• Title/Summary/Keyword: 인산분해효소

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Anti-aging Effect of Akebia quinata Decaisne Ethanol Extract (으름덩굴 에탄올 추출물의 항노화 효과)

  • Yu Jin Kim;Soon Hyun Kwon;Ji Hyun Song;So Mi Lee;Yong Min Kim
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.50 no.1
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    • pp.67-75
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    • 2024
  • Skin aging progresses due to external factors such as ultraviolet rays and infections. These factors cause skin fibroblasts to secrete proteolytic enzymes, matrix metalloproteinases (MMPs). MMPs induce the degradation of collagen located in the extracellular matrix, directly influencing aging. The stems of Akebia quinata Decaisne have been reported to have antioxidant and anti-inflammatory effects. However, the anti-aging effect of Akebia quinata Decaisne stem ethanol extract (AQSEE) is not known. Therefore, we studied the TNF-α-induced MMP-1 inhibitory effect in human fibroblasts. When the cell viability of AQSEE was confirmed through MTT asaay, it showed no toxicity up to 400 ㎍/mL. The inhibition of MMP-1 mRNA and protein secretion was confirmed through RT-qPCR and ELISA, and results showed a significant decrease at concentrations of 100, 200, 400 ㎍/mL. We also confirmed by Western blotting that phosphorylation of MAPKs signaling pathway and transcription factors was reduced. As a result, phosphorylation of p38, c-Jun, p65 was significantly decreased at all concentrations. DPPH and ABTS assays were performed to confirm the radical scavenging ability of AQSEE, and the results showed a significant decrease at all concentrations. The results of this study confirmed the MMP-1 inhibitory effect and radical scavenging ability, which suggests that it can be used as an anti-aging substance.

Measurement of Biogenic Amines with a Chitopearl Enzyme Reactor (Chitopearl 효소 Reactor를 이용한 Biogenic Amines 측정)

  • Park, In-Seon;Kim, Dong-Kyung;Shon, Dong-Hwa;Cho, Yong-Jin;Kim, Nam-Soo
    • Korean Journal of Food Science and Technology
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    • v.31 no.3
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    • pp.593-599
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    • 1999
  • Substrate specificity of a flow-injection-analysis (FIA)-type biogenic amine sensor with enzyme reactor was determined. The enzyme reactor was prepared with a diamine oxidase immobilized on preactivated chitosan porous beads (Chitopearl) by intramolecular cross-linking via glutaraldehyde. The sensor showed a rapid response to putrescine and a quasi-linear calibration curve was obtained up to 15.0 mM. The optimal pH and temperature of the enzyme reactor system were 7.5 and $35^{\circ}C$. Interferences due to ATP-related compounds and trimethylamine, and the effects of NaCl and amino acids were measured. Inhibitory effects owing to these components could be mitigated by sample extraction with perchloric acid. Polyamines except putrescine were determined by a putrescine calibration range within 26.7%. This system was confirmed as rapid and convenient for biogenic amine determination.

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Biological Activities of Calcium Polyphosphate (Calcium polyphosphate의 생물학적 활성도에 관한 연구)

