• 제목/요약/키워드: filteration

검색결과 91건 처리시간 0.021초

Molecular Cloning and Sequencing of the Bacillus subtilis cdd Gene Encoding Dooxycytindine-Cytidine Deaminase

  • Song, Bang-Ho;Neuhard, Jan
    • 한국미생물생명공학회:학술대회논문집
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    • 한국미생물생명공학회 1986년도 추계학술대회
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    • pp.512.1-512
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    • 1986
  • The cdd gene of Bacillus subtilis, encoding the deoxycytidinecytidine deaminase of pyrimidine nucleotide biosynthesis has been cloned into the EcoRl site of pBR322. The recombinant plasmid, pSol, promoted the synthesis of 100-140 fold elevated levels of the enzyme. A comparison of the polypeptides encoded by cdd complementing and non-complementing plasmids in the mini cell showed the gene product to have a molecular mass of approximately 14 kDa. The nucleotide sequence of the gene and 460 base pairs upstream from the coding region was determined. An open-reading frame, encoding a protein with a calculated molecular mass of 14337 Da, was deduced to be the coding region for cdd. However, the enzyme has an apparent molecular mass of 54 kDa as determined by gel filteration, whereas sucrose density gradient centrifugation shows 58 kDa. It means that the enzyme could be forming a tetramer in a physiological state. About 28 amino acids of the N-tetramer in a physiological state. About 28 amino acids of the N-terminal presumably form a signal for membrane translocation and six cystein residues are contained in the structure. S1 nuclease mapping indicated that transcription of cdd is initiated 17 base pairs upstream from the translational start. The structural characterization of the odd gene was performed.

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Streptomyces rimosus가 생산하는 Protease의 정제와 특성 (Purification and Properties of Extracellular Protease from Streptomyces rimosus)

  • 김경미;이태경;양한철
    • 한국미생물·생명공학회지
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    • 제17권5호
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    • pp.407-411
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    • 1989
  • Oxytetracycline 생산균주인 Streptomyces rimosus를 maltose 2%, NH$_4$Cl 0.5%, yeast extract 0.4 %, MGSO$_4$.7$H_2O$ 0.2% 조성의 배지를 배양초기 PH 6.5로 하여 3$0^{\circ}C$, 72시간 진탕배양하여 얻은, 세포외 protease를 유안분획, Sephadex A-50 이온교환, Sephadex G-100 gel 여과를 행하여 정제하였다. 효소의 최적온도는 5$0^{\circ}C$이며, 최적 pH는 8.0이었으며, Co$^{2+}$ 이온에 의해 활성화되며 Hg$^{2+}$, Fe$^{2+}$ 및 EDTA에 의해 저해를 받으며 casein 분자량을 23,600으로 추정하여 구한 Km값은 2.7$\times$$10^{-4}$M이었다.

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대두 단백 효소 가수분해물의 항균활성

  • 주정현;이상덕;이규희;이기택;오만진
    • 한국식품저장유통학회:학술대회논문집
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    • 한국식품저장유통학회 2003년도 춘계총회 및 제22차 학술발표회
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    • pp.145.1-145
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    • 2003
  • 대두 단백질을 효소로 가수분해 하였을 때 생성되는 peptide의 항균활성을 조사하고 천연항균제로서 이용 가능성을 검토하기 위하여 분리 대두 단백질에 5종의 단백질 가수분해 효소를 작용시켜 생성된 가수분해물의 항균력을 측정하고 한외여과하여 분자량별로 분리된 각 fraction의 항균활성과 HPLC로 정제하여 항균성 peptide 의 아미노산 결합순서를 분석하여 다음과 같은 결론을 얻었다. 분리대두 단백질에 5종의 단백질 분해효소를 작용시켜 제조한 가수분해물 중 Asp.saitoi protease로 작용시킨 것이 항균활성이 높았다. Asp. saitoi protease로 작용시킨 대두 단백질의 가수분해물을 membrane filter로 여과한 결과 분자량 1000-3000 fraction에서 항균활성이 가장 높았다. 분자량 1000-3000 범위을 가진 가수분해물의 MIC는 0.5-0.8mg/$m\ell$ 이었으며 그람 양성균과 음성균 모두의 증식을 억제하는 경향을 보였다. 분리 대두 단백으로부터 얻어진 항균성 peptide는 121$^{\circ}C$, 10분간 열처리하여도 안정하였으며 한외여과에 의하여 분자량 1000-3000범위의 가수분해물을 동결건조하여 gel filteration하였을 때 2개의 fraction에서 항균 활성을 나타내었다. HPLC결과 RT 16.02 의 peak에서 항균활성이 확인되었고 질량은 1,633이었으며 아미노산 결합순서는 H$_2$N-G-P-P-G-V-V-A-T-V-V-A-A-R-COOH 이었다.

