• 제목/요약/키워드: Production and Purification

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유채 전초 추출물 유래 항산화 및 항염 활성 성분 (Anti-oxidative and Anti-inflammatory Constituents from the Extracts of Brassica napus L. Whole Plant)

  • 조연정;현주미;강지미;김창윤;이남호
    • 생약학회지
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    • 제53권3호
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    • pp.125-132
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    • 2022
  • In this study, we investigated anti-oxidative and anti-inflammatory efficacy, and identified their constituents from Brassica napus L. (Korean name: Yuchae) whole plant. Upon the anti-oxidative activities screening, the ethanol extract exhibited potent DPPH and ABTS+ radical scavenging activities. On the anti-inflammation studies using LPS-induced RAW264.7 cells, the extract inhibited the production of NO and pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) effectively. To identify major constituents of B. napus extract, further purification was performed and led to isolation of two compounds; isorhamnetin 3,7-O-diglucoside(1) and isorhamnetin 3-O-glucoside(2). Quantitative analysis by high pressure liquid chromatography (HPLC) determined the flavonoid 1 as the major constituent. Isolated compounds showed DPPH radical scavenging effects and decreased NO levels without causing cell toxicities. These results indicate that the extract of Yuchae, a rich plant resource in Jeju Island, could be potentially applicable as an anti-oxidative and/or anti-inflammatory ingredients.

미생물에 의한 $\beta$-Galactosidase의 생산 및 이용에 관한 연구 (제 1보) Penicillium sp.로부터 효소의 생산조건 및 정제 (Studies on the Production of $\beta$-Galactosidase by Microorganism and its Application (Part 1) Conditions for the Production and Purification of the Enzyme from Penicillium SP.)

  • 오평수;양한철
    • 한국미생물·생명공학회지
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    • 제9권4호
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    • pp.207-212
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    • 1981
  • 토양에서 분리된 여러가지 곰팡이류 가운데 $\beta$-galactosidase 분비력이 강하고 중성부근에서 효소안정성이 높은 Penicillium sp. 균주를 $\beta$-galactosidase 생산균주로 선정하였다. 이 균주는 밀기울고체배지에서 3$0^{\circ}C$ 72시간 배양에서 galactosidase 분비력이 가장 높았으며 질소원 첨가배지에서 질소원의 종류에 따라 14%~85%까지 증가되었다. 고체배지에서 얻은 효소추출액을 ammonium sulfate fractionation, SP-Sephadex C-50 chromatography, ultrogel AcA44 filtration, hrdroxrapatite chromatography등에 의하여 비활성도는 101 units/mg protein으로 5050배 정제되었다. 최종적으로 정제된 galactosidase는 analytical ultracentrifuge에서 단일단백으로 Schlieren pattern을 나타냈고 Disc electrophoresis에서는 단일 band로서 전기영동되었으며 영동된 $\beta$-galactosidase 활성 band와 일치하였다.

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Production of Lignin Peroxidase by Phellinus igniarius and Cytotoxic Effects of Lignin Hydrolysates Derived from Wood Biomass on Cancer Cells

  • Lee, Jae-Sung;Lee, Jong-Suk;Yoon, Jae-Don;Beak, Sung-Mok;Bosire, Kefa-O.;Lee, Yong-Soo;Kim, Jung-Ae
    • Biomolecules & Therapeutics
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    • 제12권3호
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    • pp.189-193
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    • 2004
  • Over the past several years, research efforts have been directed both at economically producing valuable substances from the wood biomass and at producing lignolytic enzymes at a lower cost. In the present study, we found that Phellinus igniarius, the basidiomycetes, secreted lignin peroxidase as a main lignolytic enzyme, which was detected maximum activity at 16th day of culture and showed 37 kDa of molecular mass in identification by activity assay and purification by anion-exchange chromatography. The Phellinus igniarius-derived lignin peroxidase hydrolyzed steam-exploded wood (Quercus mongolica) powder into small molecules showing cytotoxicity against cancer cel1s (HepG2 hepatoma, SK-N-SH neuroblastoma, B16 melanoma, MBT-2 bladder cancer). In addition, the enzyme hydrlysates of lignins (ELg) that were extracted from the steam-exploded oak showed more potent cytotoxic effects on the cancer cells than the enzyme hydrolysates of wood biomass (EWp), indicating that the cytotoxic effect of EWp may be due to the enzyme-degraded products of lignin among the lignocellulosics. Furthermore, the cytotoxic effect of ELg on Chang, normal liver cells, was much less potent than that of ELg on HepG2 and B16 cancer cells, indicating that the cytotoxic effect of ELg may be specific for cancer cells. The present results suggest that Phellinus igniarius may be a useful resource for the large-scale production of lignin peroxidase and that the lignin peroxidase may be applied for the generation of valuable biodegradation products from wood lignocellulosics for medical use.

