• 제목/요약/키워드: P$N_2$(Purified $N_2$)

검색결과 471건 처리시간 0.025초

단백질 glycation 저해효과가 있는 식품소재 (Anti-glycation Activities from Various Agricultural Products)

  • 최희돈;최인욱;김윤숙
    • 한국식품과학회지
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    • 제39권4호
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    • pp.458-463
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    • 2007
  • 식품소재의 단백질 glycation 저해효과를 조사하기 위하여 곡류, 채소류 및 생약재 등 83종 에탄올 추출물에 대한 단백질 가교결합 저해효과를 $[^{14}C]$-N-formyl-lysine incorporation method를 이용하여 측정하였다. 대부분의 추출물에서 저해효과가 나타났으나 일부 추출물은 단백질의 가교를 촉진하는 것으로 나타났다. 상대적으로 높은 활성을 나타낸 시료 중 20개를 선별하여 형광광도법을 이용한 AGE형성 억제능을 조사한 결과, 1일 발아한 메밀의 추출물 (GB-l)이 가장 높은 저해활성을 가지는 것으로 나타났다. 차후 GB-0l은 콜로로포름, 에틸아세테이트, 부탄올 및 물로 분획되었으며 그 중 에틸아세테이트 분획이 가장 높은 단백질 glycation 저해활성(95.2% inhibitory activity at 100 ug/mL addi- tion level)을 보였다. HPLC를 이용한 폴리페놀 분석결과 발아 및 추출, 분획과정을 거친 메밀의 아세테이트 분획은 rutin 및 quercetin 함량이 상당히 증가한 것으로 나타나서 부분정제물의 rutin 및 quercetin 함량과 저해활성사이에 높은 상관성을 나타내었다.

DNA에 결합하는 항암제의 작용기전 (Mechanism of Action of Anticancer Drug Aziridinylbenzoquinones: Involvement of DT-diaphorase)

  • Lee, Chong-Soon-
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1994년도 제2회 추계심포지움
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    • pp.147-172
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    • 1994
  • Aziridinylbenzoquinones such as 3, 6-diaziridinyl-1, 4-benzoquinone (DZQ) and its 2, 5-methyl analog (MeDZQ) require bioreductive activation in order to elicit their anticancer activities. To determine the involvement of DTD in the activation of these drugs, we have used a ligation-mediated polymerase chain reaction to map the intracellular alkylation sites in a sing1e copy gene at the nucleotide level. We have performed this analysis in two human colon carcinoma cells, one proficient (HT-29) and one deficient (BE) in DT-diaphorase (DTD) activity. In the DTD proficient HT-29 cell line, DZQ and MeDZQ were found to alkylate both 5'-(A/T)G(C)-3' and 5'-(A/T)A-3' sequences. This is consistent with the nucleotide preferences observed when DZQ and MeDZQ are activated by purified DTD to reactive metabolites capable of alkylating DNA in vitro [Lee, C. -S., Hartley, J. A., Berardini, M. D., Butler, J., Siegel., D., Ross, D., & Gibson, N. W. (1992) Biochemistry, 31: 3019-3025]. Surprisingly in the DTD-deficient BE cell line a pattern of alkylation induced by DZQ and MeDZQ similar to that observed in the DTD-proficient HT-29 cells was observed. This suggests that reductive enzymes other than DTD can be involved in activating DZQ and MeDZQ to DNA reactive species in vivo.

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Purification and Characterization of Chitinase from a Marine Bacterium, Vibrio sp. 98CJ11027

  • 박신혜;이정현;이홍금
    • 미생물학회지
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    • 제38권4호
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    • pp.224-224
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    • 2002
  • Chitin-degrading marine bacterial strain 98CJ11027 was isolated from bryozoa from the coastal area of Cheju Island, Korea, and identified as a member of the genus Vibrio. The molecular mass of the main extracellular chitinase (chitinase I), purified from strain 98CJ11027, was estimated to be 98 kDa. The optimal condition for chitinase I activity is pH 6.0 and 45℃. The activity was inhibited by $Fe^+2$ and$Cu^+2$. Chitinase I displayed the hydrolysis type of chitobiosidase and catalyzed reversed hydrolysis leading to the synthesis of tetraacetylchitotetraose.