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Cross-Flow Filtration에 의한 항생물질 생산 균사체의 분리 (Separation of Antibiotics-Producing Mycelia by Cross-Flow Filteration)

  • 정상철;윤종원
    • KSBB Journal
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    • 제4권2호
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    • pp.177-184
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    • 1989
  • 항생물질 발효액의 균사체 분리에 있어서 Modified regenerated cellulose 막을 이용한 Cross-flow filtration으로 높은 Flux를 얻을 수 있었다. 막분리시 가급적 높은 유속, 높은 온도, 특정 균사체농도(약 20% PMV)에서 높은 Flux를 보여주었으며 Fouling 정도는 균사체 농도가 높을수록 또한 유속이 낮을수록 심각한 것으로 나타났다. 발효액을 농축 분리할 때의 Flux profile은 1단계 Sublayer형성에 의한 급속감소, 2단계 평형, 3단계 Fouling에 의한 급속감소의 세 구간으로 나눌 수 있었으며 Diafiltration의 적용으로 3단계의 Flux 감소 현상을 현저히 완화시킬 수 있었다. 한편 Diafiltration에 의한 항생물질 회수시 초기부터Diafiltration하는 것보다는 특정 PMV까지 농축시킨 후 Diafiltration하는 경우에 작업시간과 여액의 Volume을 최소화 할 수 있었으며 본 실험에서는 98%의 회수율을 얻기 위해서 Di-afiltration을 시작하는 Optimum PMV는 20%였다.

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알긴산 분해 해양미생물의 분리 및 alginase 특성 평가 (Isolation of Alginate-Degrading Marine Bacteria and Characterization of Alginase)

  • 이재화;이은열
    • 생명과학회지
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    • 제13권5호
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    • pp.718-722
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    • 2003
  • 다양한 종류의 미역으로부터 알긴산 분해 활성을 가지는 해양 미생물인 Vibrio sp. AEBL-211을 분리하였다. 우선, 알긴산 고체배지에서의 halo 크기가 크고 환원당 생성량이 가장 많은 해양 미생물을 선발하고, 생화학 및 영양적인 특성 분석 결과 등을 바탕으로 Vibro 속으로 동정하였다. DE 52-cellulose 및 Sephacryl G-200를 이용한 음이온 교환 및 gel permeation chromatography를 통해 부분 정제된 alginase 효소액을 얻었다. 부분 정제된 alginase 효소를 이용하여 guluronic acid에 대한 mannuronic acid의 비율이 다른 sodium alginate 시료에 대한 분해능을 평가한 결과, guluronic acid의 상대적 양이 많은 알긴산에 대한 활성이 상대적으로 우수하여 Vibrio sp. AEBL-211 유래의 alginase는 guluronic acid가 많은 G-rich block에 대한 분해활성이 높은 특성을 가지는 것으로 판단되었다.

안쓰라사이트 여재 원형도가 여과 효율에 미치는 영향 (The Effects of Anthracite Media Sphericity on Filtration Efficiency)

  • 정원석;최승일
    • 상하수도학회지
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    • 제21권6호
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    • pp.763-770
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    • 2007
  • There are many design parameters affecting filtration efficiency such as filteration rate, media packing depth, size distribution, and so on. The sphericity, the ratio of the surface area of an equal volume sphere to the real surface area of the particles, is one of major physical characters of media. The effect of sphericity on the performance of anthracite filter has been investigated. Media from eight water treatment plants have been collected. The sphericity of each media has been calculated by using well known headloss equations such as Kozeny equation, Dahmarajah equation etc.. Columns packed with anthracite media having different sphericity have been used to compare headloss development, floc accumulation in the bed, particles in bed water, filtrate turbidities after backwash and so on. The repeated experiments have indicated that the sphericity of anthracite media may not have remarkable influence on the filter performance as it has been suspected. It also has been prospected in the experiment that the media of higher sphericity would store more particles in the bed and give better filtrate quality, if provided that the effective size and the size distribution of media would be the same.

에멀젼을 이용한 $Al_2O_3$-$ZrO_2$복합분체의 제조 : III. 에멀젼-공침법 (Preparation of $Al_2O_3$-$ZrO_2$ Composite Powders by the Use of Emulsions : III. Emulsion-Coprecipitation Method)

  • 현상훈;김의수
    • 한국세라믹학회지
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    • 제27권4호
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    • pp.487-494
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    • 1990
  • $Al_2O_3$-20w/o $ZrO_2$ composite powders were prepared by the emulsion-coprecipitation method and the effects of preparative conditions on powder characteristics were investigated. In the preparation of $Al_2O_3$-$ZrO_2$ composite powders, toluene was used instead of kerosene as the oil phase in emulsions. After coprecipitation, the emulsion was easily broken into a single liquid phase by adding methanol, and then precipitates could be effectively collected by filteration. The fact that all $ZrO_2$ phases present at room temperature in composite powders calcined at $1100^{\circ}C$ after washed by methanol had a tetragonal structure confirmed that methanol-washing enhanced the dispersibility of fine $ZrO_2$ particles in $Al_2O_3$ matrix. $Al_2O_3$-$ZrO_2$ composite powders were spherical particles of 0.2${\mu}{\textrm}{m}$ diameter. Pellets sintered at $1650^{\circ}C$ for 2hrs showed the relative theoretical density of 97.3% and the fracture toughness of 5.01MN/$m^{3/2}$.

