• Title/Summary/Keyword: Mass culture conditions

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Production of Medium-chain-length Poly (3-hydroxyalkanoates) by Pseudomonas sp. EML8 from Waste Frying Oil (Pseudomonas sp. EML8 균주를 이용한 폐식용류로부터 medium-chain-length poly(3-hydroxyalkanoates) 생합성)

  • Kim, Tae-Gyeong;Kim, Jong-Sik;Chung, Chung-Wook
    • Journal of Life Science
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    • v.31 no.1
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    • pp.90-99
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    • 2021
  • In this study, to reduce the production cost of poly(3-hydroxyalkanoates) (PHA), optimal cell growth and PHA biosynthesis conditions of the isolated strain Pseudomonas sp. EML8 were established using waste frying oil (WFO) as the cheap carbon source. Gas chromatography (GC) and GC mass spectrometry analysis of the medium-chain-length PHA (mcl-PHAWFO) obtained by Pseudomonas sp. EML8 of WFO indicated that it was composed of 7.28 mol% 3-hydrxoyhexanoate, 39.04 mol% 3-hydroxyoctanoate, 37.11 mol% 3-hydroxydecanoate, and 16.58 mol% 3-hydroxvdodecanoate monomers. When Pseudomonas sp. EML8 were culture in flask, the maximum dry cell weight (DCW) and the mcl-PHAWFO yield (g/l) were showed under WFO (20 g/l), (NH4)2SO4 (0.5 g/l), pH 7, and 25℃ culture conditions. Based on this, the highest DCW, mcl-PHAWFO content, and mcl-PHAWFO yield from 3-l-jar fermentation was obtained after 48 hr. Similar results were obtained using 20 g/l of fresh frying oil (FFO) as a control carbon source. In this case, the DCW, the mcl-PHAFFO content, and the mcl-PHAFFO yields were 2.7 g/l, 62 wt%, and 1.6 g/l, respectively. Gel permeation chromatography analysis confirmed the average molecular weight of the mcl-PHAWFO and mcl-PHAFFO to be between 165-175 kDa. Thermogravimetric analysis showed decomposition temperature values of 260℃ and 274.7℃ for mcl-PHAWFO and mcl-PHAFFO, respectively. In conclusion, Pseudomonas sp. EML8 and WFO could be suggested as a new candidate and substrate for the industrial production of PHA.

Cultivation of Lactic Acid Bacteria for the Development of Probiotic Products using Red Ginseng Starch (프로바이오틱스 개발을 위한 홍삼 전분을 활용한 유산균 배양)

  • Kim, Yeong-Su;Lee, Hwan;Kim, Do-Yeon;Kim, So-Young;Lee, Wan-Kyu;Lee, Sang-Myeong;Park, Jong-Dae;Shon, Mi-Yae
    • Journal of the East Asian Society of Dietary Life
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    • v.23 no.6
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    • pp.818-826
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    • 2013
  • To reduce the production cost of Lactobacillus, discarded red ginseng starch was collected from a factory of red ginseng extract in order to develop the Lactobacillus culture medium. According to the analysis of the gensenoside composition of red ginseng starch, the total gensenoside content of starch was 2.73 mg/g, and the gensenoside $Rb_1$, $Rb_2$ and $Rg_3$ contents were 0.1, 0.29 and 0.52 mg/g, respectively. For the preparation of the liquid media, red ginseng starch was added at rates of 0, 5, 10 and 20%. Further, Lactobacillus plantarum 15357 and Leuconostoc mesenteroides sub sp. strains were then inoculated to these prepared broths. With the red ginseng starch medium, the growth rates ($1.42{\times}10^7$ and $2.96{\times}10^{10}$ CFU/mL) and the final cell concentrations were higher than the MM medium ($1.0{\times}10^7$ CFU/mL). The optimal concentration of red ginseng starch and yeast extract as a medium were 20% and 10 g/L, respectively. Under these conditions, the cell mass of L. plantarum 15357 and L. mesenteroides sub sp. reached $5.11{\times}10^{10}$ and $8.17{\times}10^{10}$ CFU/mL. These results show a great possibility for the utilization of red ginseng starch as economic medium sources in the production of cell mass of lactic acid bacteria. This is the first trial of development of economic LAB growth medium using discarded red ginseng starch.

