown as 10.16$\mu\textrm{g}$/ml level. Patulin production in mixed culture was also decreased to 69.3% and was shown only 23.72$\mu\textrm{g}$/ml level as compared with in single culture.

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P(3HB) Accumulation in Alcaligenes eutrophus H16(ATCC 17699) under Nutrient-Rich Condition and Its Induced Production from Saccharides and Their Derivatives

  • Song, Jae-Jun;Shin, Yong-Chul
    • Journal of Microbiology and Biotechnology
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    • v.3 no.2
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    • pp.115-122
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    • 1993
  • Poly(3-hydroxybutyrate)(P(3HB)) accumulation under nutrient-rich condition with various amounts of $(NH_4)_2 SO_4$ was systematically investigated. The results of the electron-microscopy and the solvent extraction showed that the P(3HB) accumulation is unavoidable even under nutrient-rich condition. This indicates that in a two-step culture of Alcaligenes eutrophus H16, the researches should be careful in interpreting the data of polyhydroxyalkanoates(PHAs) accumulation in terms of the carbon-source fed in the second step because the two-step culture product contains the P(3HB) produced under nutrient-rich condition. The polyester production capability in a two-step batch culture of A. eutrophus H16(ATCC 17699) was also investigated using various saccharides and their derivatives such as glucose, fructose, gluconic acid, glucaric acid, sorbitol, lactose, galactose, and mannose. The polyesters synthesized were characterized by 500 MHz$^{1}H-NMR$ spectroscopy, intrinsic viscosity$[\eta]$ measurement in chloroform and differential scanning calorimetry(DSC). 500 MHz $^{1}H-NMR$ analysis showed that all polyesters synthesized generally contained 1~2 mol% of 3HV. Another finding is that the glucose utilization can be increased by changing the autoclaving procedure of the substrate to enhance the P(3HB) production yield up to 46 wt% of P(3HB) in dry cells.

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Evaluation of Metal Biosorption Efficiency of Laboratory-grown Microcystis under Various Environmental Conditions

  • Pradhan, Subhashree;Singh, Sarita;Rai, Lal Chand;Parker, Dorothy L.
    • Journal of Microbiology and Biotechnology
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    • v.8 no.1
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    • pp.53-60
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    • 1998
  • This study examines the effect of pH, temperature, metal ion concentration and culture density on metal biosorption by the nuisance cyanobacterium Microcystis aeruginosa. Ni biosorption was higher at pH 9.2 than at neutral and acidic pH. In contrast the biosorption of Cu and Zn was maximum at pH 7.0. However, biosorption of Zn was difficult to measure at pH values 9.2 and 10.5, owing to the formation of insoluble complexes. All the test metals (Cu, Zn, and Ni) showed maximum biosorption rate at low culture densities of 40 mg dry wt $1^{-1}$. The biosorption of Cu, Zn, and Ni was maximum at $40^{\circ}C$. However, no worthwhile difference in Zn and Ni sorption was noticed at 4 and $29^{\circ}C$ as compared to $40^{\circ}C$. Of these three metals used Microcystis showed a greater binding capacity ($K_{f}$ value=0.84, Freundlich adsorbent capacity) and accelerated biosorption rate for Cu under various environmental conditions. Fitness of mathematical models on metal biosorption by Microcystis confirmed that the biological materials behave in the same way as physical materials. These results suggest that before using a biosorbent for metal recovery, the environmental requirements of the biosorbent must be ascertained.

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Estimation of Residual Biomass, PHB, and Nutrient Concentrations by Supplied Amount of Ammonia Solution in Fermentation of Alcaligenes latus

  • Lee, Yong-Woo;Tsuneo Yamane
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.554-561
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    • 1999
  • A novel estimation method was investigated for determining the concentrations of residual biomass, poly-3-hydroxybutyrate (PHB), and main nutrients including carbon and nitrogen sources, phosphate, and mineral ions from the supplied amount of ammonia solution used for a pH-control solution and nitrogen source in a PHB fermentation. The estimation equations for a batch culture and a fed-batch culture were derived from the relationship between the growth rate of residual biomass and the feed rate of the pH-control solution, and then were applied to the batch culture and the fed-batch cultures of Alcaligenes latus. This method was successfully applied to estimate the concentrations of residual biomass, PHB, and nutrients.

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