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One-step purification and biochemical characterization of a (s)-stereospecific esterase from Pseudomonas fluorescens KCTC 1767

  • 최기섭;김지희;김지연;김근중;유연우
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2002년도 생물공학의 동향 (X)
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    • pp.445-448
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    • 2002
  • The Pseudomonas fluorescens KCTC 1767, a selected and identified as potential candidate for stereo-specific resolution of rac-ketoprofen ethyl ester, was systematically investigated in order to induce the high level expression and detailed characterization of the expressing enzyme esterase. We cloned the esterase gene from chromosomal DNA of Pseudomonas fluorescens KCTC 1767 by PCR with two synthetic primers that desinged for simple purification. The recombinant esterase from Pseudomonas fluorescens KCTC 1767 exibited a high conversion rate and enantioselectivity to the (S)-ketoprofen ethyl ester as expected. The enzyme was easily purified to homogeniety by using a metal chelating affinity chromatography as a protein with poly histidine taq, and thus obtained 0.6 mg of protein from a 100 mL culture broth in a single step. The purified enzyme was steadily stable at the pH range from 7.0 to 10. The activity was also retained to be about 70% after the preincubation at $40^{\circ}C$ but over $50^{\circ}C$ lost the activity completely. The molecular mass of the esterase was estimated to be about 43 kDa on SDS-PAGE, and an identical result was also shown in gel filteration chromatography. The specific activity was calculated 27 mM/mg-protein/min by using the rac-ketoprofen ethly ester as a substrate.

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간이상수도 자동소독 정수기의 개발 및 성능에 관한 연구 (Development and Performance of Water Purifier with the Auto-Disinfected on a simple Drinking Water)

  • 조병락;이배복;최명부
    • 한국산업융합학회 논문집
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    • 제16권2호
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    • pp.41-46
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    • 2013
  • On the purpose of helping the inhabitants living in farming, fishing villages, and islands for more safe and hygienic water from simple waterworks, experimental investigations were performed concerning the development of a water purifier with silver nanomaterial packed, having a function of the auto-disinfection. The results show as follows through such filteration and auto-disinfection processes. It is possible to get hygienic and safe water, for example, more than 95% of general bacteria, total coliforms, and fecal coliforms were removed. It is also possible to get good-quality water, for 49.4% of spent potassium permanganate and 85% and 63% of turbidity and conductivity were removed respectively. It is a very effective equipment, for 100% cost reduction of used chemicals was achieved by no-chemical disinfection process and THM was not generated.

뱀 근육(筋肉) Adenosine Triphosphate - Creatine Phosphotransferase에 관(關)한 연구(硏究) (Studies on Adenosine Triphosphate - Creatine Phosphotransferase from Muscle of the Snake Bungarus fasciatus)

  • 박충웅
    • 한국식품영양과학회지
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    • 제9권1호
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    • pp.59-73
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    • 1980
  • A detailed procedure was described for the isolation of cratine kinase (ATP-Creatine phosphotransferase, E. C. 2. 7. 3. 2.) from the muscle of the snake Bungarus fasciatus. The original isolation procedure of Kuby et al. for the rabbit muscle enzyme has been modified and extended to include a chromatographic step. The properties of the enzyme have been investigated and kinetic constants for the reverse reactions determined as the followings: 1) A molecular weight of the enzyme was determined by gel filteration on Sephadex G-100 and by electrophoresis on SDS-polyacrylamide was 86,000. 2) Two reactive sulphydryl groups were detected with dithiobis nitrobenzoic acid (DTNB). 3) The nucleotide substrate specificity in the reverse reaction was determined as ADP*2'-dADP>GDP>XDP>UDP with magnesium as the activating metal ion. 4) The order of the metal specificity in the reverse reaction Mg>Mn>$Ca{\sim}Co$ was determined with ADP as substrate. 5) A detailed kinetic analysis was carried out in the reverse direction with $MaADP^-$ as the nucleotide substrate. Initial velocity and product inhibition studies($MaADP^{2-}$ competitive with respect to MgADP- and noncompetitive with respect to $N-phosphorycreatine^{2-}$ ; Creatine competitive with respect to $N-phosphorycreatine^{2-}$ and noncompetitive with respect to Ma $ADP^-)$ indicated that the reaction obeyed a sequential mechanism of the rapid equilibrium random type.

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