• Title/Summary/Keyword: 분리조

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Eotaxin mRNA Expression in Bronchial Mucosa of Patients with Asthma (천식 환자의 기관지 조직에서 Eotaxin mRNA 발현에 관한 연구)

  • In, Kwang-Ho;Cho, Jae-Yun;Kang, Sae-Yong;Lee, Sang-Youb;Shim, Jae-Jeong;Kang, Kyung-Ho;Yoo, Se-Hwa;Na, Young-Soon;Kim, Han-Gyum
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.697-704
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    • 1998
  • Background: Asthma is a chronic inflammatory disease of the airways characterized by a marked infiltration of eosinophils in the bronchial mucosa. Asthmatic bronchial mucosa produces many factors described as being chemotactic for inflammatory cells. IL-5, RANTES, and MCP-1 alpha are the chemotactic factors for eosinophils, but their roles are controversial. Recently eotaxin that is a potent eosinophil chemoattractant cytokine was detected in a guinea-pig model of allergic airway inflammation, and human eotaxin was cloned. Eotaxin is a specific chemoattractant for eosinophils, but its role in asthma is not confirmed. We examined the in vivo expression of eotaxin in bronchi of asthmatic patients. Methods : 11 asthmatics and 2 normal controls were enrolled. All subjects were underwent bronchoscopy with bronchial biopsies in 2nd or 3rd carina. RNA extraction from biopsy samples was done by acid-guanidium method. Semi-quantitaive RT-PCR was done for evaluation of eotaxin mRNA expression The extent of eosinophil infiltration was evaluated by counting the eosinophils in submucosa in HPF of microscope. Results : Eotaxin mRNA expressed in symptomatic, uncontrolled asthma. Steroid inhibited expression of eotaxin mRNA in asthma. Expression of eotaxin mRNA correlated with eosinophil infiltration in bronchial tissues. Conclusion: Expression of eotaxin mRNA increases in uncontrolled asthma and eotaxin is involved in the recruitment of eosinophils.

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The Immunohistochemical Analysis for the Expression of Survivin, an Inhibitor of Apoptosis Protein, in Non-small Cell Lung Cancer (비소세포폐암에서 아포프토시스 억제 단백질 Survivin 발현에 관한 면역조직학적 분석)

  • Ko, Mi-Hye;Myoung, Na-Hye;Lee, Jae-Whan;Cho, Eun-Mi;Park, Jae-Seuk;Kim, Keun-Youl;Lee, Kye-Young
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.6
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    • pp.909-921
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    • 2000
  • Background : Defects in apoptotic signaling pathways play important role in tumor initiation, progression, metastasis and resistance to treatment. Several proteins which may promote tumorigenesis by inhibiting apoptosis were identified. The survivin protein is the member of inhibitor of apoptosis protein(IAPs) family which inhibits apoptosis. Unlike other IAPs, it is expressed in during the fetal period but not in adult differentiated tissues. Many reports have stated that survivin is selectively expressed in many cancer cell lines and cancer tissues. We performed immunohistochemical analysis for survivin expression in non-mall cell lung cancer to get evaluate its clinical implication. Methods : Twenty nine surgically resected lung cancers were examined. Immunohistochemical staining were performed by immuno-peroxidase technique using avidin-biotinylated horseradish pemxidase complex in the formalin-fixed, paraffin-embedded tissue $4{\mu}m$ section. Anti-survivin polyclonal antibody was used for primary antibody and anti-p53 monoclonal antibody was also used to analyze the correlation between survivin and p53 expression. The survivin expression scores were determined by as the sum of the stained area and intensity. Results : Immunohistochemical analysis showed cancer specific expression of survivin in 20 of 29 cases (69.0%). Western blot analysis also showed the selective survivin expression in tumor tissue. There was no correlation between survivin expression and clinicopathological parameters and prognosis. We analyzed the ∞π'elation between survivin expression and p53 expression, but found none. Conclusion: We confirmed the tumor specific expression of survival in non-small cell lung canær. But this expression was not correlated with clinical parameters as well as histology, tumor stage, recurrence, and survival rate. Also it was not statistically correlated with the expression of p53.

