• Title/Summary/Keyword: Optimal medium

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Some Factors Affecting the Protoplast Formation and Regeneration from the Mycelium of Ganoderma lucidum(Fr.) Karsten (영지(靈芝)버섯 균사(菌絲)의 원형질체(原形質體) 나출(裸出) 및 재생(再生))

  • Shin, Gwan Chull;Yeo, Un Hyung;Yoo, Young Bok;Park, Yong Hwan
    • Korean Journal of Agricultural Science
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    • v.13 no.2
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    • pp.185-192
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    • 1986
  • Some factors affecting the protoplast release from mycelia of Ganoderma lucidum and regeneration of the protoplast were investigated and the results obtained are summarized as follows; Novozym 234 as a lytic enzyme was the most effective for the protoplast release from mycelia of Ganoderma lucidu m and its optimal concentration was 10mg per ml of osmotic stabilizer. The highest number of protoplasts were released after 3 hours incubation in the reciprocal shaking bath at 120 oscillations a minute. Among six osmotic stabilizers tested, 0.6M sucrose showed the best result. SCM medium showed good mycelial growth and high yields of protoplasts. The protoplasts released from the mycelium of G. lucidum were regenerated at 0.20 to 0.27 percent on MCM, MMM and SCM. Of the cultures obtained from protoplasts regenerated, 13 to 29 percent were monokaryon.

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Optimization of gibberellin production by Fusarium prolifertum KGL0401 and its involvement in waito-c rice growth (Fusarium prolifertum KGL0401의 지베렐린 생산 최적조건과 waito-c 생장에 미치는 영향)

  • Rim, Soon-Ok;Lee, Jin-Hyung;Lee, In-Jung;Rhee, In-Koo;Kim, Jong-Guk
    • Journal of Life Science
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    • v.17 no.1 s.81
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    • pp.120-124
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    • 2007
  • Fusarium proliferatum KGL0401 was previously isolated from Physalis alkekengi var. francheti plant roots and exhibited higher GA productivity than wild type Gibberella fujikuroi. The :tim of this work was to find out an optimal culture condition for GA production. Various carbon(fructose, glucose, lactose, maltose, sucrose) and nitrogen($KNO_3$, urea, glycine, $NaNO_3,\;NH_4Cl$) sources were used for this study. GAs activities were analysed by gas chromatography and mass spectrometry(GC-MS). The highest yield of $GA_3$ was found in the growth medium supplemented with sucrose as carbon source and $NH_4Cl$ as nitrogen source. The optimum carbon-nitrogen concentration for $GA_3$ production was found to be 0.5 M:0.17 M. Supernatant was prepared from the culture fluid of F. proliferatum KGL0401 cultured for 7 days at 3 0'E and the 10 ul supernatant was treated with 2 leaf-rice seedling.

Enhancement of Soluble Expression of Alginate Lyase By Molecular Chaperone in E. coli. (대장균에서 분자 chaperone에 의한 alginate lyase의 가용성 발현 증대)

  • Shin, Eun-Jung;Lee, Jae-Hyung;Park, So-Lim;Kim, Hyeung-Rak;Nam, Soo-Wa
    • Journal of Life Science
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    • v.17 no.1 s.81
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    • pp.132-136
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    • 2007
  • When alginate lyase gene (aly) from Pseudoalteromonas elyakovii was expressed in E. coli, most of the gene product was produced as aggregated insoluble particles known as inclusion bodies. In order to produce a soluble and active form of alginate lyase, E. coli cells fore cotransformed with the plasmids designed to permit coexpression of aly together with molecular chaperones such as DnaK/DnaJ/GrpE or GroEL/ES chaperones. The results revealed that the coexpression of aly together with DnaK/DnaJ/GrpE chaperone had a marked effect on the production of this protein as a soluble and active form, presumably through facilitating correct folding of alginate lyase protein. The optimal concentration of L-arabinose for the induction of DnaK/DnaJ/GrpE chaperone was found to be 0.05 mg/ml. When DnaK/DnaJ/GrpE chaperone was coexpressed, about 34% in the total alginate lyase was produced in the soluble fraction. By addition of 10% cetylpyridinium chloride, a clear zone around the colony coexpressing aly and DnaK/DnaJ/GrpE chaperone was formed, indicating that the alginate in the medium was hydrolyzed by active alginate lyase enzyme.

