• 제목/요약/키워드: protein removal

검색결과 324건 처리시간 0.032초

Performance of foam fractionator in seawater recirculating system

  • Lei Peng;Jo, Jae-yoon
    • 한국어업기술학회:학술대회논문집
    • /
    • 한국어업기술학회 2003년도 춘계 수산관련학회 공동학술대회발표요지집
    • /
    • pp.221-222
    • /
    • 2003
  • Typically, closed production system units are subject to an accumulation of fine suspended solids and dissolved organics (Weeks et at., 1992). Foam fractionation process is believed to be most effective in marine application for solids removal. In present experiment, the performance of foam fractionator for removal of solids, protein, and other dissolved materials was evaluated at different foam overflow heights and air flow rates in a pilot-scale recirculating aquaculture system for culture of Korean rockfish. (omitted)

  • PDF

Novel Modification of Growth Medium Enables Efficient E. coli Expression and Simple Purification of an Endotoxin-Free Recombinant Murine Hsp70 Protein

  • Zachova, Katerinat;Krupka, Michal;Chamrad, Ivo;Belakova, Jana;Horynova, Milada;Weigl, Evzen;Sebela, Marek;Raska, Milan
    • Journal of Microbiology and Biotechnology
    • /
    • 제19권7호
    • /
    • pp.727-733
    • /
    • 2009
  • Heat shock protein 70 kDa (hsp70), a molecular chaperone involved in folding of nascent proteins, has been studied for its ability to activate innate and specific immunity. High purity hsp70 preparation is generally required for immunization experiments, because endotoxins and other immunologically active contaminants may affect immune responses independently of hsp70. We have developed a novel modification of E. coli-expression medium that enabled a simple two-step production and purification method for endotoxin-free recombinant hsp70. During Ni-NTA-based affinity purification of hsp70, a contaminating protein from host E. coli cells, L-glutamine-n-fructose-6-phosphate aminotransferase (GFAT), was identified. By testing various compounds, supplementation of growth medium with a GFAT metabolite,N-acetylglucosamine, was found to reduce GFAT expression and increase the total hsp70 yield five times. The new protocol is based on column purification of His-tagged hsp70 protein produced by E. coli with the modified medium, followed by endotoxin removal by Triton X-114 extraction. This approach yielded hsp70 with high purity and minimal endotoxin contamination, making the final product acceptable for immunization experiments. In summary, a simple modification of growth medium allowed production of recombinant mouse hsp70 in high yield and purity, thus compatible with immunological studies. This protocol may be useful for production of other Histagged proteins expressed in E. coli.

Analysis of a Large-scale Protein Structural Interactome: Ageing Protein structures and the most important protein domain

  • Bolser, Dan;Dafas, Panos;Harrington, Richard;Schroeder, Michael;Park, Jong
    • 한국생물정보학회:학술대회논문집
    • /
    • 한국생물정보시스템생물학회 2003년도 제2차 연례학술대회 발표논문집
    • /
    • pp.26-51
    • /
    • 2003
  • Large scale protein interaction maps provide a new, global perspective with which to analyse protein function. PSIMAP, the Protein Structural Interactome Map, is a database of all the structurally observed interactions between superfamilies of protein domains with known three-dimensional structure in thePDB. PSIMAP incorporates both functional and evolutionary information into a single network. It makes it possible to age protein domains in terms of taxonomic diversity, interaction and function. One consequence of it is to predict the most important protein domain structure in evolution. We present a global analysis of PSIMAP using several distinct network measures relating to centrality, interactivity, fault-tolerance, and taxonomic diversity. We found the following results: ${\bullet}$ Centrality: we show that the center and barycenter of PSIMAP do not coincide, and that the superfamilies forming the barycenter relate to very general functions, while those constituting the center relate to enzymatic activity. ${\bullet}$ Interactivity: we identify the P-loop and immunoglobulin superfamilies as the most highly interactive. We successfully use connectivity and cluster index, which characterise the connectivity of a superfamily's neighbourhood, to discover superfamilies of complex I and II. This is particularly significant as the structure of complex I is not yet solved. ${\bullet}$ Taxonomic diversity: we found that highly interactive superfamilies are in general taxonomically very diverse and are thus amongst the oldest. This led to the prediction of the oldest and most important protein domain in evolution of lift. ${\bullet}$ Fault-tolerance: we found that the network is very robust as for the majority of superfamilies removal from the network will not break up the network. Overall, we can single out the P-loop containing nucleotide triphosphate hydrolases superfamily as it is the most highly connected and has the highest taxonomic diversity. In addition, this superfamily has the highest interaction rank, is the barycenter of the network (it has the shortest average path to every other superfamily in the network), and is an articulation vertex, whose removal will disconnect the network. More generally, we conclude that the graph-theoretic and taxonomic analysis of PSIMAP is an important step towards the understanding of protein function and could be an important tool for tracing the evolution of life at the molecular level.

