• Title/Summary/Keyword: 겔 전기영동

Search Result 77, Processing Time 0.029 seconds

돈피 콜라겐 유래 올리고펩타이드 제조를 위한 방사선조사의 이용

  • Jo, Yeong-Jun;Seo, Jeong-Eun;Kim, Yeong-Ho;Kim, Yun-Ji;Lee, Nam-Hyeok;Hong, Sang-Pil
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
    • /
    • 2005.10a
    • /
    • pp.150-152
    • /
    • 2005
  • 환경친화적 기술로 알려진 방사선조사기술(RT: Radiation Technology)을 이용하여 돈피 유래 올리고펩타이드를 제조하고자 하였다. 생 박 돈피를 hammer mill과 chopper를 이용하여 조분쇄한 후 $-20^{\circ}C$ 아세톤으로 탈지하였고, ${\gamma}$-ray irradiator를 이용하여 0, 20, 40, 60, 100, 150, 200, 250, 300 kGy의 총 흡수량을 얻도록 탈지돈피에 방사선조사를 실시하였다. 방사선조사에 의한 탈지돈피의 pH 변화는 0${\sim}$100 kGy 조사선량에서는 미비했으나, 150 kGy 이상에서는 소폭 증가하였다. 탈지돈피의 단백질 함량 중 콜라겐 함량은 93% 이었으며 방사선조사된 돈피콜라겐을 효소처리하면 효소반응 시간이 길어질수록 약 24 kDa 범위에서 밴드가 확인되었고, 100 kGy 이상의 고선량에서는 효소반응 2시간 이후 10% polyacrylamide 전기영동 겔의 최 하단에 머무는 분자량의 펩타이드가 다량 관찰되었다. 용해도 변화는 20${\sim}$60 kGy의 선량에서는 효소반응 시간이 길어질수록(1시간${\sim}$4시간) 최대 65${\sim}$80%의 용해도 증가를 보였고, 반면에 100 kGy 이상에서는 효소반응 시간에 관계없이 80% 이상, 300 kGy에서는 90% 이상의 용해도를 보여주려다. 점도와 탁도는 100 kGy 이상의 고선량에서 짧은 효소반응 시간(1시간)에 급격히 감소하였다. 가수 분해물(300 kGy)을 gel permeation chromatography한 결과 분자량 9,000 Da의 주 피크가 검출되었다.

  • PDF

Identification of Lactic Acid Bacteria from Meat by Low Molecular Weight(LMW) RNA Profiles (Low Molecular Weight(LMW) RNA Profiles에 의한 젖산균의 동정)

  • Cha, Woen-Suep
    • Journal of the Korean Society of Food Science and Nutrition
    • /
    • v.21 no.6
    • /
    • pp.681-685
    • /
    • 1992
  • Low molecular weight RNA(LMW RNA : 5S rRNA and tRNAs, <150 nucleotides) profiles of several bacteriocin production lactic acid bacteria from pig meats and reference lactic acid bacteria were generated on 10% denaturing polyacrylamide gel electrophoresis. Data evaluation including three molecular weight markers enabled the calculation of relative nucleotide units(RNU) for every band. Gels profiles and RNU evaluations were effective for identification of lactic acid bacteria species. LMW RNA profiles of lactic acid bacteria showed no variation in dependence on APT(All Purpose Tryptone Broth), TSB(Tryptic Soy Broth), MRS(Lactobacilli MRS Broth) different cultural medium.

  • PDF

Comparative Electrophoretic Studies of Isoperoxidase for 23 Species in the Genus Pinus (소나무속(屬) 23수종(樹種)에 있어서 Peroxidase 동위효소(同位酵素)의 변이(変異))

  • Park, Young Goo
    • Journal of Korean Society of Forest Science
    • /
    • v.51 no.1
    • /
    • pp.36-40
    • /
    • 1981
  • The patterns of isoperoxidase in needle-leaves of 23 species of the genus Pinus were analyzed by means of starch gel electrophoresis. Each species had a unique band pattern. In all, 56 isoperoxidase bands were identified, of which 9 to 35 isoperoxidase bands were possessed by single species. No single band was common to all Pinus species but when band patterns were grouped into 7 types, type II was considered to be the specific to genus Pinus. The results of this experiment indicated that various Pinus species had their more or less specific band patterns of peroxidase.

