• Title/Summary/Keyword: optimum medium

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Suspension Culture of Gardenia jasminoides Ellis Cell for Production of Yellow Pigment

  • Kim, Sang-Hwa;Park, Young-Goo;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.1 no.2
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    • pp.142-149
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    • 1991
  • Gardenia callus was induced in MS medium containing $10{\;}{\mu}M$ of 2,4 diphenoxy acetic acid (2,4-D), $1{\;}{\mu}M$ kinetin, and 3% sucrose in the dark. $B_5$ medium was identified to be the most adequate medium for cell growth. Indole-3-acetic acid (IAA) was better growth regulator than 2,4-D not only for cell growth but slso for carotenoid production. Ligt also played a critical role on synthesis of carotenoid. Gardenia cells grown in $B_5$ medium could utilize a polysaccharide, soluble starch, as a carbon source. The cell growth was stimulated in $B_5$ medium fortified with 0.2% yeast extract. The optimum pH for cell growth was 5.7. High density cultures can be maintained by increasing inoculum size and medium concentration accordingly. Specific growth rate and mass doubling time were 0.095 $day^{-1}$ and 7.3 days, respectively. The cell immobilized in alginate tends to formulate more enlarged vacuoles containing yellow pigment compared with those of suspended cell. Carotenoid content of immobilized cell was about $264.4{\;}{\mu}g/g$ fresh weight (F.W.) corresponding twice of the content of suspended cell ($112.08{\;}{\mu}g/g$ F.W.). The color of gardenia cell was shifted from yellow to red when carbohydrase-secreting fungus, Trichoderma reesei, was co-cultivated with gardenia cells.

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Culture Optimization for Bacillus lentimorbus G-74 by Using a Miscanthus purpurascens Juice Medium (억새즙액 배지를 이용한 Bacillus lentimorbus G-74 균주의 배양 최적화)

  • Kang, Sun-Chul;Seo, Hae-Jeong
    • Korean Journal of Environmental Agriculture
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    • v.23 no.1
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    • pp.68-74
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    • 2004
  • Miscanthus purpurascens Juice containing potassium (37,952 mg/L), nitrogen (14,000 mg/L), phosphorus (6,800 mg/L), magnesium (5,969 mg/L), calcium (5,910 mg/L), etc., was investigated to develop a novel meidum far the mass cultivation of useful microorganisms. For this research, we first isolated an antagonistic bacterium G-74 from soil, which showed strong growth inhibition against two phytopathogenic fungi, Rhizoctonia solani and Botrytis cinerea, and identified as Bacillus lentimorbus G-74 based on the morphological characteristics and MIDI analysis. Culture conditions for G-74 strain in the M. purpurascens juice medium were optimized. Dilution rate of the medium, temperature and initial pH for the optimum growth of G-74 strain were 30% (V/V), $35^{\circ}C$ and 5.0, respectively. It was found that additions of 2.0% (W/V) corn starch as a carbon source and 1.0% (W/V) yeast extract as a nitrogen source in this medium increased B. lentimorbus G-74 growth to 66% more efficient than Luria Bertani medium.

Effects of GA3 and Charcoal on Plant Regeneration from Somatic Embryos of Acanthopanax sessiliflorus (오가피(Acanthopanax sessiliflorus)의 체세포배로부터 식물체 재생에 미치는 GAa3와 Charcoal의 영향)