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Loss of the Retinoblastoma Gene in Non-Small Cell Lung Cancer (비소세포폐암에서의 망막모세포종유전자의 소실)

  • Lee, Choon-Taek;Kim, Chang-Min;Zo, Jae-Ill;Shim, Young-Mog;Hong, Weon-Seon;Lee, Jhin-Oh;Kang, Tae-Woong
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.2
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    • pp.98-103
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    • 1993
  • Background: Inactivation of retinoblastoma gene (Rb) has been observed in a variety of human cancers. Loss of heterozygosity (LOH) of Rb which is a common mode of allelic inactivation of Rb, has been known as a frequent genetic event in small cell lung cancer but it has been detected less frequently in non-small cell lung cancer. To define the role of Rb deletion in lung cancer, we investigated the genomic DNAs of 43 non-small cell lung cancers and 1 small cell lung cancer paired with normal lung tissues obtained by thoracotomy. Methods: The genomic DNAs were obtained by the digestion with proteinase K followed by phenol-chloroform extraction method. The genomic DNAs were digested by restriction endonuclease (EcoRI), separated by agarose gel electrophoresis, transferred to nylon membrane by Southern blot transfer and then hybridized with labelled Rb 1 probe which contains. 1.4 kb sized DNA sequence containing N-terminal portion of Rb. Results: In 26 squamous cell lung cancers, 16 cases were informative after EcoRI digestion and LOH of Rb was found in 10 cases (62.5%). In 17 adenocarcinomas of lung, 11 cases were informative and LOH of Rb was found in five cases (45.4%). The analysis of clinical parameters revealed no significant differences between the two groups with or without LOH of Rb in the aspects of age, sex, degree of differentiation, stage and smoking amount. Conclusions: These results suggest that Rb inactivation is also significantly involved in the molecular pathogenesis of non-small cell lung cancer.

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Matrix metalloproteinase-1 suppression and type-1 procollagen synthesis promoting effects of Uncaria gambir (아선약 추출물의 matrix metalloproteinase-1(MMP-1) 억제 및 제1형 프로콜라겐 합성 촉진 활성)

  • Lee, Jaemin;Kim, Dong-Hee;Kim, Eun-Woo;Kwon, Hyun Ju;Kim, Byung Woo;Cho, Jae-Hyon;Kim, Hyun-Joo;Kim, Tae Hoon
    • Food Science and Preservation
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    • v.24 no.1
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    • pp.93-99
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    • 2017
  • Collagen synthesis is decreased and matrix metalloproteinase-1 (MMP-1) levels are increased in naturally aged human skin, and these alterations cause changes such as skin wrinkling and decreased elasticity. As a part of our ongoing search for bioactive ingredients, MMP-1 inhibitory and type-1 procollagen synthesis inducing activities of aqueous methanolic extract of manufactured gambir product from Uncaria gambir were investigated in in vitro bioassay systems. In addition, total phenolic contents were quantified using a spectrophotometric method. Among tested samples, 40% MeOH eluate from 80% methanolic extract of manufactured U. gambir using open column chromatography packed with Diaion HP-20 resin showed significant MMP-1 inhibitory activities with an $IC_{50}$ value of $15.6{\pm}1.3{\mu}g/mL$. Furthermore, type-1 procollagen synthesis promoting property of 40% MeOH eluate ($IC_{50}$ value; $6.9{\pm}0.7{\mu}g/mL$) from 80% methanolic extract of manufactured gambir was higher than other eluates. Additionally, the present investigation revealed that 40% MeOH eluate of manufactured gambir product contained a high level of total phenolic compounds. The result suggests a distinct relationship between anti-wrinkle activity and total phenolic contents, and manufactured gambir product could be considered a new effective source of natural bioactive ingredients. Systematic investigation of manufactured gambir product will be performed for further development of its biological properties.