Development of Axenic Culture and Astaxanthin Production in Microalgae (미세조류를 이용한 무균분리법 개발 및 astaxanthin 생산)

  • Son, Min Chang;Lee, Dong-Jun;Park, Sejin;Kim, Min Sung;Lee, Chul Won;An, Won Gun
    • Journal of Life Science
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    • v.25 no.7
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    • pp.733-739
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    • 2015
  • Microalgae are a renewable natural resource that requires only sunlight, carbon dioxide, phosphorus, and nitrogen for rapid growth. They produce a broad variety of basic chemical substances―such as vitamins, fatty acids and carotenoids―that have high added value potential for the pharmaceutical and food industries. The aim of this study was to develop axenic culture and to establish a cell growth assay for microalgae. A further experiment was carried out to determine the yield of astaxanthin derived from microalgae. The axenic culture was developed using a mixture of antibiotics [ampicillin (100 ${\mu}g/ml$), streptomycin (10 ${\mu}g/ml$), chloramphenicol (10 ${\mu}g/ml$), penicillin (10 ${\mu}g/ml$), neomycin (50 ${\mu}g/ml$), gentamycin (50 ${\mu}g/ml$), kanamycin (10 ${\mu}g/ml$), and nystatin (1.5 ${\mu}g/ml$)] and then used to extract a variety of useful components from the microalgae. The optimal concentration for the antibiotic mixture was 1-3 percent. A spectrophotometric cell growth assay was also established. Astaxanthin was extracted from Haematococus lacustris with a yield of $1.9{\times}10^{-3}{\mu}g/l$ per 1 ml of culture medium. In conclusion, the axenic culture method developed here allows extraction of high-quality astaxanthin and other useful components from microalgae.

EFFECTS OF EXTRACTS OF DRYNARIAE RHIZOMA ON THE CHARACTERISTICS OF RAT CALARIA AND BONE MARROW CELLS (Drynariae Rhizoma추출물이 백서 두개관세포 및 골수세포 성상에 미치는 영향)

  • Lim, Kyung-Seok;Kwon, Young-Hyuk;Park, Joon-Bong;Kim, Sung-Jin;Choung, Se-Young;Park, Kun-Koo
    • Journal of Periodontal and Implant Science
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    • v.28 no.2
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    • pp.291-310
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    • 1998
  • This study was performed to evaluate the effects of extracts of Drynariae Rhizoma on the characteristics of rat calvaria cells(RCV) and bone marrow cells(RBM) which have the important role on the bone formation in vitro. Drynariae Rhizoma has been known as the useful herbal medicament for treatment of the wound healing including regeneration of bone fracture, and also has been used to treat the periodontal lesions, tooth mobility, gingival bleeding and pus discharge via sulcus in Oriental Medicine. In control group, the cells were cultured alone with Dulbeco's Modified Eagle's Medium contained with 10% fetal bovine serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. In experimental group, extracts of Drynariae Rhizoma(0.1, 1, 5, 10, $50{\mu}g/ml$) were added into the above culture condition. And then each group was characterized by examing the cell proliferation at 1, 3, 7, 14, 21, 30th day, the amount of total protein synthesis and alkaline phosphatase activity of RCV at 2,4th day and those of RBM at 3, 6th day. And also, the calcified nodule of RCV was examed at 3, 5th day in three goup, control, experimental, culture with the PDGF group. The results were as follow ; 1. Both RCV and RBM cells in Drynariae Rhizoma-treated experimental group proliferated more rapidly than nontreated control group. The experimental group below $5{\mu}g/ml$ Drynariae Rhizoma-treated showed more prominent cell proliferation from the 7th day to the 21st day than the control group and above $10\;{\mu}g/ml$ treated group in RCV. 2. Amount of total protein synthesis was more increased in Drynariae Rhizomatreated group than in control group. In $5{\mu}g/ml$ Drynariae Rhizoma-treated group showed most prominent protein synthesis of the any other exrperimental group and control group. 3. Alkaline phosphatase activity also more increased in Drynariae Rhizomatreated group than control group. 4. Mineralized nodules in Drynariae Rhizoma-treated group were more than not in control group but also in PDGF-treated group. From the above results, Drynariae Rhizoma appeared to enhanced the proliferation, protein synthesis, alkaline phosphatase activity and cellular ability of mineralized nodule formation than PDGF. So that, we conclude that Drynariae Rhizoma enhances the activities of bone cells which have the important role on the periodontal regeneration and optimal application of Drynariae Rhizoma was thought to be useful as the means in bone regeneration.

