• 제목/요약/키워드: thermal denaturation

검색결과 80건 처리시간 0.026초

"OPEN" STRUCTURE OF SecA PROTEIN OF ESCHERICHIA COLI IN SOLUTION

  • Maengseok Song;Kim, Hyounman
    • 한국생물물리학회:학술대회논문집
    • /
    • 한국생물물리학회 1996년도 정기총회 및 학술발표회
    • /
    • pp.27-27
    • /
    • 1996
  • SecA protein which has a pivotal role in the preprotein cranslocation across the inner membrane of Escherichia coli is a water-soluble protein with an unusual property of penetrating the membrane readily. An interesting and important question is what structural characteristics of SecA enables its ready penetration of lipid bilayer. The conformational properties of SecA in solution at 3$0^{\circ}C$, pH 7.5 were observed by hydrogen-tritium (HT) exchange, and denaturant-induced denaturation/renaturation and thermal unfolding. (omitted)

  • PDF

Fabrication, Estimation and Trypsin Digestion Experiment of the Thermally Isolated Micro Teactor for Bio-chemical Reaction

  • Sim, Tae-Seok;Kim, Dae-Weon;Kim, Eun-Mi;Joo, Hwang-Soo;Lee, Kook-Nyung;Kim, Byung-Gee;Kim, Yong-Hyup;Kim, Yong-Kweon
    • JSTS:Journal of Semiconductor Technology and Science
    • /
    • 제5권3호
    • /
    • pp.149-158
    • /
    • 2005
  • This paper describes design, fabrication, and application of the silicon based temperature controllable micro reactor. In order to achieve fast temperature variation and low energy consumption, reaction chamber of the micro reactor was thermally isolated by etching the highly conductive silicon around the reaction chamber. Compared with the model not having thermally isolated structure, the thermally isolated micro reactor showed enhanced thermal performances such as fast temperature variation and low energy consumption. The performance enhancements of the micro reactor due to etched holes were verified by thermal experiment and numerical analysis. Regarding to 42 percents reduction of the thermal mass achieved by the etched holes, approximately 4 times faster thermal variation and 5 times smaller energy consumption were acquired. The total size of the fabricated micro reactor was $37{\times}30{\times}1mm^{3}$. Microchannel and reaction chamber were formed on the silicon substrate. The openings of channel and chamber were covered by the glass substrate. The Pt electrodes for heater and sensor are fabricated on the backside of silicon substrate below the reaction chamber. The dimension of channel cross section was $200{\times}100{\mu}m^{2}$. The volume of reaction chamber was $4{\mu}l$. The temperature of the micro reactor was controlled and measured simultaneously with NI DAQ PCI-MIO-16E-l board and LabVIEW program. Finally, the fabricated micro reactor and the temperature control system were applied to the thermal denaturation and the trypsin digestion of protein. BSA(bovine serum albumin) was chosen for the test sample. It was successfully shown that BSA was successfully denatured at $75^{\circ}C$ for 1 min and digested by trypsin at $37^{\circ}C$ for 10 min.

말쥐치 근원섬유단백질의 열안정성과 및 가지 첨가제의 영향 (Thermal-Denaturation of File Fish Myofibrillar Protein and Protective Effect of Sucrose, Sorbitol and Amino Acids)