Studies on the Enzyme from Arthrobacter luteus Accelerating the Lysis of Yeast Cell Walls - I. Effects of Various Factors on the Lysis of Yeast Cell Walls by the Preparation of Crude Zymolyase (Arthrobacter luteus가 생산(生産)하는 효모(酵母) 세포벽(細胞壁) 용해(溶解) 촉진(促進) 효소(酵素)에 관(關)한 연구(硏究) - 제(第) 1 보(報) : Zymolyase 조(粗) 효소(酵素)에 의한 효모(酵母) 세포벽(細胞壁) 용해(溶解)에 미치는 제(諸) 인자(因子)의 영향(影響) -)

  • Oh, Hong-Rock;Shimoda, Tadahisa;Funatsu, Masaru
    • Korean Journal of Food Science and Technology
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    • v.11 no.4
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    • pp.242-248
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    • 1979
  • To detect proper lytic assay conditions of the crude zumolyase from Arthrobacter luteus, effets of the various factors involved in the lytic system of Sacchromyces sake cultured with shaking in the malt extracts medium were investigated. The results are summarized as follows : 1. The susceptibilities of viable cells of S. sake from logarithmic growth phase to the lytic enzmye were much greater than those of the cells in lag and stationary phases. The cells cultured for 18 hr were the most susceptible to the enzyme. 2. Lytic activity of the enzyme toward the viable cells of S. sake was very low. It was, however, enhanced 4 folds of more by the pretreatment of the cells with 0.05 M sodium sulfite. 3. Lytic activity of the enzyme toward commercial baker's yeast cells was negligible, and the effect of sodium sulfite on the lysis of the cells also was nothing but a little. 4. The lyophilized cells of the baker's yeast showed more susceptibility to the lytic enzyme than viable cells of the yeast. No definite effect of sodium sulfite on the lysis of the lyophilized cells, however, was observed either baker's yeast of S. sake. 5. It appeared that the relationship between the reaction rate and the enzyme concentration on the lysis of the yeast cell walls followed enzyme kinetic theory, but one between the reaction rate and concentration of the yeast cells as substrates showed different pattern from that in enzyme kinetic theory. 6. After the preparation of crude zymolyase was kept at $7^[\circ}C$ for 10 days in the 0.05 M phosphate buffer, pH 7.5, the remainning lytic activity was about 80 %.

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Expression and Purification of Recombinant Human Interferon-gamma Produced by Escherichia coli (대장균이 생산한 재조합 인체 감마인터페론의 발현과 정제)

  • Park, Jung-Ryeol;Kim, Sung-Woo;Kim, Jae-Bum;Jung, Woo-Hyuk;Han, Myung-Wan;Jo, Young-Bae;Jung, Joon-Ki
    • KSBB Journal
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    • v.21 no.3
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    • pp.204-211
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    • 2006
  • For the production of the recombinant human interferon-gamma(rhIFN-${\gamma}$) in Escherichia coli, human glucagon and ferritin heavy chain were used as fusion partners. Even though rhIFN-${\gamma}$ is expressed as an inclusion body form in E. coli because of strong hydrophobicity of itself, over 50% of fused rhIFN-${\gamma}$ was expressed as soluble form in E. coli $Origami^{TM}$(DE3) harboring pT7FH(HE)-IFN-${\gamma}$ which encodes ferritin heavy chain-fused rhIFN-${\gamma}$. In the case of using glucagon-ferritin heavy chain hybrid mutant as a fusion partner, 6X His-tag was additionally introduced to N-terminus of GFHM(HE)-IFN-${\gamma}$ for enhancing purification yields of rhIFN-${\gamma}$. Fusion protein HGFHM(HE)-IFN-${\gamma}$ with two 6X His-tag was more effectively bound to Ni-NTA agarose bead than GFHM(HE)-IFN-${\gamma}$ with a 6X His-tag. rhIFN-${\gamma}$ was completely purified from enterokinase-treated HGFHM(HE)-IFN-${\gamma}$ by Ni-NTA affinity column. For high-level production of rhIFN-${\gamma}$, glucose was used as the sole carbon source with simple exponential feeding rate($2.4{\sim}7.2g/h$) in fed-batch process. The effective lactose concentration for the expression of the rhIFN-${\gamma}$ was $10{\sim}20mM$. Under the fed-batch culture conditions, rhIFN-${\gamma}$ production yield reached 11 g DCW/L for 6 hours after lactose induction.