  • Lee, Kang-Seop;Choi, Yong-Eui;Sim, Ock-Kyeong;Joo, Sun-Ah;Shin, Jeong-Sun;Jeong, Jae-Hun;Kim, Young-Shin;Kim, Ee-Yup
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.253-257
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    • 2002
  • To establish the optimum condition for plant regeneration from somatic embryos of Acanthopanax sessiliflorus Rupr. et Maxim, a medicinal plant, somatic embryos were induced from zygotic embryo-derived embryogenic callus in hormoen-free MS medium. To induce plantlet conversion, cotyledonary somatic embryos were cultured on MS solid medium with GA$_3$at various concentrations (0~10 mg/L) for three weeks. Plantlets were transferred to 1/3 MS solid medium with 0.5% charcoal for 7 weeks. Stem length was increased proportionally to the concentration and treatment period of GA$_3$. Also, the highest leaf width (8.9 mm) and leaf number (2.84) of plantlet were obtained when plantlets were converted on 5,10 mg/L GA$_3$pretreatments, respectively. The highest plant conversion frequency (66.7%) was obtained when the somatic embryos were cultured on medium containing 5 mg/L GA$_3$ for 3 weeks and then were transferred to 1/3 MS medium with 0.5% charcoal. The highest survival rate of soil transfer was 90% when plantlets were regenerated on medium with 5 mg/L GA$_3$ for 3 weeks and then transferred to plastic pots containing vermiculite and sand mixture for 4 weeks.

Study of the Production of Alkaline Keratinases in Submerged Cultures as an Alternative for Solid Waste Treatment Generated in Leather Technology

  • Cavello, Ivana A.;Chesini, Mariana;Hours, Roque A.;Cavalitto, Sebastian F.
    • Journal of Microbiology and Biotechnology
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    • v.23 no.7
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    • pp.1004-1014
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    • 2013
  • Six nonpathogenic fungal strains isolated from alkaline soils of Buenos Aires Province, Argentina (Acremonium murorum, Aspergillus sidowii, Cladosporium cladosporoides, Neurospora tetrasperma, Purpureocillium lilacinum (formerly Paecilomyces lilacinus), and Westerdikella dispersa) were tested for their ability to produce keratinolytic enzymes. Strains were grown on feather meal agar as well as in solid-state and submerged cultures, using a basal mineral medium and "hair waste" as sole sources of carbon and nitrogen. All the tested fungi grew on feather meal agar, but only three of them were capable of hydrolyzing keratin, producing clear zones. Among these strains, P. lilacinum produced the highest proteolytic and keratinolytic activities, both in solid-state and submerged fermentations. The medium composition and culture conditions for the keratinases production by P. lilacinum were optimized. Addition of glucose (5 g/l) and yeast extract (2.23 g/l) to the basal hair medium increased keratinases production. The optimum temperature and initial pH for the enzyme production were $28^{\circ}C$ and 6.0, respectively. A beneficial effect was observed when the original concentration of four metal ions, present in the basal mineral medium, was reduced up to 1:10. The maximum yield of the enzyme was 15.96 $U_c/ml$ in the optimal hair medium; this value was about 6.5-fold higher than the yield in the basal hair medium. These results suggest that keratinases from P. lilacinum can be useful for biotechnological purposes such as biodegradation (or bioconversion) of hair waste, leading to a reduction of the environmental pollution caused by leather technology with the concomitant production of proteolytic enzymes and protein hydrolyzates.

Optimization of Culture Medium for Novel Cell-Associated Tannase Production from Bacillus massiliensis Using Response Surface Methodology

  • Belur, Prasanna D.;Goud, Rakesh;Goudar, Dinesh C.
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.199-206
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    • 2012
  • Naturally immobilized tannase (tannin acyl hydrolase, E.C. 3.1.1.20) has many advantages, as it avoids the expensive and laborious operation of isolation, purification, and immobilization, plus it is highly stable in adverse pH and temperature. However, in the case of cell-associated enzymes, since the enzyme is associated with the biomass, separation of the pure biomass is necessary. However, tannic acid, a known inducer of tannase, forms insoluble complexes with media proteins, making it difficult to separate pure biomass. Therefore, this study optimizes the production of cell-associated tannase using a "protein-tannin complex" free media. An exploratory study was first conducted in shake-flasks to select the inducer, carbon source, and nitrogen sources. As a result it was found that gallic acid induces tannase synthesis, a tryptose broth gives higher biomass, and lactose supplementation is beneficial. The medium was then optimized using response surface methodology based on the full factorial central composite design in a 3 l bioreactor. A $2^3$ factorial design augmented by 7 axial points (${\alpha}$ = 1.682) and 2 replicates at the center point was implemented in 17 experiments. A mathematical model was also developed to show the effect of each medium component and their interactions on the production of cell-associated tannase. The validity of the proposed model was verified, and the optimized medium was shown to produce maximum cell-associated tannase activity of 9.65 U/l, which is 93.8% higher than the activity in the basal medium, after 12 h at pH 5.0, $30^{\circ}C$. The optimum medium consists of 38 g/l lactose, 50 g/l tryptose, and 2.8 g/l gallic acid.

