• Title/Summary/Keyword: media composition optimization

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Cloning, Characterization of Pichia etchellsii $\beta-Glucosidase$ II and Effect of Media Composition and Feeding Strategy on its Production in a Bioreactor

  • Sethi Benu;Jain Monika;Chowdhary Manish;Soni Yogesh;Bhatia Yukti;Sahai Vikram;Mishra Saroj
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.1
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    • pp.43-51
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    • 2002
  • The cloning and expression of $\beta-glucosidase$ II, encoded by the gene ${\beta}glu2$, from thermotolerant yeast Pichia etchellsii into Escherichia coli is described. Cloning of the 7.3 kb BamHI/SalI yeast insert containing ${\beta}glu2$ in pUC18, which allowed for reverse orientation of the insert, resulted in better enzyme expression. Transformation of this plasmid into E. coli JM109 resulted in accumulation of the enzyme in periplasmic space. At $50^{\circ}C$, the highest hydrolytic activity of 1686 IU/g protein was obtained on sophorose. Batch and fed-batch techniques were employed for enzyme production in a 14 L bioreactor. Exponential feeding rates were determined from mass balance equations and these were employed to control specific growth rate and in turn maximize cell growth and enzyme production. Media optimization coupled with this strategy resulted in increased enzyme units of 1.2 kU/L at a stabilized growth rate of $0.14\;h^{-l}$. Increased enzyme production in bioreactor was accompanied by formation of inclusion bodies.

The Visualization Module Composition for Interface User Platform (인터페이스 사용자 플랫폼의 구현을 위한 시각화 모듈 구성)

  • Kim, Mi-Yun;Seo, Dong-Jo
    • Journal of Korea Multimedia Society
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    • v.16 no.12
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    • pp.1482-1494
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    • 2013
  • As the result of this research, the user platform, which reflects the latest interface technology, supports the ideal interface environment in a future city through using the data from social media. The goal of this research is constructing the user interface platform to approach to the data in an easier and practical condition of the interface environment smart space of the future intelligent city for user. It constructs the main module to organize the platform. Interface user platform modules, which were all based on visualization of service, are constructed by visualization agents, visualization social networks, visualization icons, visualization services, visual data, visual maps and visual classes. In this research, by the variety of services including delivering information, to find the interface environment for the proper interface technology, this research suggests "Interface User Platform" as the result.

In Vitro Development of Porcine Parthenogenetic Embryos under the Oil-free Culture System

  • Park, Sang-Kyu;Choi, Young-Ju;Roh, Sang-Ho
    • Journal of Embryo Transfer
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    • v.25 no.4
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    • pp.259-262
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    • 2010
  • Optimization of the preimplantation mammalian embryo culture condition was widely focused on refining medium composition under the name of chemically defined media. However, recent research revealed that the alteration of physical environment can be a crucial factor to a successful embryo development. In this study, under the same embryo density, a novel culture device named oil-free micro tube culture (MTC) system was evaluated using porcine parthenogenetic embryos. The activated oocytes were placed into the 0.2 ml thin-wall flat cap PCR tube and cultured to the blastocyst stage. As a preliminary step, embryo density and culture medium volume were optimized under a standard drop culture system. The optimal embryo density range for in vitro culture was 0.5 embryos per ${\mu}l$ in $20\;{\mu}l$ drop (20.5%) and 1.0 embryos per ${\mu}l$ in $10\;{\mu}l$ drop (20.6%). Based on these results, we compared drop culture system and 'MTC' system in terms of the developmental rate to the blastocyst stage. In $20\;{\mu}l$ medium volume, the 'MTC' system showed similar blastocyst formation rate when compared with drop culture system (20.2% versus 20.5%, respectively) while the 'MTC' system showed lower blastocyst formation rate than drop culture system in $10\;{\mu}l$ one (12.7% versus 20.0%, respectively). Therefore the $20\;{\mu}l$ MTC system may be an alternative incubation system for short-distance embryo transport without carrying the $CO_2$ incubator and this provides novel embryo culture device to clinical veterinary embryologists.