Effect of Hot Water Extracts of Roasted Rhus vemiciflua Stokes on Antioxidant Activity and Cytotoxicity (볶음 처리한 옻나무 열수추출물의 항산화성 및 암세포 성장억제효과)

  • Kwak, Eun-Jung;Jo, Il-Jin;Sung, Ki-Seung;Ha, Tae-Youl
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.6
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    • pp.784-789
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    • 2005
  • The inhibitory effects of hot water extracts of Rhus verniciflua Stokes pith and peel roasted at 170, 200 and $220^{\circ}C$ on lipid peroxidation, formation of DPPH free radicals and growth of four human cancer cells such as HepG2 (liver cancer), SNU-1 (stomach cancer), MCF-7 (breast cancer) and Widr (colon cancer) were examined. The antioxidant activities and growth inhibitory effects on cancer cells of hot water extracts of peel were higher than those of pith, and the activities were dose-dependent. The roasting temperature showing the highest antioxidant activities and growth inhibitory effects on cancer cells was in the range of $170\~200^{\circ}C$ The lipid peroxidation and formation of DPPH free radicals of hot water extracts of roasted pith and peel were inhibited to 50.9, $56.5\%\;and\;79.0,\;78.4\%$ at the concentration of $500\mu g/mL$, respectively. The growth inhibitory effects of roasted pith and peel on cancer cells were in the order of Widr (41.5, $36.0\%$) > HepG2 (61.5, $44.0\%$) > MCF-7 (92.0, $69.2\%$)> SNU-1 (100, $100\%$) cells at the concentration of $1,000\mu g/mL$ as compared with the control, respectively. These results suggest that roasted Rhus verniciflua Stokes could be an useful natural medicinal plant for colon cancer.

Lipids and Free Sugar Composition in Ginseng Classified by Years (한국(韓國) 인삼(人蔘)의 연근별(年根別) 지질(脂質) 및 유리당조성(遊離糖組成))

  • Sohn, K.M.;Sung, T.S.;Cho, Y.J.;Lee, K.S.;Choi, C.
    • Applied Biological Chemistry
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    • v.31 no.2
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    • pp.169-176
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    • 1988
  • Lipid contents, fatty acid compositions and free sugar contents of dried ginsengs grown for different years were studied. The lipid and free sugar were separated and quantified by silicic acid column chromatography, thin layer chromatography, gas liquid chromatography and high performance liquid chromatography. Ginseng contained $1.07{\sim}67%$ crude lipid and the hightest lipid content was shown in 3 year old root. The lipid fractions obtained by silicic acid column chromatography were mainly composed of neutral lipid$(51.35{\sim}72.30%)$. The contents of phospholipid and glycolipid in total lipids were $15.03{\sim}34.59%$ and $11.83{\sim}20.72%$, respectively. As the ginseng grew, neutral lipid content increased gradually but glycolipid decreased to the half of the one year old root. Seven components of neutral lipid separated by thin layer chromatography were identified but the other two components were not able to be identified. Triglyceride content was $14.42{\sim}23.91%$, sterol ester and unidentified material(II) were $13.58{\sim}21.26%$ and $7.13{\sim}15.83%$ respectively. The major fatty acids were linoleic acid, palmitic acid, oleic acid and linolenic acid. Linoleic acid content of total and neutral lipid was $61{\sim}65%$ palmitic acid content in phospholipid was $26{\sim}39%$ and one year old root showed the hightest contents. The fatty acid composition of neutral lipid was similar to the pattern of total lipid. Linolenic acid content of glycolipid decreased gradually. Free sugars were composed of rhamnose, fructose, glucose, sucrose, maltose and an unidentified material. The content of sucrose was $92{\sim}94%$ of total free sugars. Two year old root showed the highest sugar content and it decreased since then.

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담자균류 ${\beta}$ -글루칸의 특성 및 생산