Mass-Production of Acetylcholinesterase Sensitive to Organophosphosphates and Carbamates Insecticides (유기인계 및 카바메이트계 농약의 고감수성 아세틸콜린에스테라이즈의 대량생산)

  • Kim, Young-Mee;K., Cho;Cho, Moon-Jae
    • Applied Biological Chemistry
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    • v.46 no.4
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    • pp.353-360
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    • 2003
  • For the simple rapid bioassay of organophosphorus and carbamate pesticide residues, a mass-production system of acetycholinesterase (AChE, EC 3.1.1.7, MAChE) using baculovirus and insect cell culture was constructed. The cDNA for AChE was synthesized from Drosophila melanogaster in Halla Mountain, the lipid anchor tail was removed by PCR and was used for the site-directed mutagenesis of three amino acid residues (E107Y, F368L, L408F). The mutated cDNA was inserted into the baculovirus vector and expressed in insect cells. Maximum cell growth and enzyme activity were reached when the cells $(2{\times}10^6\;cell/ml)$ were infected four times at four-day-intervals. His-tag containing MAChE was purified using Ni-NTA column and used for characterization. The activity was maintained under various pHs (3-10) and temperatures $(20-50^{\circ}C)$ under experimental conditions. As an extraction solution for pesticides, methanol is more effective than ethanol. Against major organophosphate and carbamate pesticides, the MAChE showed better sensitivity than AChE and AChE from housefly (Taiwan).

A News Frame Analysis by the South Korean Press on the Livelihoods of a North Koreans (북한주민 생활 실태에 관한 국내 신문보도 프레임연구: 조선일보, 동아일보, 한겨레, 경향신문을 중심으로)

  • Ha, Seung-Hee;Lee, Min-Kyu
    • Korean journal of communication and information
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    • v.58
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    • pp.222-241
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    • 2012
  • Analysis research was conducted on the 'Chosen Ilbo,' 'Donga Ilbo,' 'Hankyoreh' and 'Kyunghyang Newspaper' on news reporting frame and sources regarding the North Korean population's living and status of their respective conditions throughout South Korean administrations of Kim Dae-jung, Roh Mu-hyuen, and Lee Myung-Bak. Resulting first and second frame analyses showed that the four newspapers showed clear distinctions in their execution and attitude in their reports in accordance to the philosophies of each 'progressive/left-wing newspapers' and 'conservative/right-wing newspapers'. Moreover, the first and second analyses showed that the reports mimicked each individual presidential administration and their political policy regardless of the newspaper's political voice. In terms of the usage of 'anonymous sources,' conservative newspapers were found to use them more frequently than their counterpart, the liberal newspapers. In conclusion to the overall analyses, with the continued tension between the International community and North Korea, the reports of North Korean inhabitant activities are inevitably portrayed in a negative tone in effort to plant a distorted view to the South Korean citizens. Furthermore, this manipulation of the press may affect the credibility of South Korean press in terms of the topic of North Korean culture.