  • Seol, Yang-Jo;Lee, Jae-Il;Lee, Yong-Moo;Lim, Yoon-Tak;Kim, Seok-Young;Ku, Young;Rhyu, In-Chul;Hahm, Byung-Do;Han, Soo-Boo;Choi, Sang-Mook;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.30 no.2
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    • pp.213-231
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    • 2000
  • 이 연구의 목적은 다공성의 CPP 내부에 쥐의 장골의 골수에서 유래된 세포를 접종하고 3차 원적으로 배양하여 CPP가 골 형성을 위한 조직공학의 지지체로 적용가능한가를 연구하는 것과 Calcium PolyPhosphate(CPP)의 돌연변이 유발성을 검사하는 것이다. 무수 ($Ca(H_2PO_4)$)를 condensation하여 무결정의 ($Ca(PO_3)$)를 얻고 이를 용융하고 냉각시킨 후 분쇄하여 Calcium polyphosphate(CPP) powder를 얻었다. 다공성의 CPP는 5% $SiO_2$를 첨가하여 sponge 형태로 $450-550{\mu}m$ 소공의 크기를 가지는 것과(CPP-45ppi) $200-300{\mu}m$의 소공의 크기를 가지는 것(CCP-60ppi) 2가지로 제작하였다. 각각의 CPP matrices는 $5mm{\times}5mm{\times}1mm$의 블록 형태로 만들었다. 체중 100g 내외의 백서에서 장골(femur, tibia)을 채취하여 백서의 장골 골수 세포를 분리하여 배양한 후 24well에 CPP block을 넣고 CPP block 당 $10^5$개의 배양한 세포를 접종하였다. 배양 1, 7, 14, 및 21 일째에 각 well에서 trypsin EDTA를 이용하여 2회 반복하여 cell을 분리하였고, 원심분리한 후 hemacytometer로 측정하였다. 또, 45ppi와 60ppi, 그 리 고 Tissue Culture Polystyrene(control group)에 접종, 배양된 세포들의 염기성 인산분해효소활성도를 배양 7, 14, 및 21 일째에 각각 측정하였다. 각 기간별로 배양된 세포-CPP 혼합체내에서 세포의 부착 및 증식과 형성된 조직의 3차원적 형태를 관찰하기 위하여 주사전자현미경하에서의 관찰하였다. CPP의 돌연변이 유발성 검사 (mutagenicity test)를 위해 hypoxanthine-guanine phosphoribosyl transferase(HPRT) assay를 하였다. NIH3T3 cell line과 CHO-K1 cell line으로 각각 $1000{\mu}g/m{\ell}$, $100{\mu}g/m{\ell}$, $10{\mu}g/m{\ell}$ 그리고 $1{\mu}g/m{\ell}$의 CPP 농도에서 측정하였다. 통계적 분석을 위해서 모든 측정은 각군당 4개체 이상 시험하였고, 각 측정값은 평균값${\pm}$표준편차로 나타내었다. 각 군간의 통계적 유의성 검정을 위해서 Analysis of variance(ANOVA)를 이용하였고 Tukey의 방법으로 사후분석을 실시하였다. 제작된 CPP matrices 소공들이 서로간에 연결이 잘 되어있는 형태였다. 두 가지로 제조된 CPP(45ppi와 60ppi) 모두에서 세포의 부착이 잘 일어났고, 부착된 세포의 분열도 잘 일어났다. 2 가지의 CPP 모두에서 7, 14, 21일째의 세포 수는 1일째에 비해 유의성 있게 증가하였다(P<0.01). 3차원적 구조인 Calcium PolyPhosphate에서 배양한 세포는 24well dish(tissue culture polystyrene)에서 평면적으로 배양한 대조군의 세포에서 보다 염기성 인산분해효소 (Alkaline Phosphatase)를 유의성 있게 높게 나타냈다. 주사전자현미경에서 세포-CPP 혼합체를 관찰한 결과, CPP block에 세포들이 잘부착되어 있었고, 시간이 지남에 따라 세포가 여러 층을 형성하면서 뭉치는 현상을 보였다. 또, HPRT assay 결과 , Calcium PolyPhosphate는 돌연변이 유발성을 보이지 않았다. 이상의 결과로 볼 때 CPP에는 세포부착이 잘 일어나고, 지지체 상에서 세포의 분열도 활발하게 일어나므로 골조직을 위한 조직공학의 우수한 지지체가 될 수 있을 것으로 사료된다.

