• Title/Summary/Keyword: 배지 최적화

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Production and Characterization of Thermostable Protease from Bacillus licheniformis Isolated from Korean Traditional Soybean Paste (재래식 된장에서 분리된 Bacillus licheniformis의 내열성 Protease 특성과 생산성)

  • Bae, Young Eun;Yoon, Ki-Hong
    • Korean Journal of Microbiology
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    • v.48 no.4
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    • pp.298-304
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    • 2012
  • Among 63 Bacillus strains grown at $60^{\circ}C$ from sixteen samples of homemade Korean soybean paste, one strain was selected for producing the thermostable protease. The isolate has been identified as Bacillus licheniformis on the basis of its 16S rDNA sequence, morphology and biochemical properties. Culture filtrate of the isolate showed maximal protease activity at the reaction condition of $60-65^{\circ}C$ and pH 11. The culture filtrate retained more than 87% of initial protease activity after incubation for 30 min at $60^{\circ}C$ without substrate. 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. Lactose and soytone peptone were the most effective carbon and nitrogen source for the enzyme production. After the late logarithmic growth phase the isolate began to produce the protease, and the maximum protease productivity was reached to 550 unit/ml in the optimized medium consisting of lactose (3%), soytone peptone (1.5%), $MgSO_4$ (0.1%), $K_2HPO_4$ (0.03%), and $KH_2PO_4$ (0.03%) at 28 h of incubation.

Optimal Conditions of Mycelial Growth and Exopolysaccharide Production in Submerged Culture of Phellinus baumii (Phellinus baumii으로부터 세포외 다당체 생산의 최적화)

  • Hwang, Hye-Jin;Kim, Sang-Woo;Yun, Jong-Won;Park, Jang-Won
    • Journal of Life Science
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    • v.14 no.1
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    • pp.51-56
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    • 2004
  • The polysaccharide isolated from Phellinus species has been known as a folk remedy, including antitumor and immune-stimulating activities. However, there are lacks of knowledge about mycelial growth and exopolysaccharide (EH) production in its submerged culture. We investigated the optimal conditions on mycelial growth and EPS production in Phellinus baumii. The optimal temperature and initial pH for mycelial growth and EPS production in shake flask culture of P. baumii were proved to be 3$0^{\circ}C$ and pH 5.0, respectively. In case of carbon source, cellobiose and maltose were highly efficient for mycelial growth and fructose and mannitol were also relatively favorable for EPS production. Yeast extract was the most suitable nitrogen source for mycelial growth and EPS production. The composition of optimal culture medium was determined to be fructose 20 g/L, yeast extract 20 g/L, and $CaCl_2$ 0.55 g/L, respectively. Under the optimal culture condition, the maximum mycelial biomass and EPS achieved in a 5-L stirred-tank fermenter were 17.43 g/L and 3.6 g/L, respectively. It was found that the EPS was a glycoprotein onsisted of mainly arginine (14.1%) and glycine (12.0 %) in protein moiety and mainly mannose (48.7%) and arabinose (38.4%) in carbohydrate moiety.

Antifungal Activities of Pseudomonas spp. Strains Against Plant Pathogens and Optimization of Culture Conditions (식물병원성 진균에 항균 효과를 지닌 슈도모나스 균주의 항진균 활성 증진을 위한 배양조건의 최적화)

  • Chang, Seog-Won;Choi, Byung-Jin;Hong, Jeum-Kyu;Rho, Yong-Taek
    • Korean Journal of Microbiology
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    • v.46 no.3
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    • pp.248-254
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    • 2010
  • To define the optimum conditions for the mass production of four antifungal Pseudomonas spp. isolated from soil, we have investigated culture conditions and effects of various nutrient sources on the bacterial growth and evaluated antagonistic activity against Rhizoctonia solani and Sclerotinia homoeocarpa, plant pathogens. The optimum temperature and pH for the growth of these isolates were determined as pH 7.0 and $20^{\circ}$ or $25^{\circ}C$, respectively. Sucrose, tryptone, and $K_2HPO_4$ generally were more adequate for better growth as carbon, nitrogen and mineral source, respectively. The nutrient sources were also found to be very effective for high antifungal activities against R. solani and S. homoeocarpa. It was elucidated that YUD-F group (P. mandelii and P. fluorescens), which inhabit regions at relatively low temperature, had more broad spectrum and higher antifungal activity than YUD-O group (P. trivialis and P. jessenii) generally against R. solani and S. homoeocarpa. It is thought that the differences of the average temperature in the various habitats of Pseudomonas spp. influence the optimal growth temperature and antifungal activity. Especially, Pseudomonas spp. of YUD-O group showed the better antifungal activity against dollar spot caused by S. homoeocarpa, but showed relatively weaker antifungal activity against brown patch caused by R. solani.