  • PDF

Purufication and Characterization of Extracellular Collagenase from Vibrio mimicus (Vibrio mimicus 가 생산하는 collagenase의 정제 및 특성)

  • 김용태;김세권
    • Journal of Life Science
    • /
    • v.6 no.4
    • /
    • pp.241-249
    • /
    • 1996
  • A collagenase was isolated from the culture filtrate of Vibrio mimicus (ATCC 33658). The enzyme was purified to homogeneity by ammonium sulfate precipitation and DEAE-Sephadex A-50 chromatography, which an activity recovery of 22%. The molecular weight of the purified enzyme was estimated to be 42 kDa by SDS-polyacrylamide gel electrophoresis and gel filtration, indication a monomer structure. The optimum pH and temperature od the enzyme for insoluble collagen (Type I) were around 7.75 and 28$\circ$C, respectively. Some chelating agents and serine protease inhibitor inactivated the enzyme, but L-cysteine and histidine did not affect the activity. The amino acid composition indicated that the collagenase contained high amounts of amino acid residues of glycine and alanine. The K$_{m}$ and R$_{cat}$/K$_{m}$ values for the collagenase, using insoluble collagen (type I) as substrate, were 2.86 mg/ml and 972.28 U/mg-protein, respectively.

  • PDF

Separation of Highly Purified Antimicrobial Lysozyme Using Ultrafiltration and Characteristics of Membrane Fouling (한외여과 공정을 이용한 고순도 향균 Lysozyme 의 분리 및 막 침착 특성)

  • Lee, Eun-Young;Woo, Gun-Jo
    • Korean Journal of Food Science and Technology
    • /
    • v.31 no.2
    • /
    • pp.458-464
    • /
    • 1999
  • The value of lysozyme as a natural food preservative is continuously increased due to its unique antimicrobial activity. To determine the optimum separation concentration among the various hen egg white protein (HEWP) concentrations (0.25, 0.5, 1.0, w/v), protein concentrations, lysozyme concentrations, specific activities (SA), and purification factors of prefiltered solution (PFS) and PM30 permeate solution (PMS) were compared. The purity of lysozyme separated at each step was analyzed and confirmed by gel permeation chromatography and electrophoresis. The fouling deposits on membrane were observed by SEM. The non-enzymatic proteins were removed over 99% by ultrafiltration (UF). The increased feed concentration did not contribute to the increase of SA. SA of PMS was 18 to 31 times higher than that of PFS. The optimum feed concentration was decided as 0.25% based on SA and purification factor. The non-enzymatic region of gel chromatogram was proved to be ovalbumin. The thickness of deposit on the UF membrane was approximately $0.9{\mu}m$ and removed by cleaning with 0.1 N NaOH. Therefore, UF using PM30 membrane was very effective to separate the antimicrobial lysozyme from various HEWPs.

  • PDF

Identification and characterization of allergens of Chironomus fkavuoynys adults (Chironomidae, Diptera) in mice (노랑털깔따구(Chironomus flaviplumus) 성충의 알레르기 항원단백 분석)