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Bacillus subtilis를 이용한 대두 발효식품의 혈전용해능

  • Jeong, Yeong-Gi
    • Proceedings of the Korean Society of Life Science Conference
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    • 2001.06a
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    • pp.67-86
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    • 2001
  • A strain producing strongly fibrinolytic enzyme was isolated from soil and was identified to be Bacillus subtilis by biochemical and physiological characterization. The optimal culture conditions for the production of fibrinolytic enzyme was determined to be 1.0% tryptone, 1.5% soluble starch, 0.5% Peptone, 0.5% NaCl, $(NH_{4})_{3}PO_4.3H_{2}O, and MgSO_{4}.7H_{2}O.$ Initial pH and temperature were pH 8.0 and $30^{\circ}C$ , respectively, The highest enzyme production was observed at 30 hours of cultivation at $30^{\circ}C$ The fibrinolytic enzyme was purified to homogeneity by DEAE Sephadex A-50 ion exchange column chromatography, 70% ammonium sulfate precipitation, Sephadex G-200 and G-75 gel filtration column chromatography. The molecular weight of the purified enzyme was 28,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A gene encoding the fibrinolytic enzyme was cloned into a plasmid vector pBluescript, transforming E.coli XL-1 Blue. The clone was able to degrade fibrin, This indicated that the gene could encode a fibrinolytic enzyme. The nucleotide sequence of the 2.7 kb insert was determined in both direction. One open reading frame composed of 1023 nucleotides was found to be a potential protein coding region. There was the putative Shine-Dalgano sequence and TATA box upstream of the open reading frame. The homology search data in the genome database showed that both the 2.7 kb insert and 1 kb open reading frame carried no significance in the nucleotide sequence of known fibrinolytic enzyme from Bacillus serovars. The recombinant cell harboring the novel gene involved in fibrinolysis was subjected to protein purification. The molecular mass of the purified fibrinolytic enzyme was determined to be 31864 Dalton, which was highly in accordance with the molecular mass(33 kDa) of the fibrinolytic gene deduced from the insert. The fibrinolytic enzyme was Purified 50.5 folds to homogeneity in overall yield of 10.7% by DEAE Sephadex A-50 ion exchange, 85% ammonium sulfate precipitation, Sephadex G-50, Superdex 75 HR FPLC gel filtration. In conclusion, a novel fibrinolytic gene from Bacillus subtilis was identified and characterized by cloning a genomic library of Bacillus subtilis into pBleuscript. For the soybean fermented by this strain, it is found that there increased assistant protein about 20% compared to the soybean not fermented and increased about 30% according to amino acid analysis and, in particular, essential amino acid increased about 40%. When keeping this fermented soybean powder at room temperature for about 70days, it showed very high stability maintaining almost perfect activity and, therefore, it gave us great suggestion its possibility of development as a new functional food.

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Mass Cultivation and Characterization of Multifunctional Bacillus velezensis GH1-13 (복합기능성 Bacillus velezensis GH1-13 균주의 대량배양 최적화 및 특성)

  • Park, Jun-Kyung;Kim, JuEun;Lee, Chul-Won;Song, JaeKyeong;Seo, Sun-Il;Bong, Ki-Moon;Kim, Dae-Hyuk;Kim, Pyoung Il
    • Korean Journal of Organic Agriculture
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    • v.27 no.1
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    • pp.65-76
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    • 2019
  • Bacillus genus are found abundantly in various sites and their secondary metabolites were used as potential agents in agriculture, notably plant growth promoting and bio-control. The objective of this study was to develop the culture conditions of GH1-13 strain including higher cell growth, stable endospore-forming and enhancement of potential agents which are related with plant growth promoting and phytopathogen suppression. The optimal carbon and nitrogen sources were determined by glucose and soy bean flour, respectively, then resulted in $7.5{\times}10^9cells/mL$, $6.8{\times}10^9\;endospore\;cells/mL$ and sporulation yield of 90% after 30 h cultivation in 500 L submerged fermenter at $37^{\circ}C$, pH 7.0. Cells and cell-free supernatant of GH1-13 strains showed the potent antifungal activity against phytopathogenic fungi of Colletotrichum gloeosporioides. It was also confirmed that indole-3-acetic acid (IAA) production of GH1-13 strain was greatly increased by addition of 0.3% tryptophan.