  • 최영준;변재형
    • 한국수산과학회지
    • /
    • 제18권5호
    • /
    • pp.455-463
    • /
    • 1985
  • 말쥐치 육의 근원섬유단백질을 추출하고 몇가지 온도조건이 그 변성에 미치는 영향을 실험하였으며, sucrose와 sorbitol 및 아미노산을 첨가하였을 때의 변성억제효과에 관하여도 비교 검토하였다. 실험에서 얻은 결과를 요약하면 다음과 같다. 1. 말쥐치 근원섬유단백질의 $25^{\circ}C,\;30^{\circ}C,\;35^{\circ}C$에서의 변성속도정수($K_D$값)는 각각 $19.52{\times}10^{-5},\;112.25{\times}10^{-5},\;247.20{\times}10^{-5}$이었다. 활성화 에너지${\Delta}E$값은 43 kcal/mole, 활성화엔탈피 ${\Delta}H$는 42.4kcal/mole, 활성화 엔트로피 ${\Delta}S$는 66.79 e.u., 그리고 자유에너지 ${\Delta}G$는 22.4kcal/mole이었다. 2. 말쥐치 근원섬유단백질이 $0^{\circ}C$에서 96시간 경과했을 때는 최초 활성의 $53\%$가 감소하였고, $-20^{\circ}C$에서 60시간이 경과했을 때는 약 $72\%$가 감소했다. 3. 농도별 첨가제의 가열변성에 대한 억제 효과는 1M Na-Glu>1M sorbitol>1.25M Gly>0.5M sucrose>1.25M Ala의 순이며, 변성억제효과 값 ${\Delta}E$는 Na-Glu가 1.25, sorbitol이 0.58, Gly이 0.28, sucrose가 0.21, Ala이 0.05이였다. 4. 혼성 첨가시는 1M Na-Glu+1.25M Gly>1M sorbitol+1.25 M Gly>1M sorbito1+0.5M sucrose>1M sorbito1+1M Na-Glu순으로 가열변성억제효과를 가지고 있으며, 이들의 $K_D$값은 위의 순서대로 $25^{\circ}C$에서 각각 $1.38{\times}10^{-5},\;11.75{\times}10^{-5},\;14.95{\times}10^{-5},\;15.66{\times}10^{-5}$으로서 무첨가시보다 $7{\sim}l.2$배의 효과를 나타내고 있었다. 5. 혼성 첨가시의 열역학적 함수로서 활성화 에너지, 활성화 엔탈피, 활성화 엔트로피, 자유 에너지는 Na-Glu+Gly 일 때, 68.4kcal/mole, 67.8 kcal/mole, 146.70 e. u., 24kca1/mole이며, sorbitol+Gly은 44kcal/mole, 43.4kcal/mole, 69.14 e. u., 22.8kcal/mole, sorbitol+sucrose는 42.7kcal/mole, 42.1kcal/mole 65.26 e. u., 22.6kcal/mole이고. sorbitol+Na-Glu는 49kcal/mole, 48.4kcal/mole, 86.49 e. u., 22.6kcal/mole이었다. 6. 냉동중의 변성억제효과는 1M Na-Glu+1.25M Gly/1M sorbito1+0.5M sucrose>1M sorbitol+1.25M $Gly{\sim}lM$ sorbito1+1M Na-Glu의 순으로 좋았다.

  • PDF

고추의 게놈 분석 (Analysis of Red Pepper (Capsicum annuum) Genome)

  • 안정선
    • Journal of Plant Biology
    • /
    • 제39권1호
    • /
    • pp.57-61
    • /
    • 1996
  • Thermal denaturation, reassociation kinectics 및 핵부피 측정 방법으로 고추 게놈의 염기 조성, kinetic component 및 게놈 크기를 조사하였다. 변성온도(Tm)에 근거한 고추 게놈의 염기 조성은 37% (G+C)였으며, Cot 커브에 근거한 고추 게놈은 카피수가 10,754, 178 및 1이고 kinetic complexity가 $5.6{\times}10^{3}\;bp,\;1.9{\times}10^{6}\;bp\;및\;1.9{\times}10^{8}\;bp$인 3개의 성분이 게놈의 4.2%, 26% 및 65%를 차지하고 있었다. 이로부터 계산한 고추 게놈의 크기는 $1.25{\times}10^{9}\;bp/C$인데, 이는 핵부피($62.4\;\mu\textrm{m}^3/C$)로부터 계산한 $4.053{\times}10^{9}\;bp/C$의 약 33%에 해당한다.

  • PDF

Comparative Study on the Postmortem Proteolysis and Shear Force during Aging of Pork and Beef Semitendinosus Muscles

  • Seokhee Han;Kyung Jo;Seul-Ki-Chan Jeong;Hayeon Jeon;Soeun Kim;Minkyung Woo;Samooel Jung;Seonmin Lee
    • 한국축산식품학회지
    • /
    • 제44권5호
    • /
    • pp.1055-1068
    • /
    • 2024
  • The differences in the proteolytic patterns and shear force of pork and beef during aging were evaluated. Pork and beef semitendinosus muscles were obtained at 24 and 48 h postmortem, respectively, and aged at 4℃ for 0 (Day 0), 7 (Day 7), and 14 days (Day 14). Changes in the electrical conductivity were observed in pork on Day 7 and beef on Day 14. The calpain activity increased in pork (p<0.05) after 14 days of aging, whereas that of beef decreased on Day 7 (p<0.05). The cathepsin B activity in pork and beef increased between Day 7 and 14 (p<0.05). The content of α-amino group in the 10% trichloroacetic acid-soluble fraction increased between Day 7 and 14 in pork (p<0.05), but increased steadily in beef throughout aging (p<0.05). The electrophoretogram of the myofibrillar proteins revealed a 30 kDa protein band only in the beef lane on Day 14. The cooked pork had no significant changes in the shear force during aging periods (p>0.05), while the gradual decrease in the shear force with the increasing aging periods was shown in the cooked beef (p<0.05). Circular dichroism analysis of myosin extracts from pork and beef revealed thermal denaturation temperatures of 55℃ and 58℃, respectively. This study highlights the different post-mortem proteolytic patterns and thermal denaturation temperatures of myosin in pork and beef semitendinosus muscles, which contribute to distinct changes in the shear force during aging between pork and beef.