Antibacterial Activity of Oleanolic Acid from Physalis angulata against Oral Pathogens

  • Hwang, Jae-Kwan;Shim, Jae-Seok;Park, Kyung-Min;Chung, Jae-Youn
    • Preventive Nutrition and Food Science
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    • 제7권2호
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    • pp.215-218
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    • 2002
  • A methanol extract of Physalis angulata exhibited in vitro antibarterial activity against oral pathogens such including Streptococcus mutans and Porphyromonas gingivalis. The methanol extract of Physalis angulata was further fractionated with ethyl acetate, n-butanol and water, in which the ethyl acetate fraction exclusively showed antibacterial activity. An active antibacterial compound from the ethyl acetate fraction was purified to a single compound using silica gel column chromatography and identified as oleanolic acid by $^{13}$ C-NMR, $^1$H-NMR and EI-MS. MIC of oleanolic acid against S. mutants and p. gingivalis were determined to be 50 and 25 ug/mL, respectively. The Antibacterial activity of oleanolic acid from Physalis angulata suggested that it has potential as an anticarcinogenic and antiperiodontic ingredients in various foods and oral care products.

Co-expression of Gamma-Aminobutyrate Aminotransferase and Succinic Semialdehyde Dehydrogenase Genes for the Enzymatic Analysis of Gamma-Aminobutyric Acid in Escherichia Coli

  • So, Jai-Hyun;Lim, Yu-Mi;Kim, Sang-Jun;Kim, Hyun-Ho;Rhee, In-Koo
    • Journal of Applied Biological Chemistry
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    • 제56권2호
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    • pp.89-93
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    • 2013
  • Gamma-aminobutyric acid (GABA) aminotransferase (gabT) and succinic semialdehyde dehydrogenase (gabD) genes from Pseudomonas fluorescens KCCM 12537 were cloned into a single pETDuet-1 vector and co-expressed in Escherichia coli BL21(DE3) simultaneously. The mixture of both enzymes, called GABase, is the key enzyme for the enzymatic analysis of GABA. The molecular mass of the GABA aminotransferase and succinic semialdehyde dehydrogenase were determined to be 52.8 and 46.7 kDa following computations performed with the pI/Mw program, respectively. The GABase activity between pH 6.0 and 9.0 for 24 h at $4^{\circ}C$ remained over 75%, but under pH 6.0 decreased rapidly. The GABase activity between 25 and $35^{\circ}C$ by the treatment at pH 8.6 for 30 min remained over 80%, but over $35^{\circ}C$ decreased rapidly. When the activity against GABA was defined as 100%, the purified GABase activity against 5-aminovaleric acid having a similar structure to GABA showed 47.7% and GABase activity against ${\beta}$-alanine, ${\varepsilon}$-amino-n-caproic acid, $_L$-ornithine, $_L$-lysine, and $_L$-aspartic acid showed between 0.3 to 2.3%. The GABA content was analyzed with this co-expressed GABase, compared with the other GABase which was available commercially. As a result, the content of GABA extracted from brown rice, dark brown rice, and black rice were $26.4{\pm}3.5$, $40.5{\pm}4.7$ and $94.7{\pm}9.3{\mu}g/g$, which were similar data of other GABase in the error ranges.

황기 종자의 천연 항진균성 단백질의 분리정제 및 특성검정 (Purification and Characterization of Natural Antifungal Protein from Astragal Seeds (Astragalus membranaceus L.).)

  • 구본성;류진창;정태영;김교창
    • 한국미생물·생명공학회지
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    • 제26권5호
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    • pp.379-386
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    • 1998
  • 본 연구에서는 천연 항균물질의 개발 이용을 위해 황기 종자로부터 인체에 무해한 천연 항균 단백질을 ion exchange chromatography 및 gel filteration을 이용하여, 순수 분리하고 특성을 조사하였다. 황기종자로부터 추출한 천연 항균 단백질은 Aspergillus ocraceus, Penicillium expensum, P. digitatum, Botrytis cineria의 포자 발아 및 효모인 Candida albicans의 생육을 현저하게 저해하였으며 ammonium sulfate 포화도가 0.4일 때 단백질의 침전량이 122.6 $\mu\textrm{g}$/$m\ell$로 가장 많았고 항균력도 15.2 mm로 가장 높게 나타났다. 강력한 cation exchange chromatography인 Mono-S를 이용하여 FPLC에서 단백질을 분획하였을때 첫번째 peak에서 분획된 단백질군이 항균력을 보였으며 Superose 12HR gel filteration column을 이용하여 2차 분획 하였을 때 분자량이 19 kDa되는 단일 단백질만을 순수분리 할 수 있었다. 전기 영동한 polyacrylamide gel위에 곰팡이 포자를 중층하는 bio autography로 19 kDa 단백질 band의 항균력을 직접 확인하였으며 분리된 항균 단백질의 아미노 말단의 아미 노산 22잔기를 sequencing하고 thaumatin 및 zeamatin 유사 단백질들과 상동성을 측정한 결과 50%내외의 homology를 나타내었다. 분리된 항균 단백질은 곰팡이 균사가 성장하는 선단부위에 가장 먼저 침투하여 channel을 형성함으로 osmolysis를 일으켜 곰팡이의 생육을 억제하는 것으로 추측할 수 있었다.