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Isolation and Characteristics of Bacteria Showing Biocontrol and Biofertilizing Activities (생물방제 및 생물비료 활성을 가지는 세균의 분리 및 특성)

  • Jung, Ho-Il;Kim, Keun-Ki;Park, Hyean-Cheal;Lee, Sang-Mong;Kim, Yong-Gyun;Kim, Hong-Sung;Lee, Cnung-Yeol;Son, Hong-Joo
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1682-1688
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    • 2007
  • To develop multifunctional microbial inoculant, microorganisms with antagonistic activity and biofertilizing activity were screened. Pantoea agglomerans and Bacillus megaterium from our laboratory culture collection, and strain MF12 from soil near poultry farm in Miryang were selected. On the basis of morphological, physiological studies and 16S rDNA sequence analysis, isolate MF12 was identified as the Bacillus pumilis. Three strains were studied for insoluble phosphate solubilization, indole-3-acetic acid (IAA) and siderophore production, ammonification ability, hydrolytic enzyme production and antifungal activity against phytopathogenic fungi. P. agglomerans did not produce any visible clear zone on agar plate containing 0.5% $Ca_3(PO_4){_2}$ as a sole phosphorus source. However, this strain could solubilize insoluble phosphate in liquid medium. All strains produced IAA ranged from $3{\sim}639{\mu}g/ml$ depending on culture time and had ammonification ability. Among three strains, only P. agglomerans produced siderophore. P. agglomerans produced pectinase and lipase, B. megaterium produced amylase, protease and lipase while B. pumilis produced protease and lipase. P. agglomerans showed antifungal activities against phytopathogenic fungi, Fusarium oxysporum and Colletotrichum gloeosporioides. B. pumilis showed antifungal activities against Botrytis cinerea, Sclerotinia sclerotiorum and Phythium ultimum.

Production of a Phospholipase C by Bacillus cereus and Its Characterization (Bacillus cereus에 의한 C형 인지질 분해효소의 생산과 특성 고찰)

  • 서국화;임용식;이종일
    • KSBB Journal
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    • v.19 no.4
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    • pp.250-256
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    • 2004
  • In this work we have cultivated several B. cereus strains in a complex LB medium in order to study the production of phospholipase C (PLC), and among them B. cereus 318 showed the highest productivity of PLC. Some components, i.e., 5 g/L glucose, 5 g/L yeast extract, 5 g/L peptone, 0.5∼1.0 g/L K$_2$HPO$_4$, 0.02∼0.04 g/L ZnSO$_4$$.$7H$_2$O and 3 g/L NaHCO$_3$ were found to be optimal for the high production of PLC by B. cereus 318. Optimal culture temperature and pH were found to be 30$^{\circ}C$ and pH 7.5 for the PLC production, respectively. Optimum reaction temperature and pH of the PLC produced by B. cereus 11 and 318 were 45$^{\circ}C$ and pH 4.0, while they were 50$^{\circ}C$ and pH 7.0 for the PLC by B. cereus 559. The PLC produced by B. cereus was activated by Mn$\^$2+/, Co$\^$2+/ and dimethyl sulfoxide (DMSO), but its activity was inhibited by Cu$\^$2+/ and partially by glycerol, isopropanol and sodium dodecyl sulfate (SDS).

Antibacterial Activity of Sodium Phytate Against Salmonella typhimurium in Meats (식육에서 피틴산염의 Salmonella typhimurium균에 대한 항균효과)

  • Baek, Dong-Jin;Hue, Jin-Joo;Lee, Yea-Eun;Lee, Ki-Nam;Nam, Sang-Yoon;Yun, Young-Won;Jeong, Jae-Hwang;Lee, Sang-Hwa;Lee, Beom-Jun
    • Journal of Food Hygiene and Safety
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    • v.22 no.4
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    • pp.382-387
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    • 2007
  • The approval of use of certain food-grade phosphates as food additives in a wide variety of meat products greatly stimulated research on the applications of phosphates in foods. Phytic acid is a natural plant inositol hexaphosphate constituting 1-5% of most cereals, nuts, legumes, oil seeds, pollen, and spores. In this study, we investigated antibacterial activity of sodium phytate (SPT) against Salmonella typhimurium in tryptic soy broth with different pHs and in chicken, pork and beef. In tryptic soy broth, SPT at the concentrations of 0.1, 0.5, and 1.0% effectively inhibited the growth of Salmonella typhymurium in a concentration-dependent manner. At pH 5.5-7.0 similar to meat pHs, 1% SPT almost completely inhibited the bacterial growth. The inhibitory effect of SPT was stronger at pH 7.0 than pH 5.5. In chicken, pork, and beef, SPT at the concentrations of 0.1, 0.5, and 1% significantly inhibited the growth of Salmonella typhimurium in a dose-dependant manner (p<0.01). The addition of 1% SPT in the meats significantly increased the meat pHs. These results indicate that SPT is very effective for inhibition of bacterial growth as a muscle food additive for increasing food safety and functions.