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Effects of platelet-derived growth factor and epidermal growth factor on the characteristics of beagle dog's periodontal ligament and bone marrow cells (혈소판유래성장인자와 상피성장인자가 치주인대세포와 골수세포의 성상에 미치는 영향)

  • Cho, Byeong-Do;Herr, Yeek;Park, Joon-Bong;Kwon, Young-Hyuk;Lee, Man-Sup
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.491-510
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    • 1996
  • This study was performed to evaluate the effects of platelet-derived growth factor (PDGF) and epidermal growth factor (EGF) on the characteristics of beagle dog's periodontal ligament (BPD) cells and bone marrow (BBM) cells which have the important role on the early stage of periodontal tissue regeneration in vitro. In control group, the cells ($1.5{\times}10^5$cells/ml) were cultured alone with Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum, $50{\mu]g/ml$ ascorbic acid, and 10mM/ml ${\beta}-glycerophosphate$. In experimental groups, growth factors, PDGF or EGF(10ng/ml), were added into the above culture condition. And then each group was characterized by examining the cell proliferation rate, amount of total protein synthesis, alkaline phosphatase activity at 1, 5, 9, 13, 17th day after seeding of cells into the culture wells. The results were as follows: 1. Both BPD and BBM cells in PDGF-treated group proliferated more rapidly than non-treated cells. This finding also was observed in EGF-treated group but it was not as prominent as that of PDGF-treated group. The proliferation rates of both cells showed the time-dependent pattern during experimental periods in all three groups. 2. Amount of total protein synthesis was more increased in PDGF-treated group than in control group. But no significant difference between EGF-treated group and control group was observed throughout experimental periods even though the tendency of amount of protein synthesis was time-dependent pattern. 3. Alkaline phosphatase activity also more increased in PDGF-treated group than control group. But slight decrease tendency was seen in both cells of EGF-treated group. From the above results, PDGF appeared to enhance the proliferation and cellular activities including amount of total protein synthesis and alkaline phosphatase activity of BPD and BBM cell, but EGF did not show notable effects. The optimal application of these growth factors was thought to be useful as the adjunctive means in periodontal regeneration procedures.

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Suitability of Various Domestic Wheats for Korean-Style Steamed Bread (한국형 찐빵 제조시 국내산 밀 품종의 적합성)

  • 김창순;황철명;김혁일;정동진;한재홍
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.6
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    • pp.1129-1136
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    • 2001
  • This study was to investigate the suitability of six domestic wheat cultivars for the Korean-style steamed bread made under optimal conditions. Six wheat flours milled from cultivars of Greu, Kumgang, Eunpa, Taptong Kobun, and Allgreu contained 13.8, 13.7, 13.7, 13.0, 11.7, 11.0% of protein. Control bread was made from blend (protein 10.5%) of 50% high strength and 50% low strength wheat flours milled from imported wheats. The volume of steamed bread made from Kumgang was highest followed by Eunpa, Tapdong, Kobun, Greu, control bread, Allgreu. Especially, the bread qualities of Kumgand and Kobun were superior to the control bread, showing better surface characteristics such as smoothness, glossiness, and whiteness, better shapes and desirable texture. Domestic wheat flours, having medium strength with high protein content above 13.0% were suitable for steamed bread except for Greu. Volumes of steamed bread made from domestic wheat flours were correlated with protein and ash content, flour color (L value), farinograph dough development time and stability, whereas spread ratio, total bread score and overall acceptability were correlated with farinograph dough stability It is concluded that flour quality is more important factor than protein content when domestic wheat flours are chosen for Korean-style steamed bread