  • 이한일;이상화
    • Parasites, Hosts and Diseases
    • /
    • v.34 no.1
    • /
    • pp.35-48
    • /
    • 1996
  • Non-biting midges fchironomidae, Dipteral are one of the largest insect families, which are distributed worldwidely and are found in nearly all types of inland waters. They are known to be aggressive inhalant allergens which cause allergenic diseases. In this study, the crude antigens of Chironomus SavipLumn adults which are most widely distributed in Korea were extracted. and their allergens were analysed with the sera from experimentally sensitized mice. The mice were immunized with $1{\;}\mu\textrm{g}{\;}or{\;}10{\;}\mu\textrm{g}$ of the crude antigens, respectively, and the specific serum IgE levels were measured by both ELISA and passive cutaneous anaphylaxis (PCA) techniques. The highest levels of both total IgE and chironomid-specific IgE were found in the mouse sera obtained after 9 weeks of the first infection with $1{\;}\mu\textrm{g}$ crude antigen. The crude antigen was separated into 16-18 protein bands on gel by SDS-PAGE. The crude extract was assessed by SDS-PAGE/immunoblot analysis. One IgE-binding band (65 kDa) was detected by developing with colorimetric substrate, and 4 IgE-binding bands (65, 52, 35 and 25 kDa) by developing with CSPD chemiluminescent substrate. The SDS-PAGE gel of the crude extract of chironomid adults was equally cut into 30 pieces and each of them was eluted to isolate proteins by molecular weight, and the allergenicity of each eluate was assessed by applying P-K test on rats. Proteins of 65, 35 and 15 KDa showed the highest P-K titer (${\times}512$) which was 16 times higher than that of the crude extract (${\times32}$). The P-K titer of 52 kDa protein was also 4 times higher ($128{\times}$) than that of the crude extract, whereas the 25 kDa protein poorly responded, which seemed not antigenic. In conclusion, the present result in mice demonstrated that adults of Chironomus fcuiplumus, a predominent species in Korea, cause allergenic diseases and the main allergens are 65, 52, 35 and 15 kDa proteins, of which 65 kDa protein seems to be a main allergen.

  • PDF

Utilization of Pigments and Tunic Components of Ascidian as an Improved Feed Aids for Aquaculture 2. Chemical Properties of Sulfated Polysaccharides in Ascidian (Halocynthia roretzi) Tunic (우렁쉥이 껍질성분 및 색소를 이용한 양식소재 개발 2. 우렁쉥이 껍질 중 부분정제한 황산다당의 화학적 특성)

  • HONG Byeong-Il;JUNG Byung-Chun;JUNG Woo-Jin;RUCK Ji-Hee;CHOI Byeong-Dae;LEE Kang-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.34 no.6
    • /
    • pp.632-637
    • /
    • 2001
  • Components of polysaccharides isolated from ascidian tunic were measuerd by gel filtration, electrophoresis and chemical analyses. The sulfated polysaccharides consisted in sulfate, protein, uronic acid and amino sugars. Hexosamines were composed of arabinose, xylose, glucose, galactose, glucuronic acid, N-acetylgalactosamine and N-acetylglucosamine by gas chromatography analysis. The galactose was predominant hexose after autoclave and nutrase digestion followed by DEAE-cellulose ion exchange chromatography and gel-permeation chromatography on Sephadex G-100 and G-25. FT-IR spectra of isolated polysaccharides from ascidian tunic and standard chondroitin sulfates have similar functional groups of the type of vibration and frequency. Molecular weight of isolated polysaccharides by autoclave represented more than 40 kDa by polyacrylamide gel electrophoresis. But the neutrase treatment group divided into three band. The highest molecular band group was shown more than 100 kDa, and the two low molecular band group were shown about 22 kDa and 5 kDa, respectively, compare to standard materials.

  • PDF

Rheological Properties of Pork Myofibrillar Protein and Sodium Caseinate Mixture as Affected by Transglutaminase with Various Incubation Temperatures and Times (Transglutaminase를 첨가한 돈육 근원섬유단백질과 카제인염 혼합물의 배양온도와 시간에 따른 물성변화)