  • Hong, Eok-Gi
    • Proceedings of the Korean Society of Life Science Conference
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    • 2002.05a
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    • pp.17-22
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    • 2002
  • 지구상에는 수천종의 버섯류가 자생하고 있어 유전자원으로서의 중요성이 지대할 뿐만 아니라 기능성 식품소재 및 각종 약리 활성을 나타내는 신약개발 소재로도 크게 주목을 받고 있다. 이들 버섯은 균사체의 영양대사로 얻어지는 대사산물이 축적된 자실체의 형태로 나타나는데, 최근에 와서 자실체 및 균사체의 추출물이나 균사체 배양물이 체질개선이나 각종 병의 예방과 치료에 효과가 있는 것으로 밝혀져 건강식품이나 의약품으로서의 용도가 크게 증가하고 있는 실정이다. 특히, 담자균이 생산하는 특정 구조를 갖는 다당류는 오래전부터 종래의 화학요법제와는 달리 숙주내의 면역 기능을 부활하여 소위 면역요법제로서의 항암효과를 나타냄이 알려져왔었다. 현재까지 제약 및 의학적인 방법이 질병의 주된 치료방법으로 이용되어 왔지만 최근에 특정식품의 섭취가 만성질환의 발생을 억제 또는 지연시킨다는 연구 보고가 나오면서부터 만성질환의 치료방법으로서 식이요법을 중요하게 생각하게 되었다. 따라서 새로운 식품소재 및 가공식품의 개발을 통한 성인병 등의 각종 질병예방이 국민보건문제 해결에 필수적이다. 현재 일본 등에서는 표고버섯, 구름버섯 및 치마버섯 유래의 다당체 또는 단백다당체인 lentinan, krestin 또는 PS-K, schizophyllan 및 PSP 등이 실용화되어 높은 가격에 판매되고 있다. 국내에서도 야생 구름버섯 자실체로부터 추출한 단백 다당체인 Copolang(광동제약)이 개발되어 PS-K와 유사하게 암의 치료에 병행 사용되고 있고, 또 강력한 항암활성이 보고된 상황버섯의 균사체 추출물인 단백 다당체가 Mesima-Ex FK(한국신약)라는 상품명으로 암의 치료에 병행 사용되고 있는 것으로 알려지고 있다. 담자균류와 아울러 미생물 유래 다당체는 그 구조와 특성에 있어서 매우 다양함을 지니고 있다. 이러한 미생물 유래 다당류의 공업적 생산과 이용에 대한 연구로서는 Leuconostoc mesenteroides가 생산하는 dextran이 혈장증량제로 개발된 이래 Xanthomonas campestris가 생산하는 pullulan, Zoogloea rgmigera가 생산하는zooglan둥이 대표적인 예로 보고되고 있다. 한편, 미생물 유래 다당류는 구성당, 분자량, 화학적 구조 등과 같은 특성의 차이에 의해 많은 종류가 존재하고 있으며, 다양한 물성 및 유화제, 응고제, gel 형성제, 필름 형성제, 흡착제, 안정제, 접착제 등과 같은 용도로 광범위하게 이용되고 있다. 또한 근래에 들어서는 미생물 유래 다당체가 지니는 항암활성이 확인되어 새로운 의약품으로서의 개발 가능성이 기대된다. 그 밖에도 기존에 알려져 있는 식물 및 해조류 유래의 다당체와는 달리, 발효조를 이용한 연속배양에 의해 공업적 대량 생산이 가능하며, 더욱이 생산된 다당체의 분리 및 회수가 용이하다는 이점을 지니고 있다. 최근에 들어서는 유전공학적 기법을이용한 고생산성 변이균주 및 새로운 기능을 지닌 다당체의 개발에 관한 연구가 보고되고 있는 등 고부가가치를 지닌 새로운 바이오 소재로서의 기능 및 용도 개발에 관한 연구가 활발히 진행되고 있다. 이러한 항암 활성을 나타내는 여러 가지 담자균류중 Agaricus blazei로부터 생산되는 다당체는 고형암 이외에 S형 결장암, 난소암, 유방암, 폐암, 간암 등에 효과가 입증되었고, 천연물질에 의한 암 면역요법으로 각광을 받고 있으며, 항암 및 항virus의 완치율과 저지율에서 현재 여러가지 약효가 있는 버섯중에서도 탁원한 효과가 있는 것으로 증명되고 있다. 이들 다당체는 사이토카인을 생산시켜서 T임파구와 B임파구의 항원 특이적인 면역반응을 활성화시키고, 세포장해성 T세포와 활성화 대식세포의 세포장해 기능을 충진시켜서 암세포를 파괴시킨다. 또한 콜로니 자극인자인 사이토카인을 생산시켜서 면역담당세포의 신생을 촉진시키기도 하며, 암의 화학요법과 방사선 요법으로 저하된 백혈구를 회복시키는 역할을 한다. 따라서 최근의 연구동향은 생산된 다당체의 항암활성을 향상시키고자 하여 배양기간중에 interleukin을 의도적으로 첨가하는 경향이 있다. 이러한 항암 활성을 나타내는 담자균체 유래 다당체는 버섯의 기원에 따라 그 형태에 약간의 차이를 나타내기는 하나 그 기본 형태는 ${\beta}-(1,6)-glucosyl$ 분지를 가진 ${\beta}-(1,3)-glucan$이며, 평균 분자량은 50 ${\sim}$ 200만 정도이다. Agaricus blazei의 원산지인 브라질의 피에다데(Piedade) 지방의 환경조건(산지의 습도는 80%, 낮 기온 $35^{\circ)C$, 밤 기온 $20{\sim}25^{\circ}C$로 대단히 높으며, 정기적으로 열대지방 특유의 소나기가 내리는 지역)에서 볼 수 있듯이 Agaricus blazei의 성장 환경은 매우 까다로운 편이며, 날것으로는 보관이 잘 안되기 때문에 그 재배에 큰 어려움이 있다. 또한, 고체배양에 의해 생산된 버섯 자실체로부터 유기용매 및 열수추출 방법으로 다당체를 생산하는 방법은 균일한 형태의 버섯 자실체를 공급받기가 어렵기 때문에 다당체의 생산 수율이 낮고, 많은 노동력이 요구되는 어려움이 있다. 그러나 액체배양에 의한 다당체 생산의 경우는 고체배양에 의한 다당체 생산에 비해 일정한 조건하에서 배양이 가능하다는 장점이 있으며, 항상 균일한 균사체 및 배양액을 얻을 수 있다. 따라서 원하는 유용물질을 쉽게 획득할 수 있는 장점이 있다.