  • PDF

광생물반응기내 광조사 유형별 탈황 효율 (Desulfurization efficiency in photobioreactors dependent on the irradiation type of light sources)

  • 안진영;김병우
    • 청정기술
    • /
    • 제6권1호
    • /
    • pp.71-78
    • /
    • 2000
  • 본 연구는 광합성 균주인 C. thiosulfatophium을 이용한 황화수소의 생물학적 탈황공정시 광에너지를 저감시키기 위해 광원의 종류와 광조사 유형에 따른 탈황효율을 조사하였다. 광에너지를 저감시키기 위해서는 반응기내 광분산 효율을 극대화시켜야 한다. 광이용효율을 높이기 위해, 세균농도와 생성부산물의 증가에 의한 빛의 산란과 흡수가 세균성장에 미치는 영향에 관한 연구와 미생물이 필요로 하는 파장의 빛을 최대로 공급하기 위해 LED, 백열등, 태양광 등을 이용한 광원의 최적화 연구, 그리고 외부조사형 광반응기의 광투과 효율의 한계를 극복하기 위한 광섬유를 이용한 내부조사형 광반응기에 대한 연구를 수행하였다.

  • PDF

Quality Control of Photosystem II during Photoinhibition

  • Yamamoto, Yasusi
    • Journal of Photoscience
    • /
    • 제9권2호
    • /
    • pp.55-58
    • /
    • 2002
  • The reaction center Dl protein of photosystem II is the target of photodamage by excess illumination. The Dl protein is damaged by reactive oxygen species generated by photochemical reactions and then degraded by specific proteolytic enzymes. We found that the Dl protein also cross-links with the surrounding polypeptides, such as D2 and CP43 in isolated thylakoids or photosystem II-enriched membranes from spinach under the illumination with strong visible light. The cross-linking was observed in spinach leaf discs as well when they were illuminated at higher temperature (40°C). It was also shown that the cross-linked products are digested efficiently by a protease(s) in the stroma. Thus the cross-linking/digestion processes of the Dl protein seem to comprise a new pathway in the turnover of the photodamaged Dl protein. It should be noted, however, that the cross-linked products of the Dl protein and CP43 induced by endogenous cationic radicals in the donor-side photoinhibition are resistant to proteolytic digestion. Accumulation of these cross-linked products in the thylakoids may lead to the decay of the function of chloroplasts and finally to the death of plant cells. Thus, we suggest that the quality control of photosystem II, especially removal of the cross-linked products of the Dl protein, is crucial for the survival of chloroplasts under the light stress.

  • PDF

적엽ㆍ제협처리가 콩의 광합성과 동화물질 배분에 미치는 영향 (Effects of Leaf and Pod Removal on Photosynthesis and Assimilate Partition in Soybean)

  • 김웅태;성락춘;해리 씨 마이너
    • 한국작물학회지
    • /
    • 제38권2호
    • /
    • pp.159-165
    • /
    • 1993
  • 대두에서 각각 50%의 협제거와 협제거 처리에 의해서 변화된 동화물질에 대한 sink demand가 엽의 광합성율과 각 부위별 단백질 함량 및 엽의 노화에 미치는 영향을 구명하기 위하여, 1990년 고려대학교 자연자원대학, 식량자원학과 실험포장에서, pot에 팔달콩을 공 시하여 총건물중과 엽을 제외한 vegetative tissues의 건물중, 잎과 뿌리의 전분함량, 엽의 광합성율과 기공저항성, 각 부위별 단백질함량, 엽의 chlorophyll content등을 조사한 결과를 요약하면 다음과 같다. 1. 엽을 제외한 vegetative tissues의 건물중은 협제거 처리에 의해서 증가되었고, 따라서 협제거 처리에 의한 총건물중의 감소는 유의성이 없었다. 2. 종실의 100립종은 대조구와 비교할 때, 50% 협제거 처리에 의해 6.4% 증가하였으며, 50% 엽제거 처리에 의해서 3.0% 감소하였다. 3. 엽의 전분함량은 처리 후 22일부터 감소되었고 뿌리의 전분함량은 전 시기에 걸쳐 감소되었는데, 잎과 뿌리의 전분함량은 협제거 처리에 의해 높아지고 엽제거 처리에 의해 낮아지는 경향을 보였다. 4. Specific leaf weight는 협제거 처리에 의해 증가되었고 엽제거에 의해 감소 되었다. 5. 엽의 광합성율은 전 시기에 걸쳐서 높아졌으며, 협제거 처리에 의해 증가되었다. 7. 잎, 중기+엽병, 뿌리 등의 단백질함량은 점차로 감소되는 경향이었으며, 협제거 처리에 의해 높아졌고 엽제거에 의해 낮아지는 경향을 보였다. 8. 성숙기( $R_{7}$)의 종실의 단백질함량은 협제거와 엽제거 처리에 의해 유의성 있는 차이를 나타내지 않았다. 9. 엽의 chlorophyll content의 감소에 의한 엽의 외관상 노화는 협제거처리에 의해 지연되었고 엽제거에 의해 촉진되었다. 10. 광합성율의 감소에 의한 엽의 기능상 노화는 엽제거 처리에 의해 촉진되었다.