Isolation of a Phenol-degrading Bacterial Strain and Biological Treatment of Wastewater Containing Phenols (Phenol 분해균주의 분리 및 페놀함유 폐수의 생물학적 처리)

  • Lee, Hyun Don;Lee, Myoung Eun;Kim, Hyung Gab;Suh, Hyun-Hyo
    • Journal of Life Science
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    • v.23 no.10
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    • pp.1273-1279
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    • 2013
  • Aromatic hydrocarbons, such as phenol, have been detected frequently in wastewater, soil, and groundwater because of the extensive use of oil products. Bacterial strains (56 isolates) that degraded phenol were isolated from soil and industrial wastewater contaminated with hydrocarbons. GN13, which showed the best cell growth and phenol degradation, was selected for further analysis. The GN13 isolate was identified as Neisseria sp. based on the results of morphological, physiological, and biochemical taxonomic analyses and designated as Neisseria sp. GN13. The optimum temperature and pH for phenol removal of Neisseria sp. GN13 was $32^{\circ}C$ and 7.0, respectively. The highest cell growth occurred after cultivation for 30 hours in a jar fermentor using optimized medium containing 1,000 mg/l of phenol as the sole carbon source. Phenol was not detected after 27 hours of cultivation. Based on the analysis of catechol dioxygenase, it seemed that catechol was degraded through the meta- and ortho-cleavage pathway. Analysis of the biodegradation of phenol by Neisseria sp. GN13 in artificial wastewater containing phenol showed that the removal rate of phenol was 97% during incubation of 30 hours. The removal rate of total organic carbon (TOC) by Neisseria sp. GN13 and activated sludge was 83% and 78%, respectively. The COD removal rate by Neisseria sp. GN13 from petrochemical wastewater was about 1.3 times higher than that of a control containing only activated sludge.

Optimization of Treatment Concentration and Screening of Exogenous Plant Growth Regulators for Improvement of Salidroside Yield in Rhodiola sachalinensis A. Bor Cell Suspension Cultures (참돌꽃 (Rhodiola sachalinensis A. Bor) 현탁세포배양에서 Salidroside 생산을 위한 외재 호르몬의 영향 및 처리 농도의 최적화)

  • Choi Hye-Jin;Kim Su-Jeong;Hwang Baik;Ahn Jun-Cheul
    • Journal of Plant Biotechnology
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    • v.32 no.2
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    • pp.105-109
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    • 2005
  • To enhance salidroside productivity from a cell suspension cultures of Rhodiola sachalinensis, various combinations of auxin (NAA) and cytokinins (BA, Kinetin) concentration and addition of $GA_3$, TDZ, zeatin, spermine and spermidine at a hormone combination to be established were examined. NAA/BA combination is superior to NAA/kinetin combination in biomass and salidroside content. Maximum salidroside production ($64.6{\pm}8.9\;mg/L$ medium) was obtained from $2B_5$ medium with 1 mg/L NAA and 5 mg/L BA. The adding of $GA_3$ ($0.01\;mg/L{\sim}5\;mg/L$) was beneficial for salidroside accumulation and the highest productivity of salidroside, $90.3{\pm}8.34\;mg/L$, was obtained from $2B_5$ medium supplemented with 1 mg/L NAA, 5 mg/L BA and 0.1 mg/L $GA_3$. On TDZ, zeatin, spermine and spermidine, expectant results were not obtained except a little affirmative effect of spermidine ($69{\pm}2.88$ at 1 mg/L).

Optimization of Xylitol Production by Candida tropicalis in Two-stage Fed-batch Culture (Candida tropicalis의 2단계 유가식 배양에 의한 Xylitol 생산의 최적화)

  • 유연우;조영일;서진호
    • KSBB Journal
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    • v.17 no.1
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    • pp.93-99
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    • 2002
  • Two-stage fed-batch culture of Candide tropicalis that was designated primarily to cultivate the cell in the glucose medium (1st stage) and then produced the xylitol from xylose medium (2nd stage) was developed to improve a xylitol yield and productivity. In the growth stage, glucose was automatically supplied to the fermentor by pH-stat mode when the pH was up 5.7, When a feeding medium was added in order to reach the glucose and yeast extract concentrations up to 100 and 40 g/L, respectively, a high cell concentration and a relatively low ethanol concentration were obtained in 18.5 h culture. In the production stage, initial xylose concentration of 150 g/L was the most favorable for obtaining the final xylitol concentration and productivity. The addition of mineral salts was also enhanced a xylitol production. But the aeration rate was not significantly affected a xylitol production. When the addition of 16 g yeast extract and 232.5 g xylose powder at the production stage was used, xylitol yield and productivity were significantly increased. With these conditions, xylitol concentration, yield and productivity of 108.9 g/L, 74%) and 3.3 g/L·h, respectively, were obtained in a final volume of 1.58 L. The further addition of 16 g yeast extract and 232.5 g xylose powder increased the working volume partly (1.67 L) and resulted in a relatively high xylitol concentration, yield and productivity of 193 g/L, 70% and 3.6 g/L·h, respectively.