Optimization of Sawdust Media Composition and Culture Conditions for the Mycelial Growth and Primordia Formation of Agrocybe cylindracea (버들송이(Agrocybe cylindracea)의 균사생장 및 원기형성을 위한 톱밥 배지조성(培地組成) 및 배양조건(培養條件)의 최적화(最適化))

  • Park, Shin;Lee, Jae-Sung
    • The Korean Journal of Mycology
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    • v.18 no.4
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    • pp.198-202
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    • 1990
  • Some factors effecting on the mycelial growth and primordia formation of Agrocybe cylindracea were investigated. Among supplements added into sawdust, wheat bran was the best for the mycelial growth, and its appropriate concentration was 30%. A variety of sawdust preparations was tested singly and in combinations. Oak sawdust and the combination of oak and larch sawdust were the most effective for the mycelial growth. The optimal moisture content was 65%, and the range of optimal temperature was $25\:{\sim}\:28^{\circ}C$. The favorable conditions for the primordia formation appeared to be similar to those of the mycelial growth. Among supplements added into sawdust, wheat bran 30% and the combination of wheat bran and rice bran 30% were the best for the primordia formation. Oak sawdust, and the oak plus larch sawdust and oak plus poplar sawdust were the best for the primordia formation. The optimal temperature for the primordia formation was $17{\pm}2^{\circ}C$.

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Optimization of Monochamus alternatus media and culture period for cordycepin production in Cordyceps militaris culture using solid-state fermentation

  • Ha, Si Young;Jung, Ji Young;Yang, Jae-Kyung
    • Journal of Mushroom
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    • v.19 no.3
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    • pp.126-133
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    • 2021
  • In this study, we investigated the effect of solid culture medium on the production of cordycepin in Cordyceps militaris. The regression equation was expressed as follows: Y1 = 755.3-58.6625X1+4.79432E-14X2-46.6625X3-5.66269E-14X1X2-0.025X1X3+1.62475E-14X2X3-160.6625X12+0.0125X22-206.9625X32, where, Y represents the value of cordycepin content (㎍/g), X1 corresponds to the weight of M. alternatus in solid culture medium (g/bottle), X2 to the water content of the solid culture medium (%), and X3 to the culture period (day). The solid culture medium was optimized using the response surface methodology, and the optimal medium composition was as follows: the weight of M. alternatus in solid culture medium and water content were 16.2% and 100.7% (20.14 mL water/20 g solid culture medium), respectively, with a culture period of 39 days. Under these conditions, the cordycepin content of the fruiting bodies reached 150.0 ㎍/g (actual value). The supplementation of M. alternatus in solid culture for improved cordycepin content of C. militaris seems to be a promising alternative to wild and solid cultivation.

Optimization of Endoglucanase Production from Fomitopsis pinicola Mycelia (Fomitopsis pinicola 균사체로부터 Endoglucanase의 최적생산)

  • Gu, Ji-Min;Park, Sang-Shin
    • Microbiology and Biotechnology Letters
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    • v.41 no.2
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    • pp.145-152
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    • 2013
  • The culture conditions to maximize the production of endoglucanase (EC 3.2.1.4) from the brown rot fungus Fomitopsis pinicola MKACC 54347 mycelia were investigated. Among the tested media for endoglucanase production, Mandel's mineral salts medium (MSM; 1% cellulose, 0.1% peptone, 0.14% $(NH_4)_2SO_4$, 0.03% urea, 0.2% $KH_2PO_4$, 0.03% $MgSO_4{\cdot}7H_2O$, 0.03% $CaCl_2$, and 0.1% trace metal solution (19.8 mM $FeSO_4$, 13.0 mM $MnSO_4$, 12.2 mM $ZnSO_4$, and 15.4 mM $CoCl_2$)) produced the highest activity of the enzyme. To optimize the medium composition for enzyme activity, the effects of various carbon, nitrogen, phosphorus, and inorganic sources were investigated in MSM. Maximal enzyme production was accomplished using a medium containing 2% carboxymethyl cellulose (CMC), 2% yeast extract, 0.2% $KH_2PO_4$, 0.03% $MnSO_4$, and 0.3% trace metal solution. Different physiological conditions, like incubation period and temperature, were also examined to assess their influence on enzyme production. Enzyme production from F. pinicola reached its highest level after cultivation for 8 days at $25^{\circ}C$. Nondenaturing polyacrylamide gel electrophoresis (PAGE), followed by the endoglucanase activity staining using CMC as the substrate, was performed to identify the endoglucanase under the culture conditions studied. Zymogram analysis of the culture supernatant revealed an endoglucanase band with a molecular mass of 52 kDa. The optimum pH and temperature for enzyme activity were $55^{\circ}C$ and pH 5.0, respectively.