Secondary Metabolites from Enzymatic Oxidation of Caffeic Acid with Pancreatic Lipase Inhibitory Activity (카페인산의 효소적 산화반응으로부터 췌장 지방분해효소 저해 물질의 분리)

  • Kim, Tae Hoon;Kim, Myoung Kwon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.12
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    • pp.1912-1917
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    • 2015
  • Pancreatic lipase is a potential therapeutic target for the treatment of diet-induced obesity in humans. As part of our continuing search for novel bioactive compounds, the convenient enzymatic transformation of caffeic acid into neolignans as well as related oxidized-products enhanced pancreatic lipase inhibitory activity. Enzymatic transformation of caffeic acid (1) using polyphenol oxidase originating from Korean pear yielded four oxidized metabolites, which were identified by different spectroscopic techniques ($^1H$,$^{13}C$ NMR, DEP/T, $^1H-^1H$ COSY, HMBC, HMQC, and NOESY). The anti-obesity efficacy of caffeic acid reactant was tested by in vitro porcine pancreatic lipase assay. All tested samples showed dose-dependent pancreatic lipase inhibitory activities. Four oxidative products including phellinsin A (2), caffeicinic acid (3), isocaffeicinic acid (4), and 7,8-erythro-caffeicin (5) were isolated and identified. The major metabolites (2~5) were evaluated for their pancreatic lipase inhibitory activity, and oxidized-products (2~3) improved potency against pancreatic lipase when compared to original caffeic acid. This result suggested that the neolignans isolated from oxidative transformation of caffeic acid might be beneficial in the treatment of obesity and relevant diseases, and the convenient enzymatic transformation by polyphenol oxidase may be a valuable method for structural modification and enhancement of activity.

The Effect of the Use of Concept Mapping on Science Achievement and the Scientific Attitude in Ocean Units of Earth Science (해양단원 개념도 활용 수업이 과학성취도 및 태도에 미치는 효과)

  • Han, Jung-Hwa;Kim, Kwang-Hui;Park, Soo-Kyong
    • Journal of the Korean earth science society
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    • v.23 no.6
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    • pp.461-473
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    • 2002
  • Concept mapping is a device for representing the conceptual structure of a subject discipline in a two dimensional form which is analogous to a road map. In the teaching and learning of earth science, each concept depends on its relationships to many others for meaning. Using concept mapping in teaching helps teachers and students to be more aware of the key concepts and relationships among them. The purpose of this study is to investigate the effect of the use of concept mapping on science achievement and the scientific attitude in ocean units of earth science. The results of this study are as follows; first, the science achievement of a group of concept mapping teaching is significantly higher than that of the group of traditional teaching. Also, when the achievement levels are compared among different cognitive ability groups, the effect is more significant in mid or lower level student groups than in high level groups. The use of concept mapping is more effective when the concepts have a distinct concept hierarchy. Second, the scores of the test of ‘attitude toward scientific inquiry’ and ‘application of scientific attitude’ of the group of concept mapping teaching are significantly higher than those of the group of traditional teaching, whereas the scores of the test of ‘interest in science learning’ of concept mapping teaching is not different from those of group of traditional teaching. Third, the survey on the use of concept mapping shows a positive response across the tested groups. The use of concept mapping is more beneficial in fostering the comprehension of the topic. A concept map of student's own construction facilitates the assessment of learning, thus promising the usefulness of concept mapping as a means of evaluation. In regard to retention aspect, concept mapping is considered to be more effective in confirming and remembering the topic, while less effective in the aspects of activity and interest. In conclusion, the use of concept maps makes learning an active meaningful process and improves student's academic achievement and scientific attitude. If the concept mapping is more effectively as an active teaching strategy, more meaningful learning will be attained.