Effect of Basal Medium and Plant Growth Regulator on in vitro Plant Regeneration from Axillary Buds of Walnut New Cultiver "Sinlyeong"

  • Kwon, Young Hee;Lee, Joung Kwan;Kim, Hee Kyu;Kim, Kyung Ok;Park, Jae Seong;Huh, Yoon Sun
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.10a
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    • pp.15-15
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    • 2019
  • The walnut (Juglans regia L.), a member of the Juglandaceae, is native to the mountain ranges of central Asia. This species of walnut is valued commercially for its nuts and in some areas for its timber. The seeds of walnut are recalcitrant and it has strong integument dormancy and their germination is irregular, making its natural propagation difficult. Low percentage of seed germination and long propagation cycle are the main problems of propagation. This study was conducted medium composition on in vitro plantlet regeneration from axillary buds of walnut. It has proved to be the most generally applicable and reliable method of in vitro propagation. Micropropagation culture that axillary buds are excised aseptically enables faster multiplication of plants. The axillary buds of walnut new cultivar "Sinlyeong" were cultured on two basal media which contained the different plant growth regulators depending on the respective shooting and rooting stage. After 12 weeks, the shoot generation rate was 85.3%, the shoot number and its length were 1.9/explant and 2.7 cm in the most favorable medium composition. The percentage of rooting was 25.4%. From these results, it was found the optimum basal medium and plant growth regulator for in vitro plant regeneration from axillary buds of walnut new cultivar "Sinlyeong". However, we have continued to search the other medium additives to enhance the rate of walnut root.

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Studies on the Hydrolysis of Inulin in Jerusalem Artichokes by Fungal Inulase (미생물(微生物) Inulase에 의(依)한 돼지감자 중의 Inulin분해(分解)에 관한 연구(硏究))

  • Kim, Ki-Choul
    • Applied Biological Chemistry
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    • v.18 no.3
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    • pp.177-182
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    • 1975
  • The analysis of Jerusalem artichoke showed that it contains 12.09% of Inulin. The results obtained from the examination of the conditions for fructose production by cultivating Pencillum sp 1 in the Jerusalem articoke medium were as follows: 1. The optimum amount of water added to Jerusalem artichoke was 2.5 $\ell$ of distilled water per ㎏ of fresh Jerusalem artichoke. It this case, the concentration of Inulin was 4% (w/v). 2. The optimum temperature was $30^{\circ}C$, the initial optimum pH was 5.0 and the optimum cultural period was 72 hours. 3. Shaking culture with 50 ml of the medium and 120 oscills/min in 500 ml shaking flask was most effective as the culture method. 4. 0.1% of $NH_4H_2PO_4$ as a nitrogen source, 0.001 of $FeSO_47H_20$ and 0.001% of $MgSO_47H_2$ as metal salts were most effective. 5. Fructose production continued to increase for 72 hours under the optimum conditions for cultivation and the highest production rate to the Inulin was 95.25%.