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Assessment of Nutritional Components, Carotenoid Content and Physiological Activity of Maize Hybrid for Grain 'Kangilok' (강일옥 옥수수의 영양성분, 카로티노이드 함량분석 및 생리활성 평가)

  • Lee, Ki Yeon;Kim, Jai Eun;Hong, Soo Young;Kim, Tae Hee;Park, A-Reum;Noh, Hee Sun;Kim, Si Chang;Park, Jong Yeol;Ahn, Mun Seob;Kim, Hee Yeon
    • Journal of Food Hygiene and Safety
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    • v.32 no.6
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    • pp.513-520
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    • 2017
  • This study was performed to provide basic data of 'Kangilok'. The objective of this study was to investigate worth of 4 parts of maize hybrid for grain, 'Kangilok' for functional foods. The 4 parts are kernels, dehulled kernels, skin of kernels and cobs of 'Kangilok'. We evaluated moisture, crude ash, crude lipid, crude protein, crude fiber and mineral content of 'Kangilok'. The moisture of kernels, dehulled kernels, skin of kernels and cobs of 'Kangilok' were 11.27%, 12.40%, 9.45%, 8.85% and the crude ash were 1.26%, 0.73%, 3.19%, 1.42%. Each of the crude lipid were 3.84%, 2.69%, 8.59%, 0.46% and the crude protein were 9.40%, 9.96%, 12.10%, 2.80%. The crude fiber of kernels, dehulled kernels, skin of kernels and cobs of 'Kangilok' were 2.24%, 0.92%, 7.07%, and 33.51%. Among the mineral contents, Ca and K content of cobs were highest by 4.84 mg/100 g, 114.33 mg/100 g and Fe, Mn contents of skin of kernels were highest by 5.30 mg/100 g, 2.64 mg/100 g. Mg content of kernels was the highest by 27.42 mg/100 g. P content of kernels, dehulled kernels, skin of kernels and cobs were 1.20%, 0.96%, 2.41%, and 0.19%. It was performed test on anti-oxidative, anti-inflammatory activities of 60% ethanol extract from 4 parts of Kangilok. The anti-oxidative effect was measured by DPPH and ABTS radical scavenging activity. As a results, DPPH radical scavenging activity (10 mg/mL) was 72.59%~93.05% and ABTS radical scavenging activity (10 mg/mL) was 48.17%~79.88%. The anti-inflammatory effect was measured by ability to inhibit production nitric oxide (NO) in RAW264.7 cell. As a result, all the extract of 4 parts were showed significantly inhibitory effect on NO production. Carotenoid contents quantified by using HPLC. ${\beta}$-Carotene of carotenoid was not analyzed in all the sample. Lutein and zeaxantin ware analyzed in kernels and skin of kernels.