Optimization of Media Composition on the Production of Melanin Bleaching Enzyme from Peniophora sp. JS17 (Peniophora sp. JS17 유래 멜라닌 탈색 효소 생산을 위한 배지 조성의 최적화)

  • Son, Min-Jeong;Kim, Yeon-Hee;Nam, Soo-Wan;Jeon, Sung-Jong
    • Microbiology and Biotechnology Letters
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    • v.47 no.2
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    • pp.250-258
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    • 2019
  • Peniphora sp. JS17, isolated from forest old tree, produced extracellular enzymes that decolorized human hair melanin. The JS17 strain had laccase and manganese peroxidase activity while it did not has lignin peroxidase activity. Batch culture indicated that the melanin decolorization activity of JS17 strain originated from laccase. The culture conditions to maximize the production of melanin bleaching enzymes from Peniophora sp. JS17 mycelia were investigated. Among the tested media for the laccase production, minimal medium (2% glucose, 0.2% malt extract, 0.1% $KH_2PO_4$, 0.4% $MgSO_4{\cdot}7H_2O$) showed the highest activity of laccase. Then, to optimize the culture condition for the laccase activity, the influence of various carbon and nitrogen sources was investigated in minimal medium. Among various carbon and nitrogen sources, 2% xylose and 0.4% tryptone showed the highest production of laccase, respectively. The enzyme was purified using $(NH_4)_2SO_4$ precipitation and Hitrap Q sepharose column, and the purified enzyme showed two isoenzymatic bands with molecular masses of about 70 kDa by SDS-PAGE. The melanin decolorization activity was 77% and 55% within 48 h in the presence of 1-hydroxybenzotriazole (HBT) and syringaldehyde, respectively, whereas only about 9% melanin decolorized in case of no mediator.

Prediction of Music Generation on Time Series Using Bi-LSTM Model (Bi-LSTM 모델을 이용한 음악 생성 시계열 예측)

  • Kwangjin, Kim;Chilwoo, Lee
    • Smart Media Journal
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    • v.11 no.10
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    • pp.65-75
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    • 2022
  • Deep learning is used as a creative tool that could overcome the limitations of existing analysis models and generate various types of results such as text, image, and music. In this paper, we propose a method necessary to preprocess audio data using the Niko's MIDI Pack sound source file as a data set and to generate music using Bi-LSTM. Based on the generated root note, the hidden layers are composed of multi-layers to create a new note suitable for the musical composition, and an attention mechanism is applied to the output gate of the decoder to apply the weight of the factors that affect the data input from the encoder. Setting variables such as loss function and optimization method are applied as parameters for improving the LSTM model. The proposed model is a multi-channel Bi-LSTM with attention that applies notes pitch generated from separating treble clef and bass clef, length of notes, rests, length of rests, and chords to improve the efficiency and prediction of MIDI deep learning process. The results of the learning generate a sound that matches the development of music scale distinct from noise, and we are aiming to contribute to generating a harmonistic stable music.