Effects of Several Natural Medicines on Alkaline Phosphatase Activity in hFOB1 (수종의 생약제제가 hFOB1의 염기성 인산분해 효소 활성에 미치는 영향)

  • Jang Kil Young;Hyun Ha Na;Kim Yun Sang;You Hyung Keun;Shin Hyung Shik
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.5
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    • pp.1042-1047
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    • 2002
  • Recently, many natural medicines, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, anti-inflammatory and regenerative potential for periodontal tissues. Cortex Eucommiae, Eupoly phaga, Semen Cuscutae, Halloysitum Rubrum have been traditionally used as medicines for treatment of bone disease in Korea. The objective of the present study is to examine the ability of alkaline phosphatase (ALP) activity in human fetal osteoblast cell line (hFOB1) with several natural medicines. hFOB1 added DMEM/F-12 were cultured with dexamethasone as a positive control, and with each natural medicine. ALP activity was measured by spectrophotometer for enzyme activity and naphthol AS-Bl staining was performed for morphometry. All of the natural medicines induced a higher ALP activity compared to negative control, especially, Cortex Eucommiae increased an ALP activity in all experimental groups (p<0.05). In naphthol AS-Bl staining, all of the natural medicines of this study increased the stained area compared to negative control. Especially, Cortex Eucommiae and Eupoly phaga showed statistical significance compared to negative control (p<0.05). These results indicate that Cortex Eucommiae, Eupoly phaga, Semen Cuscutae, Halloysitum Rubrum have an inducing ability of ALP synthesis on osteoblasts.

Effects of Several Herbal Medicines on Alkaline Phosphatase Activity in Human Fetal Osteoblasts (수종의 생약제제가 human fetal osteoblasts의 염기성 인산분해 효소 활성에 미치는 영향)

  • Lee, Myoung-Ku;Choi, Hee-In;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.33 no.1
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    • pp.49-60
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    • 2003
  • Several growth factors and polypeptides are not commonly yet used for regenerators of bone tissue or alveolar bone because of the insufficiency of studies on their side effects, genetic engineering for mass production and stability for clinical application. Recently, many herbal medicines, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, antiinflammatory and regenerative potential of periodontal tissues. Morindae Radix, Cibotium Barometz (L.), Albizziae Cortex, Cistandhis Herba have been traditionally used as medicines for treatment of bone disease in Eastern medicine. The objective of the present study is to examine the ability of alkaline phosphatase (ALP) activity of human fetal osteoblast (hFOB1) when several natural medicines were supplemented. hFOB1 were cultured with Dulbecuo's Modified Eagle's Medium Nutrient Mixture F-12 HAM ( DMEM/F-12 1:1 Mixture, Sigma, USA) and negative control, dexamethasone (positive control), and each natural medicines for 3 days. And then ALP activity was measured by spectrophotometer for enzyme activity and Alizarin red S staining for morphometry. Among the natural medicines of this study, Morindae Radix, Cibotium Barometz (L.) and Cistanchis Herba induced higher activity of ALP synthesis than negative controls in all experimental group. Albizziae Cortex showed mild increases than negative control group. According to measurement of positively stained area, all of the natural medicines of this study increased compared to negative control. Especially, Cibotium Barometz (L.) and Cistanchis Herba showed statistical significance compared to negative control (p<0.05). These results indicate that Morindae Radix, Cibotium Barometz (L.), Albizziae Cortex, Cistandhis Herba have an inducing ability of ALP synthesis on osteoblast.