Improvement of Growth and Benzo[c]phenanthridine Alkaloids Production by Modifying Nitrogen Source in Suspension Cell Culture of Eschscholtzia californica (Eschscholtzia californica의 현탁 세포배양에서 질소원 조절에 의한 세포 성장 및 Benzo[c]phenanthridine Alkaloids 생산량 향상)

  • Lee, Song-Eun;Rhee, Hong-Soon;Son, Seok-Young;Park, Jong-Moon
    • KSBB Journal
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    • v.24 no.2
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    • pp.195-200
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    • 2009
  • The effect of nitrogen source on cell growth and benzo[c]phenanthridine alkaloids production by modifying $NO_3\;^-:NH_4\;^+$ ratio in cell suspension culture of Eschscholtzia califarnica was investigated. When total nitrogen concentration is maintained (60 mM), maximum benzo[c]phenanthridine alkaloids production is about 60.72 mg/L at 50:10 (mol/mol). This productivity was 3.8 times higher than that obtained when cells were grown instandard MS medium. The decrease of $NO_3\;^-:NH_4\;^+$ ratio at 60 mM of total nitrogen caused the decline of both growth and benzo[c]phenanthridine alkaloids production. Under the same concentration of $N0_3\;^-$ (50 mM), higher concentration of $NH_4\;^+$ inhibited cell growth strongly but induced alkaloids production slightly. Also, under the same concentration of $NH_4\;^+$ (25 mM), higher concentration of $N0_3\;^-$ induced alkaloids production strongly but high concentration of $N0_3\;^-$ (${\geq}$100 mM) interfered alkaloids instead. Maximum benzo[c]phenanthridine alkaloids production is about 62.71 mg/L at 50:25 (mol/mol). These results suggest that higher biomass and higher alkaloids production could be obtained by optimizing each nitrogen concentration as well as $NO_3\;^-:NH_4\;^+$ ratio in the culture medium. Nitrate and ammonium in culture medium have distinct role in the regulation of growth and alkaloids production; ammonium had a strong influence on growth while nitrate had an influence on alkaloids production.

Optimization of Submerged Culture Conditions for the Growth Increase of Ginseng Adventitious Root Containing Germanium (게르마늄 함유 인삼 부정근의 생장 증대를 위한 액체배양 조건의 최적화)

  • Chang, Eun-lung;Oh, Roon-II
    • Journal of Ginseng Research
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    • v.33 no.2
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    • pp.143-148
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    • 2009
  • This study was carried out to detennine the optimal submerged culture conditions for the growth increase of ginseng adventitious roots containing germanium by means of a fractional factorial design with four factors and three levels, using the response surface methodology (RSM). The ginseng (Panax ginseng CA. Meyer) adventitious roots were induced by plant growth regulators and cultured in a liquid SH medium. The effects of various $GeO_2$ and phosphoric acid ($H_3P0_4$) concentrations in the medium, $GeO_2$ addition time and the pH of the medium on the fresh weight of the ginseng adventitious roots were investigated. The optimum pH of the medium and the phosphoric acid concentration detennined by the partial differentiation of the model equation were 4.7 and 6.0 roM, respectively. The predicted optimal $GeO_2$ concentration was 10 ppm and the $GeO_2$ addition time did not affect the growth of ginseng adventitious roots. Under these conditions, the growth of the ginseng adventitious root containing germanium was predicted to be 2.47 g.

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.

Screening of Lactic Acid Bacteria for Strong Folate Synthesis and Optimization of Fermentation (고엽산 생산능의 유산균 탐색 및 발효 조건 최적화)

  • Du, Kyung Min;Park, Se Jin;Park, Myung Soo;Ji, Geun Eog
    • Korean Journal of Food Science and Technology
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    • v.46 no.3
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    • pp.328-333
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    • 2014
  • Folate is a water-soluble vitamin B that is required for the synthesis of amino acids and nucleic acids. It plays an important role in cell division and cell growth in several living organisms. The purpose of this study was to screen strong folate-synthesizing bacteria and to optimize their culture conditions for folate production. Folate production was quantified by microbiological assays by using folate-dependent strain Lactobacillus rhamnosus KCTC 3237. Folate derivatives were identified by LC-MS/MS. Of the 65 strains of bifidobacteria and lactobacilli tested, L. plantarum Fol 708 demonstrated the greatest ability to produce folate. Its optimal pH for folate production was 5.5 in a pH-controlled, lab-scale fermenter. Coculturing L. plantarum Fol 708 with L. brevis GABA 100 in a milk medium enhanced the level of folate produced in comparison to culturing L. plantarum Fol 708 alone.