  • Hwang, Ji-Suk;Lee, Hong-Chul;Chin, Koo-Bok
    • Food Science of Animal Resources
    • /
    • v.28 no.2
    • /
    • pp.154-159
    • /
    • 2008
  • To investigate the rheological properties of protein mixed gels mediated by microbial transglutaminase (MTGase), pork myofibrillar protein (MFP), sodium caseinate (SC) and their mixture (MS), the various gels were incubated at different temperatures for various times. Extracted MFP, SC and their mixture (MS, 1:1) were incubated at different temperatures ($4^{\circ}C$ vs $37^{\circ}C$) for various times (0, 0.5, 2, 4 hr), and assessed for viscosity, gel strength and other characteristics using differential scanning calorimeter (DSC) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). DSC measurements showed that incubation at $37^{\circ}C$ rather than $4^{\circ}C$ caused marked changes in thermal transition, and MS displayed similar thermal curves (three endothermic transitions) to MFP and SC alone. After incubation at $37^{\circ}C$ for 2 hrs, the viscosity (cP) of MS increased (p<0.05) due to induction by MTGase, whereas no differences were observed at $4^{\circ}C$. However, gel strength values were no different, regardless of incubation temperatures and times. Future research will address how longer incubation times affect the functionality of protein mixed gels mediated by MTGase.

Study on Microbiochip for Buccal Cell Lysis and DNA Purification (상피세포 시료 전처리용 마이크로바이오칩에 관한 연구)

  • Ha, Seung-Mo;Cho, Woong;Ahn, Yoo-Min;Hwang, Seung-Yong
    • Transactions of the Korean Society of Mechanical Engineers A
    • /
    • v.34 no.12
    • /
    • pp.1785-1791
    • /
    • 2010
  • This paper describes a separable microfluidic device fabricated with PDMS (polydimethylsiloxane) and glass. The device is used for sample preparation involving cell lysis and the DNA purification process. The cell lysis was performed for 2 min at $80^{\circ}C$ in a serpentine-type microreactor ($20 {\mu}l$) using a Au microheater that was integrated with a thermal microsensor on a glass substrate. The DNA that was mixed with other residual products during the cell lysis process was then filtered through a new filtration system composed of microbeads (diameter: $50 {\mu}m$) and PDMS pillars. Since the entire process (sample loading, cell lysis reaction, DNA purification, and sample extraction) was performed within 5 min in a microchip, we could reduce the sample preparation time in comparison with that for the conventional methods used in biochemistry laboratories. Finally, we verified the performance of the sample preparation chip by conducting PCR (polymerase chain reaction) analysis of the chip product.

Effect of Red Bean Protein and Microbial Transglutaminase on Gelling Properties of Myofibrillar Protein (적소두단백질(Red Bean Protein)과 Transglutaminase를 첨가한 돈육 근원섬유 단백질의 물성 증진 효과)

  • Jang, Ho-Sik;Chin, Koo-Bok
    • Food Science of Animal Resources
    • /
    • v.31 no.5
    • /
    • pp.782-790
    • /
    • 2011
  • The effects of soy protein isolate (SPI) and red bean protein isolate (RBPI) on gelling properties of pork myofibrillar protein (MP) in the presence of microbial transglutaminase (MTG) were studied at 0.45 M NaCl. MP paste was incubated with MTG (0.1%) at various levels (0.1, 0.3, 0.5, and 1%) of SPI and RBPI before incubating at $4^{\circ}C$ for 4 h. The rheological property results showed that MP gel shear stress increased with increasing RBPI concentration. Cooking yield (CY) of the MP gel increased with increasing RBPI and SPI, whereas gel strength (GS) was not affected by adding RBPI or SPI. Thus, effects of incubation time (0, 4, 8, 10, and 12 h) were measured at 0.1% SPI and RBPI. GS values of the MP gel at 10 and 12 h were similar and were higher than those of the others. CY values were highest when RBPI (0.1%) was added, regardless of incubation time. The protein patterns indicated that incubating the MP with MTG for 10 h resulted in protein crosslinking between MP and RBPI or SPI. Based on these results, RBPI and SPI could be used as an ingredient to increase textural properties and cooking yield of meat protein gel.