  • PDF

Precise assembly and regulation of 26S proteasome and correlation between proteasome dysfunction and neurodegenerative diseases

  • Im, Eunju;Chung, Kwang Chul
    • BMB Reports
    • /
    • 제49권9호
    • /
    • pp.459-473
    • /
    • 2016
  • Neurodegenerative diseases (NDs) often involve the formation of abnormal and toxic protein aggregates, which are thought to be the primary factor in ND occurrence and progression. Aged neurons exhibit marked increases in aggregated protein levels, which can lead to increased cell death in specific brain regions. As no specific drugs/therapies for treating the symptoms or/and progression of NDs are available, obtaining a complete understanding of the mechanism underlying the formation of protein aggregates is needed for designing a novel and efficient removal strategy. Intracellular proteolysis generally involves either the lysosomal or ubiquitin-proteasome system. In this review, we focus on the structure and assembly of the proteasome, proteasome-mediated protein degradation, and the multiple dynamic regulatory mechanisms governing proteasome activity. We also discuss the plausibility of the correlation between changes in proteasome activity and the occurrence of NDs.

포말 농축물에 의한 포말 생성의 영향인자 (The Effective Factors of n Foam Generation Using Foam Condensate)

  • 서근학;신정식;이주화
    • 한국수산과학회지
    • /
    • 제36권5호
    • /
    • pp.509-514
    • /
    • 2003
  • We performed the experiment to determine the effective factors, such as the initial concentration of protein, pore size of air distributor, SAV (superficial air velocity), pH, salts and temperature related to foaming characteristics. The foam height in a foam generator was increased with the increase of the initial protein concentration and the decrease of pore size. As SAV was increased, the foam height was increased, and the optimum SAV was 0.84 cm/sec. The foam height was highest in the acid region and it was increased with the increase of salt concentration of NaCl and $NaHCO_3.$ The removal efficiencies of TSS (total suspended solid) and turbidity decreased with the increase of the initial protein concentration in the batch foam separator.

Preparation of Blood Glue from Porcine Plasma Protein and Cross-linking Reaction of Plasma Protein with Formaldehyde

  • Cho, Yongsik;Lee, Hwahyoung;Song, Kyung Bin
    • Journal of Applied Biological Chemistry
    • /
    • 제42권2호
    • /
    • pp.81-84
    • /
    • 1999
  • Blood glue was prepared to reutilize porcine blood. Plasma proteins after lyophilization were treated by addition of wood flour, sodium hydroxide, sodium silicate, and hydrated lime to make blood glue with a suitable adhesivity. Characteristics of the prepared blood glue was monitored by measuring the viscosity with time, and the relationship between degree of hydrolysis of plasma proteins by addition of various amounts of sodium hydroxide and adhesivity was studied. To prevent the emission of formaldehyde during manufacturing of plywood by blood glue, the cross-linking reaction of plasma protein with formaldehyde was also examined. Fourier transform infrared, circular dichroism, and fluorescence spectroscopy study showed that blood plasma proteins react with formaldehyde, resulting in removal of formaldehyde by cross-linking reaction.

  • PDF

Single-Step Purification of Proteins of Interest from Proteolytically Cleaved Recombinant Maltose-binding Protein (MBP) Fusion Proteins by Selective Immunoprecipitation of MBP

  • Park, Jung-Hyun;Na, Shin-Young;Lee, Dong-Gun;Han, Byoung-Don;Kim, Kil-Lyong
    • Biotechnology and Bioprocess Engineering:BBE
    • /
    • 제3권2호
    • /
    • pp.82-86
    • /
    • 1998
  • The maltose binding protein (MBP) fusion protein system is a versatile tool to express and isolate recombinant proteins in E. coli. In this system, MBP fusion proteins are efficiently isolated from whole cell lysate using amylose conjugated agarose beads and then eluted by competition with free maltose. Since MBP is a rather large molecule (∼42 kDa), for further experiments, the MBP part is usually proteolytically cleaved from the fusion protein and subsequently removed by ion-exchange chromatography or rebinding to amylose columns after washing out excess and MBP-bound maltose. In the present study, we have developed an improved method for the removal of cleaved MBP, which is advantageous over conventional methods. In this method, factor Xa cleaved MBP fusion proteins were incubated with Sepharose beads conjugated with MBP specific monoclonal antibodies and then precipitated buy centrifugation, resulting in highly purified proteins in the supernatant.

  • PDF