대두유 응고효소 생산에 관한 연구 (Isolation and production of soymilk-clotting enzymes from Bacillus sp. K-324-7)

  • 이기성;한면수;심상국;정동효
    • Applied Biological Chemistry
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    • 제33권2호
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    • pp.154-160
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    • 1990
  • 토양 시료로 부터 선별과정을 거쳐 대두유를 응고시킬 수 있는 extracellular 대두유응고효소를 생산하는 균주를 분리하고 분리세균 K-324-7균주에 의한 효소생산의 확인과 형태적 생리적 성질을 조사한 결과 Bacillus cereus로 동정되었다. Bacillus K-324-7 분리균이 생산하는 대두유 응고효소는 황산암모늄 0.8포화 용액에서 가장 높은 활성을 나타내었으며, 최적활성 pH는 6.1이며, 최적활성온도는 $70^{\circ}C$이었다. 또한 pH 안정성은 pH $6.0{\sim}7.0$ 사이에서 안정하였고 열안정성은 $50^{\circ}C$ 이하의 온도에서 인정하였다. 한편 대두유 응고효소의 생산 배양조건을 검토한 결과 탄소원은 glucose 0.2%, 질소원은 peptone 0.2%, 무기염은 $KH_2PO_4$ 0.5%, pH는 6.5, 배양온도는 $35^{\circ}C$로 배양기간은 3일 정도가 적당하였으며 그 이상 배양하면 생성된 효소의 활성이 감소됨을 알 수 있었다.

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Purification and Characterization of an Exo-polygalacturonase from Botrytis cinerea

  • Lee, Tae-Ho;Kim, Byung-Young;Chung, Young-Ryun;Lee, Sang-Yeol;Lee, Chang-Won;Kim, Jae-Won
    • Journal of Microbiology
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    • 제35권2호
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    • pp.134-140
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    • 1997
  • Botrytis cinerea T91-1 has been shown to produce at least four different polygalacturonases into a liquid medium containing citrus pectin, a carbon sousrce. One of the enzymes, which had an apparent molecular weight of 66 kDa estimated by denatured polyacrylamide gel electrophoresis, was purified to electrophoretic homogeneity by a series of procedures including a cetone precipitation, ion exchange, heparin affinity, and reverse phase column chromatographies. The molecular weight of native enzyme was determined to be 64 kDa by gel permeation chromatography indicating the enzyme to be a single polypeptide chain. By viscometric analysis, the enzyme was revealed as exo-polygalacturonase. The enzyme activity was inhibited by divalent cations such as $Ca^{2+}$, $Mg^{2+}$, and Cu$^{2+}$. Km and Vmax for polygalacturonic acid hydrolysis were 0.33 mg/ml and 28.6 nM/min, respectively. The optimum temperature for enzymatic activity was 5$0^{\circ}C$. And the enzyme showed optimal pH values between 4.0 and 5.0. The enzyme was stable upto 12 hours in the range of pH 3 to 8 and at temperature below 3$0^{\circ}C$.

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고성능 폴리우레아 수지의 기본적 물리/화학적 성능평가에 관한 실험적 연구 (An experimental study on the Performance Evaluation of High Performance Polyurea Resins)

  • 김윤호;최은규;서현재;오상근
    • 한국건축시공학회:학술대회논문집
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    • 한국건축시공학회 2011년도 추계 학술논문 발표대회
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    • pp.43-45
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    • 2011
  • As technologies evolve, building large concrete structures ever built, but due to lack of maintenance after completion of concrete corrosion, leaks, and preparedness from the problem that is an urgent need. In particular, water-resistant variety of concrete structures. How the concept applies to the most important public drinking water purification and drinking water that is draining the production and storage, and distribution as the structure cause damage to the structure when the contaminated water is supplied to each home that can harm the health of citizens is the cause. Therefore, the correct choice of materials, and thorough a lot of investment in construction and maintenance should have. In this study, unlike conventional water-proof materials, methods, and in other reactions easily than conventional poly-urea resins have good physical performance and chemical resistance, high performance polyurea resin performance review of the physical infrastructure of the country for the longevity of would like to make long-term durability.

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A NEW TREATMENT SYSTEM FOR ANIMAL WASTE WATER USING MICROORGANISM, SOIL AND VEGETATION

  • Oshida, T.;Fukuyasu, T.;Kohzaki, K.;Izumikawa, Y.;Kawanabe, S.;Konishi, S.;Oikawa, N.;Matsumoto, S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제6권2호
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    • pp.205-209
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    • 1993
  • A new treatment system for animal waste water has been developed as an alternative to the activated sludge process. It consists of two treatments; one is operated with 7 tanks, and the other is soil and plant cultivation bed. Aerobic microorganisms are added to the influent water in the tanks where the water is aerated so that the microbes utilize the pollutants, while sedimentation removes the indigestible solids. In the secondary treatment the water, which has already received a primary treatment, is filtered through soil where it also receives treatment by soil organisms. In addition there is transpiration of water and absorption of minerals by plants. In the primary treatment BOD, SS, coliforms (E. coli), TP and total bacteria were removed 79-99%, but COD and TN were removed only 58% and 36%, respectively. In the secondary treatment removal of nutrients proceeded further, and 93-99% of pollutants were removed. The treated waters met the quality standard of discharge water in Japan except for TN, which was in too great a concentration to meet discharge standards. This problem requires further study.