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Investigation of Siderophore production and Antifungal activity against Phytophthora capsici as related to Iron (III) nutrition by Lysobacter antibioticus HS124

  • Ko, Hyun-Sun;Tindwa, Hamisi;Jin, Rong De;Lee, Yong-Seong;Hong, Seong-Hyun;Hyun, Hae-Nam;Nam, Yi;Kim, Kil-Yong
    • 한국토양비료학회지
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    • 제44권4호
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    • pp.650-656
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    • 2011
  • Lysobacter antibioticus HS124 isolated from pepper rhizosphere soil produced catechol type siderophore. Purified siderophore by Diaion HP-20 and silica gel column chromatography showed several hydroxyl functional groups adjacent to benzene rings by analysis of $^1H$ NMR spectroscopy. The strain HS124 showed different activities to suppress Phytophthora capsici with different concentrations of exogenous Fe (III) in minimal medium where antifungal activity with $100{\mu}M$ Fe (III) was approximately 1.5 times higher than in absence of Fe (III). Bacterial population in this Fe (III)-amended medium was also highest with $8.9{\times}10^8\;CFU\;ml^{-1}$ which also corresponded to the strongest siderophore activity. When grown in rich medium (minimal medium with N, $P_2O_5K_2O$ and glucose), HS124 exhibited approximately 2 times stronger antifungal activity compared to minimal medium. In pot trials, treatments of bacterial culture grown in rich medium with (C1) or without (C2) $100{\mu}M$ Fe (III) exhibited a high protection of pepper plants from disease, compared to medium only with (M1) or without (M2) $100{\mu}M$ Fe (III). Especially, treatment C1 showed the best disease control effect of about 70 %. Thus, the strain HS124 should be recommended as a potential biocontrol agent against P. capsici in pepper.

광합성세균 Rhodopseudomonas gelatinosa 의 시토크롬 c 산화효소의 정제 및 특성 (Purification and Characterization of Cytochrome c Oxidase from Photosynthetic Bacterium, Rhodopseudomonas gelatinosa)

  • 강대길;최원기
    • 미생물학회지
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    • 제30권2호
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    • pp.101-107
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    • 1992
  • 화학 영양성으로 배양한 Rps. gelatinosa 에서 2회의 시토크롬 c 친화성 크로마토그래피와 DEAE-Sephacel 이온 교환 크로마토그래피 등 3 단계의 크로마토그래피를 수행하여 시토로콤 c 산화효소를 정제하였다. 정제된 시토크롬 c 산화효소는 Sephacryl S-300 에 의한 분자걍이 약 110,000 Da 이고 SDS-gel 전기영동에 의한 분자량이 약 52.000 Da 으로써 이량체일 것으로 보인다. 전제된 시토크롬 c 산화효소는 온도데 매우 불안정하고 말 심장 시토크롬 c 를 기질로 사용했을때 Km 값은 $20\mu$M, Vmax 값은 44unit/mg prot. 이며 pH 6.4 의 효소방응 최적 pH 와 25.deg.C 의 최적 온도를 보였다. 환원된 시토크롬 c 산화효소는 554, 523, 421 nm 에서 .alpha., .betha. soret 흡수대를 보였고 chromatophore 에서와 마찬가지로 KCN 과 $NaN_{3}$ 에 의해서는 효소 활성도가 저해를 받았지만 CO 와 antimycin A, myxothiazol 에 의해서는 효소 활성도가 저해를 받지 않았다. 빛을 에너지원으로 배양하거나 또는 화학영양성으로 배양하든지 모두 시토크롬 c-551 이 생성되었고 환원된 시토크롬 c-551 은 시토크롬 c 산화효소에 의해 산화되었다. 시토크롬 c-551 을 기질고 이용하였을 때 시토크롬 c 산화효소의 Km 값은 $26\mu$M 이었고 Vmax 값은 31.unit./mg prot. 로써 말심장의 시토크롬 c 를 기질로 이용할때 보다 오히려 낮았다. 이와 같은 결과로 보아 화학 영양성은 배양한 Rhodopseudomonas gelatinosa 에서 호흡에 의한 전자전달은 시토크롬 c-551 이 시토크롬 $bc_{1}$ 복합체로 부터 전자를 받아 b-형 시토크롬 c 산화효소에 전자를 전달해 주고 최정적으로 산소를 환원시킬 것으로 생각된다.