Seawater Temperature Variation at Aquafarms off Wando in the Southwest Coast of Korea (완도 양식장 해역의 수온변동)

  • Yang, Joon-Yong;Lee, Joon-Soo;Han, In-Sung;Choi, Yong-Kyu;Suh, Young-Sang
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.18 no.6
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    • pp.514-519
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    • 2012
  • Abalone culture is one of important coastal fisheries off Wando in the southeast coast of Korea. Since cage culture for abalones was popularized, Understanding of temperature variation, which is important to raise abalones and to prevent their mass mortalities, is necessary. We analyzed temperature data from 2005 to 2009 obtained at Sinji-do and Cheongsan-do off Wando. Sinji-do, which is relatively close to land, had yearly wide range of temperature and rate of temperature variation. It is likely to be caused by heating of solar radiation in summer and cooling in winter at the shallow area. Rate of temperature variation in autumn was higher than that in spring. In summer short term variation of temperature corresponding tidal period was distinguished clearly. Diurnal temperature range, abrupt temperature change, was larger open sea. Comparison between temperatures of two stations and favorable raising conditions of abalones showed that Cheongsan-do, located out to sea, appears to be more appropriate than Sinji-do.

Wastewater Utilization: A Place for Managed Wetlands - Review -

  • Humenik, F.J.;Szogi, A.A.;Hunt, P.G.;Broome, S.;Rice, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.12 no.4
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    • pp.629-632
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    • 1999
  • Constructed wetlands are being used for the removal of nutrients from livestock wastewater. However, natural vegetation typically used in constructed wetlands does not have marketable value. As an alternative, agronomic plants grown under flooded or saturated soil conditions that promote denitrification can be used. Studies on constructed wetlands for swine wastewater were conducted in wetland cells that contained either natural wetland plants or a combination of soybeans and rice for two years with the objective of maximum nitrogen reduction to minimize the amount of land required for terminal treatment. Three systems, of two 3.6 by 33.5 m wetland cells connected in series were used; two systems each contained a different combination of emergent wetland vegetation: rush/bulrush (system 1) and bur-reed/cattail (system 2). The third system contained soybean (Glycine max) in saturated-soil-culture (SSC) in the first cell, and flooded rice (Oryza sativa) in the second cell. Nitrogen (N) loading rates of 3 and $10kg\;ha^{-1}\;day^{-1}$ were used in the first and second years, respectively. These loading rates were obtained by mixing swine lagoon liquid with fresh water before it was applied to the wetland. The nutrient removal efficiency was similar in the rush/bulrush, bur-reed/cattails and agronomic plant systems. Mean mass removal of N was 94 % at the loading rate of $3kg\;N\;ha^{-1}\;day^{-1}$ and decreased to 71% at the higher rate of $10kg\;N\;ha^{-1}\;day^{-1}$. The two years means for above-ground dry matter production for rush/bulrushes and bur-reed/cattails was l2 and $33Mg\;ha^{-1}$, respectively. Flooded rice yield was $4.5Mg\;ha^{-1}$ and soybean grown in saturation culture yielded $2.8Mg\;ha^{-1}$. Additionally, the performance of seven soybean cultivars using SSC in constructed wetlands with swine wastewater as the water source was evaluated for two years, The cultivar Young had the highest yield with 4.0 and $2.8Mg\;ha^{-1}$ in each year, This indicated that production of acceptable soybean yields in constructed wetlands seems feasible with SSC using swine lagoon liquid. Two microcosms studies were established to further investigate the management of constructed wetlands. In the first microcosm experiment, the effects of swine lagoon liquid on the growth of wetland plants at half (about 175 mg/l ammonia) and full strength (about 350 mg/l ammonia) was investigated. It was concluded that wetland plants can grow well in at least half strength lagoon liquid. In the second microcosm experiment, sequencing nitrification-wetland treatments was studied. When nitrified lagoon liquid was added in batch applications ($48kg\;N\;ha^{-1}\;day^{-1}$) to wetland microcosms the nitrogen removal rate was four to five times higher than when non-nitrified lagoon liquid was added. Wetland microcosms with plants were more effective than those with bare soil. These results suggest that vegetated wetlands with nitrification pretreatment are viable treatment systems for removal of large quantities of nitrogen from swine lagoon liquid.