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Molecular Weight Distribution Inside and Outside Capsules Using Coencapsulating Technology (공동캡슐화를 이용한 Capsule 내외부의 분자량 분포)

  • 이기선;임현수
    • KSBB Journal
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    • v.16 no.4
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    • pp.321-326
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    • 2001
  • The change of molecular weight inside and outside a capsule produced using coencapsulating technology was investigated. Chitosan and chitosanase were enveloped in this membrane and product released was a loaded the medium by the principle of size exclusion. The leakage of substrate corresponding to the agitation speed was controlled by adjusting the alginate and CaCO$_3$ concentrations. The optimal condition of alginate concentration and agitation speed were 0.5% and 40rpm, respectively. Membrane thickness and capsules diameter were 10 $\mu$m and approx. 3.0 - 1.5 mm, respectively. Molecular weight difference by concentration and alginate viscosity were of little significance. In accordance with the molecular weight distribution versus enzyme concentration relationship, low concentration of enzyme produced high molecular weight oligosaccharides. At a 1.5 mm capsule size the product diffusion rate to outer surface highest. The molecular weight distribution of the released oligosaccharides was ranged from 1000 to 6000 Da. More than 80% of the initial activity of encapsulated enzyme retained after 8hrs of reaction.

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A Study on the Optimum Conditions of Gelatin-Degrading Proteolytic Enzyme Production from Bacillus subtilis B0021 (Bacillus subtilis B0021가 생산하는 Gelatin 분해성 Proteolytic Enzyme 생산의 최적의 연구)

  • 백대헌;이항우복성해
    • KSBB Journal
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    • v.10 no.4
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    • pp.374-385
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    • 1995
  • Nutritional requirements and cultural conditions for the production of extracellular gelatin-degrading proteolytic enzyme by Bacillus subtilis B0021 were investigated. Optimum carbon source for proteolytic enzyme production was salicin, but it was substituted by glucose for economical reason. The fermentation medium giving a maximum proteolytic enzyme activity was consisted of 1.5%(w/v) glucose, 2.5%(w/v) yeast extract, and 0.001%(w/v) manganese sulfate and 0.002%(w/v) ferrous sulfate. Proteolytic enzyme activity of B. subtilis B0021 was completely inhibited by 0.5%(w/v) tannic acid. Initial pH was optimal at 7.0 and the enzyme activity in the flask culture usually reached a maximal level after 36 hours of fermentation at $30^{\circ}C$. In the $5\ell$ fermentor fermentation at $30^{\circ}C$, enzyme activity was maximum at 36 hour of cultivation but after this enzyme activity was decreased rapidly. Initial viscosity of 45%(w/v) gelatin(2,800mPas) was decreased rapidly to 96%(mPas) after hydrolysis for 4hr at $40^{\circ}C$ by crude enzyme of B. subtilis B0021.

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A Study on the Mitigation of the Exposure Dose Applying Bolus Tracking in Brain Perfusion CT Scan (뇌 관류 CT검사에서 BolusTracking기법을 적용한 피폭선량 저감화에 관한 연구)

  • Kim, Ki-Jeong;Jung, Hong-Ryang;Lim, Cheong-Hwan;Hong, Dong-Hee;Shim, Jae-Goo;You, In-Gyu
    • Journal of Digital Convergence
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    • v.12 no.3
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    • pp.353-358
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    • 2014
  • This study was conducted to analyze the patient's exposed dose targeting the patients who had acute ischemic stroke symptoms and CT brain perfusion scan, by comparing fixed time technique and bolus tracking technique which was provided by the manufacturer and to identify the Time graph to implement the usability of contrast medium's tracking technique the best contrast enhancement intervals. $CTDI_{VOL}$ of PCT in patient appeared to be 431.72mGy in fixed scan delay protocol, whereas 323.61mGy in Bolus tracking technique. The value of DLP appeared to be $1243.47mGy{\cdot}cm$ in fixed scan delay protocol, whereas $932mGy{\cdot}cm$ in Bolus tracking technique. Time graph appeared to be various in fixed scan delay protocol, whereas the optimal time graph could be obtained in Bolus tracking. The exposure dose could be reduced by 25% applying Bolus tracking technique when taking brain perfusion CT scan.