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Optimum Seeding Rate in Different to Soil Salinity for Broadcasting on the Rice Flooded Paddy Surface at South-western Reclaimed Saline Land of Korea (서남부 간척지에서 벼 담수표면산파재배시 토양 염농도별 적정 파종량)

  • Back, Nam-Hyun;Choi, Weon-Young;Ko, Jong-Cheol;Park, Hong-Kyu;Nam, Jeong-Kweon;Park, Kwang-Geun;Kim, Sang-Su;Kim, Bo-Kyeong;Kim, Choung-Kon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.spc1
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    • pp.47-51
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    • 2006
  • This study was conducted to establish the optimum seeding rate in different soil salinity level for yield stability of broadcasting on flooded paddy surface to the reclaimed saline land of south-western part at Gyehwado substation of the Honam Agricultural Research institute in $2003{\sim}2004$. Soeganbyeo was tested in the Munpo series (fine sand loam) the results obtained is as follows: As seeding rate was higher, the number of seeding stand was increased and the number of seeding stands in the low salinity field is sharply increased than those of the medium salinity field. The length of culm in medium salinity field tends to be shorter than that of the low salinity field and as seeding rate was increased, the lodging is severe. The milled rice yield was increased as up to 9 kg/10a in low and medium salinity soil. Complete rice was no significantly increased over 5 kg/10a seeding rate in low salinity field and over 7 kg/10a seeding rate in medium salinity field. Considering the yield of milled and complete rice, seeding stand and lodging, The proper seeding rate is $5{\sim}7kg/10a$ in low salinity and $7{\sim}9kg/10a$ in medium salinity for broadcasting on flooded paddy surface at the reclaimed saline land of southwestern part.

Optimization of Medium Components for the Production of Crude Biosurfactant by Bacillus subtilis JK-1 (Bacillus subtilis JK-1의 생물계면활성도를 위한 최적 배지 조성)

  • Joo, Myeong-Hoon;Kim, Ji-Yeon
    • Journal of Applied Biological Chemistry
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    • v.54 no.1
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    • pp.7-14
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    • 2011
  • Bacillus subtilis JK-1 showed degradation activity against crude oil, gasoline, kerosene, and light oil, and this strain was used as a crude biosurfactant producing microorganism in this study. To optimize the culture medium for production of crude biosurfactant, the influences of various carbon, nitrogen and mineral sources were assessed. The highest biosurfactant production by B. subtilis JK-1 was observed after 96 h cultivation, containing 1.0% (w/v) soluble starch as a carbon source and 0.5% (w/v) skim milk as a nitrogen source, and carbon to nitrogen concentraion (C/N) ratio was 2.0. For the biosurfactant production 0.1% (w/v) of $KNO_3$ was the most effective mineral source. Comparison of biosurfactant production indicates that B. subtilis JK-1 produces more biosurfactant in the optimum medium established in this study than LB and TSB. Under the optimum medium, the surface tension of culture broth of B. subtilis JK-1 was decreased from 47.3 dyne/cm to 24.0 dyne/cm after cultivation of 48 h.

Production and Structural Analysis of Cellulose by Acetobacter sp. V6 Using Static Culture (정치배양을 이용하여 Acetobacter sp. V6의 셀룰로오스 생산 최적화 및 구조 분석)

  • Kim, Jeong-Do;Jung, Ho-Il;Jeong, Jin-Ha;Park, Ki-Hyun;Jeon, Young-Dong;Hwang, Dae-Youn;Lee, Chung-Yeol;Son, Hong-Joo
    • Korean Journal of Microbiology
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    • v.45 no.3
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    • pp.275-280
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    • 2009
  • The optimal medium compositions for the production of bacterial cellulose (BC) by a Acetobacter sp. V6, which was isolated from the traditionally fermented vinegar in Korea, were investigated in static cultures. The optimum medium compositions for BC production were 3% glucose, 3% soytone, 0.8% $K_2HPO_4$, and 0.4% ethanol, respectively. Adding $NaH_2PO_4$ or $KH_2PO_4$ had not shown the increase in BC production. Under the optimum medium compositions, the highest BC production was 44.67 g/$m^2$ in 8 days and the thickness of BC pellicle was about 1 cm. Structural properties of BC produced in the optimal medium were studied using Fourier-transform infrared spectroscopy and X-ray diffractometer. BC from the optimal medium was found to be of cellulose type I, the same as typical native cellulose. No difference in the compositions between bacterial and plant celluloses, but BC showed unique micro-network structure and high crystallinity (82%).