Optimization of PS-7 Production Process by Azotobacter indicus var. myxogenes L3 Using the Control of Carbon Source Composition (탄소원 조성 조절을 이용한 Azotobacter indicus var. myxogenes L3로부터 PS-7 생산 최적화)

  • Ra, Chae-Hun;Kim, Ki-Myong;Hoe, Pil-Woo;Lee, Sung-Jae;Kim, Sung-Koo
    • Microbiology and Biotechnology Letters
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    • v.36 no.1
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    • pp.61-66
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    • 2008
  • The proteins in whey are separated and used as food additives. The remains (mainly lactose) are spray-dried to produce sweet whey powder, which is widely used as an additive for animal feed. Sweet whey powder is also used as a carbon source for the production of valuable products such as polysaccharides. Glucose, fructose, galactose, and sucrose as asupplemental carbon source were evaluated for the production of PS-7 from Azotobacter indicus var. myxogenes L3 grown on whey based MSM media. Productions of PS-7 with 2% (w/v) fructose and sucrose were 2.05 and 2.31g/L, respectively. The highest production of PS-7 was 2.82g/L when 2% (w/v) glucose was used as the carbon source. Galactose showed low production of PS-7 among the carbon sources tested. The effects of various carbon sources addition to whey based MSM medium showed that glucose could be the best candidate for the enhancement of PS-7 production using whey based MSM medium. To evaluate the effect of glucose addition to whey based media on PS-7 production, fermentations with whey and glucose mixture (whey 1, 2, 3%; whey 1% + glucose 1%, whey 1% + glucose 2% and glucose 2%, w/v) were carried out. Significant enhancement of PS-7 production with addition of 1% (w/v) and 2% (w/v) glucose in 1% (w/v) whey media was observed. The PS-7 concentration of 2% glucose added whey lactose based medium was higher than that of 1% glucose addition, however, the product yield $Y_{p/s}$ was higher in 1% glucose added whey lactose based MSM medium. Therefore, the optimal condition for the PS-7 production from the Azotobacter indicus var.myxogenes L3, was 1% glucose addition to 1% whey lactose MSM medium.

Development of a Novel Medium with Chinese Cabbage Extract and Optimized Fermentation Conditions for the Cultivation of Leuconostoc citreum GR1 (폐배추 추출물을 이용한 Leuconostoc citreum GR1 종균 배양용 최적 배지 및 배양 조건 개발)

  • Moon, Shin-Hye;Chang, Hae-Choon;Kim, In-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.7
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    • pp.1125-1132
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    • 2013
  • In the kimchi manufacturing process, the starter is cultured on a large-scale and needs to be supplied at a low price to kimchi factories. However, current high costs associated with the culture of lactic acid bacteria for the starter, have led to rising kimchi prices. To solve this problem, the development of a new medium for culturing lactic acid bacteria was studied. The base materials of a this novel medium consisted of Chinese cabbage extract, a carbon source, a nitrogen source, and inorganic salts. The optimal composition of this medium was determined to be 30% Chinese cabbage extract, 2% maltose, 0.25% yeast extract, and $2{\times}$ salt stock (2% sodium acetate trihydrate, 0.8% disodium hydrogen phosphate, 0.8% sodium citrate, 0.8% ammonium sulfate, 0.04% magnesium sulfate, 0.02% manganese sulfate). The newly developed medium was named MFL (medium for lactic acid bacteria). After culture for 24 hr at $30^{\circ}C$, the CFU/mL of Leuconostoc (Leuc.) citreum GR1 in MRS and MFL was $3.41{\times}10^9$ and $7.49{\times}10^9$, respectively. The number of cells in the MFL medium was 2.2 times higher than their number in the MRS media. In a scale-up process using this optimized medium, the fermentation conditions for Leuc. citreum GR1 were tested in a 2 L working volume using a 5 L jar fermentor at $30^{\circ}C$. At an impeller speed of 50 rpm (without pH control), the viable cell count was $8.60{\times}10^9$ CFU/mL. From studies on pH-stat control fermentation, the optimal pH and regulating agent was determined to be 6.8 and NaOH, respectively. At an impeller speed of 50 rpm with pH control, the viable cell count was $11.42{\times}10^9(1.14{\times}10^{10})$ CFU/mL after cultivation for 20 hr - a value was 3.34 times higher than that obtained using the MRS media in biomass production. This MFL media is expected to have economic advantages for the cultivation of Leuc. citreum GR1 as a starter for kimchi production.