인공갯벌 조성에 관한 경제적 타당성 분석 (Analyzing an Economic Feasibility for Restoration/Creation of Artificial Tidal Flats)

  • 남광현;오위영
    • Ocean and Polar Research
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    • 제25권4호
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    • pp.593-601
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    • 2003
  • The purpose of this paper is to investigate economic feasibility of creating artificial tidal flats using cost-benefit analyses. We assumed that the cost factors are associated with designing, construction and monitoring, and the benefit factors are associated with fisheries production, habitation, prevention of disasters, water purification, aesthetic value and existence value. First, for analyzing economic feasibility, the scenario suggests that a design can be made in a year, construction can be completed in three years and monitoring must be made for 20 years. Assuming the discount rate of 7.5%, economic feasibility analyses showed that B/C was 2.26 and IRR was 14.50. This study indicated there is economic validity of implementing creation of artificial tidal flat. In addition, we carried out a sensitivity analysis at the change of discount rate and restoration rate. The result of sensitivity analysis clearly showed that economic validity is low when discount rate is over 15%, and changes in restoration rate did not significantly effect on the economic validity.

Characterization of the Pediocin Operon of Pediococcus acidilactici K10 and Expression of His-Tagged Recombinant Pediocin PA-1 in Escherichia coli

  • MOON GI SEONG;PYUN YU RYANG;KIM WANG JUNE
    • Journal of Microbiology and Biotechnology
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    • 제15권2호
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    • pp.403-411
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    • 2005
  • The relationship between plasmid (~9.5 kb) and pediocin PA-1 in P. acidilactici K10 was confirmed by plasmid curing. The pediocin operon of P. acidilactici K10 was amplified by PCR (polymerase chain reaction), and the nucleotide sequence was analyzed. The sequence of the pediocin operon of P. acidilactici K10 was similar to those of P. acidilactici strains producing pediocin PA-1/ AcH. For the expression of pediocin PA-1 in E. coli, a pQEPED (pQE-30 Xa::mature pedA) was constructed. His-tagged recombinant pediocin PA-1 (-6.5 kDa) was translated by cell-free in vitro transcription and translation using pQEPED as a DNA template. Theresult of slot blotting assay showed that transcription of recombinant pedA in E. coli M15 was induced by the addition of isopropyl-$\beta$-D-thiogalactopyranoside (IPTG) at the final concentration of 1 mM. Although the recombinant pediocin PA-1 inhibited the growth of E. coli, it was expressed in the host strain and purified by nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography under denaturing condition. This is the first report for the production and one-step purification of biologically active recombinant pediocin PA-1 in E. coli.

재조합 균주 Escherichia coli가 생산하는 Bacillus stearothermophilus $\alpha$-L-Arabinofuranosidase의 정제 및 특성 (Purification and characterzation of the $\alpha$-L-Arabinofuranosidase from Escherichia coli Cells Harboring the Recombinant Plasmid pKMG11)

  • 엄수정;조쌍구;최용진
    • 한국미생물·생명공학회지
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    • 제23권4호
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    • pp.446-453
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    • 1995
  • $\alpha $-Arabinofuranosidase was produced by E. coli HB101 haboring the recombinant plasmid pKMG11 which contained the arfI gene of Bacillus stearothermophilus. The maximum production of the enzyme was observed when E. coli HB101 cells were grown at 37$\circ$C for 20 hours in the medium containing 0.5% arabinose, 1.0% tryptone, 0.5% yeast extract, and 1% NaCl. The $\ALPHA $-arabinofuranosidase produced was purified to homogeneity using a combination of 20-50% ammonium sulfate precipitation, DEAE-Sepharose CL-6B ion exchange column chromatography and Sepharose 6B-100 gel filtration. The purified enzyme was most active at 55$\circ$C and pH 6.5. The K$_{m}$ and V$_{max}$ values of the enzyme on $\rho $-nitrophenyl-$\alpha $-arabinofuranoside was determined to be 2.99 mM and 0.43 $\mu $mole/min (319.74 $\mu $mole/min/mg), respectively. The pI value was 4.5. The molecular weight of the native protein was estimated to be 289 kDa. The SDS-polyacrylamide gel clectrophoresis analysis suggested that the functional protein was a trimer of the 108 kDa identical subunits. The N-terminal amino acid sequence of the a-arabinofuranosidase was identified as X-Ser-Thr-Ala-Pro-Arg( \ulcorner )-Ala-Thr-Met-Val-Ile-Asp-X-Ala-Phe.

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