Effect of Growth Regulator, Sucrose, and Minimal-growth Conservation on In Vitro Propagation of Virus-free Sweet Potato Plantlets (고구마 무병묘의 기내 증식에 미치는 생장조절물질, Sucrose, 최소생장 보존의 영향)

  • Lee, Na Rha;Lee, Seung Yeob
    • Journal of Bio-Environment Control
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    • v.29 no.1
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    • pp.1-8
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    • 2020
  • The influence of growth regulators (NAA and BA) and sucrose concentrations (0, 3, 5, 7, 9%) on in vitro rapid-propagation of virus-free sweet potato [Ipomoea batatas (L.) Lam.] was investigated with single-node or shoot-tip culture of two cultivars ('Matnami' and 'Shinhwangmi'). The survival rate and growth of shoot-tip explant was also investigated under the presence or absence of light (blue and red LED = 7:3, 150±5 μmol·m-2·s-1 PPFD) during minimal-growth in vitro conservation at 15℃. Vine length, vine diameter, fresh weight and dry weight were enhanced without callusing of explant in the MS medium supplemented with 0.2-0.5 mg·L-1 BA. The growth of single-node and shoot-tip explants were significantly enhanced with the increase of vine length, number of leaf, number of root, fresh weight, and dry weight in the solid medium containing 5% sucrose and 0.2 mg·L-1 BA. Vine elongation of shoot-tip explants were highest in the liquid medium containing 3% sucrose than the solid medium. The survival rate of minimal-growth in vitro conservation was 100% in 5 months under the presence of light (LED, 150±5 μmol·m-2·s-1 PPFD) at 15℃, but the explants in dark condition died in 3 months. The light was absolutely necessary for the in vitro conservation under minimal-growth conditions of virus-free sweet potato plantlets at 15℃, and the high density of explants (10 plantlets per Petri Dish) was increased the efficiency of mass conservation.

Establishment of Callus Induction and Plant Regeneration System from Mature Seeds of Miscanthus sinensis (억새(Miscanthus sinensis) 성숙 종자로부터의 캘러스 유도 및 식물체 재분화 체계 확립)

  • Cho, Joon-Hyeong;Byeon, Ji-Hui
    • Korean Journal of Plant Resources
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    • v.24 no.5
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    • pp.628-635
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    • 2011
  • This study was conducted to establish the tissue culture system for Korean domestic Miscanthus sinensis, which is used in various purposes such as forage, and bio-energy resources. With the mature seed of Miscanthus, optimum concentrations of plant growth regulators were identified for an efficient callus induction and regeneration. Among the treatments of 1~10 $mg{\cdot}L^{-1}$ 2,4-D, IBA, or NAA, callus induction rate was highest (85.3%) on MS medium containing 5 $mg{\cdot}L^{-1}$ 2,4-D. Under the condition, the callus were efficiently induced and proliferated with comparably lower frequencies of callus browning. In shoot regeneration, the treatment of NAA combined with BAP seemed to contribute more efficient conditions to shoot regeneration than those of NAA with Kinetin or 2-iP. Especially, regeneration efficiency and number of regenerated plants were 83.7% and 5.5 in 3 $mg{\cdot}L^{-1}$ NAA with 5 $mg{\cdot}L^{-1}$ BAP, respectively, which were higher frequencies than those in NAA with Kinetin or 2-iP. In results, 5 $mg{\cdot}L^{-1}$ 2,4-D and 3 $mg{\cdot}L^{-1}$ NAA combined with 5 $mg{\cdot}L^{-1}$ BAP were efficient for embryogenic callus induction and regeneration of Miscanthus. This system would be useful for mass-propagation and developing new cultivars via tissue culture of Miscanthus sinensis.