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Effects of Dietary Arachidonic Acid (20:4n-6) Levels on Growth Performance and Fatty Acid Composition of Juvenile Eel, Anguilla japonica

  • Bae, Jun-Young;Kim, Dae-Jung;Yoo, Kwang-Yeol;Kim, Sun-Gyu;Lee, Jeong-Yeol;Bai, Sungchul C.
    • Asian-Australasian Journal of Animal Sciences
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    • 제23권4호
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    • pp.508-514
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    • 2010
  • This study was conducted to evaluate the effects of dietary arachidonic acid (AA, 20:4n-6) levels on growth performance and body composition in juvenile eel, Anguilla japonica. Six semi-purified experimental diets were formulated to be isonitrogenous and iso-caloric containing 55.0% crude protein and 15% crude lipid (18.3 kJ of available energy $g^{-1}$). Six different levels of AA were added to the basal diet, with 0, 0.2, 0.4, 0.6, 0.8 or 1.2% on a dry matter (DM) basis, respectively ($AA_{0.07},\;AA_{0.22},\;AA_{0.43},\;AA_{0.57},\;AA_{0.78}\;or\;AA_{1.23}$). After a conditioning period, fish initially averaging 27${\pm}$0.5 g (mean${\pm}$SD) were randomly distributed into each aquarium as triplicate groups of 20 fish each. One of six experimental diets was fed on a DM basis to fish in three randomly selected aquaria at a rate of 2-3% of total body weight twice a day. At the end of the 12-week feeding trial, weight gain (WG) and feed efficiency (FE) of fish fed $AA_{0.78}$ and $AA_{1.23}$ diets were significantly higher than of fish fed $AA_{0.07},\;AA_{0.22},\;AA_{0.43}$ diets (p<0.05). Specific growth rate (SGR) of fish fed the $AA_{0.78}$ diet was significantly higher than of fish fed $AA_{0.07},\;AA_{0.22},\;AA_{0.43}$ diets (p<0.05). However, there were no significant differences in WG, SGR and FE among fish fed $AA_{0.57},\;AA_{0.78}\;or\;AA_{1.23}$ diets (p>0.05). Whole body AA deposition of fish fed the $AA_{1.23}$ diet was significantly higher than for the other diets (p<0.05). Broken-line model analysis on the basis of WG and SGR indicated that the dietary AA requirement could be greater than 0.69% but less than 0.71% of the diet in juvenile eel. The growth-promoting activity of AA observed in the present study provides strong support for the contention that dietary AA is essential for juvenile eel.

Cellulosimicrobium sp. YB-43으로부터 mannanase C 유전자의 클로닝과 효소 특성 (Gene cloning of β-mannanase C from Cellulosimicrobium sp. YB-43 and characterization of the enzyme)

  • 윤기홍
    • 미생물학회지
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    • 제54권2호
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    • pp.126-135
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    • 2018
  • 여러 종류의 mannanase를 생산하는 Cellulosimicrobium sp. YB-43으로부터 mannanase B를 암호하는 manB 유전자와 효소의 특성이 보고된 바 있다. Mannanase C (ManC)로 명명한 효소의 유전자가 manB 유전자의 하류에 위치한 것으로 예상되어 이를 중합효소 연쇄반응으로 클로닝하여 manC 유전자의 염기서열을 결정하였다. ManC는 448 아미노산 잔기로 구성된 것으로 확인되었으며 glycosyl hydrolase family 5에 속하는 mannanase와 상동성이 높은 활성영역과 탄수화물 결합영역(CBM2)이 존재하였다. ManC의 활성영역은 Streptomyces sp. SirexAA-E (55.8%; 4FK9_A) 및 S. thermoluteus (57.6%; BAM62868)의 mannanase와 아미노산 배열의 상동성이 55% 이상으로 가장 높았다. Signal peptide 영역이 제거되고 카르복실 말단에 hexahistidine이 연결되도록 제조한 His-tagged ManC (HtManC)의 유전자를 재조합 대장균에서 발현하여 균체 파쇄액으로부터 HtManC를 정제하였다. HtManC은 $65^{\circ}C$와 pH 7.5에서 최대 활성을 보였으며 pH 7.5~10범위에서 활성에 큰 변화가 없었다. HtManC는 locust bean gum (LBG)과 konjac에 대한 분해 활성이 guar gum과 ivory nut mannan (ivory nut)에 비해 높았다. 최적 반응조건에서 LBG를 기질로 하여 반응 동력학적 계수를 측정한 결과 Vmax와 Km이 68 U/mg과 0.45 mg/ml로 나타났다. HtManC에 의한 만노올리고당(MOS)과 mannan의 분해산물을 TLC로 관찰한 결과 mannobiose 보다 중합도가 큰MOS로부터 mannobiose와 mannotriose가 주된 분해산물로 생성되었다. 또한 LBG, konjac과 ivory nut의 분해산물로 mannobiose와 소량이 mannose가 공통적으로 관찰되었다.