랩온어칩을 위한 중합효소 연쇄반응 칩의 열설계 (Thermal Design of PCR Chip for LOC)

  • 김덕종;김재윤;박상진;허필우;윤의수
    • 연구논문집
    • /
    • 통권33호
    • /
    • pp.17-25
    • /
    • 2003
  • In this work, thermal design of a PCR chip for LOC is systematically conducted. From the numerical simulation of a PCR chip based on the finite volume method, how to control the average temperature of a PCR chip and the temperature difference between the denaturation zone and the annealing zone is presented. The average temperature is shown to be controlled by adjusting heat input and a cooler as well as a heater is shown to be necessary to obtain three individual temperature zones for polymerase chain reaction. To reduce the time required, a heat sink for the cooler is not included in the calculation domain for the PCR chip and heat sink design is conducted separately by using a compact modeling method, the porous medium approach.

  • PDF

Screening Molecular Chaperones Similar to Small Heat Shock Proteins in Schizosaccharomyces pombe

  • Han, Jiyoung;Kim, Kanghwa;Lee, Songmi
    • Mycobiology
    • /
    • 제43권3호
    • /
    • pp.272-279
    • /
    • 2015
  • To screen molecular chaperones similar to small heat shock proteins (sHsps), but without ${\alpha}$-crystalline domain, heat-stable proteins from Schizosaccharomyces pombe were analyzed by 2-dimensional electrophoresis and matrix assisted laser desorption/ionization time-of-flight mass spectrometry. Sixteen proteins were identified, and four recombinant proteins, including cofilin, NTF2, pyridoxin biosynthesis protein (Snz1) and Wos2 that has an ${\alpha}$-crystalline domain, were purified. Among these proteins, only Snz1 showed the anti-aggregation activity against thermal denaturation of citrate synthase. However, pre-heating of NTF2 and Wos2 at $70^{\circ}C$ for 30 min, efficiently prevented thermal aggregation of citrate synthase. These results indicate that Snz1 and NTF2 possess molecular chaperone activity similar to sHsps, even though there is no ${\alpha}$-crystalline domain in their sequences.

Fabrication of a polymerase chain reaction micro-reactor using infrared heating

  • Im, Ki-Sik;Eun, Duk-Soo;Kong, Seong-Ho;Shin, Jang-Kyoo;Lee, Jong-Hyun
    • 센서학회지
    • /
    • 제14권5호
    • /
    • pp.337-342
    • /
    • 2005
  • A silicon-based micro-reactor to amplify small amount of deoxyribonucleic acid (DNA) has been fabricated using micro-electro-mechanical systems (MEMS) technology. Polymerase chain reaction (PCR) of DNA requires a precise and rapid temperature control. A Pt sensor is integrated directly in the chamber for real-time temperature measurement and an infrared lamp is used as external heating source for non-contact and rapid heating. In addition to the real-time temperature sensing, PCR needs a rapid thermocycling for effective PCR. For a fast thermal response, the thermal mass of the reactor chamber is minimized by removal of bulk silicon volume around the reactor using double-side KOH etching. The transparent optical property of silicon in the infrared wavelength range provides an efficient absorption of thermal energy into the reacting sample without being absorbed by silicon reactor chamber. It is confirmed that the fabricated micro-reactor could be heated up in less than 30 sec to the denaturation temperature by the external infrared lamp and cooled down in 30 sec to the annealing temperature by passive cooling.

Temperature Distributions of the Lumbar Intervertebral Disc during Laser Annuloplasty : A Cadaveric Study

  • Lee, Min Hyung;Kim, Il Sup;Hong, Jae Taek;Sung, Jae Hoon;Lee, Sang Won;Kim, Daniel H.
    • Journal of Korean Neurosurgical Society
    • /
    • 제59권6호
    • /
    • pp.559-563
    • /
    • 2016
  • Objective : Low back pain, caused intervertebral disc degeneration has been treated by thermal annuloplasty procedure, which is a non-surgical treatement. The theoretical backgrounds of the annuloplasty are thermal destruct of nociceptor and denaturization of collagen fiber to induce contraction, to shrink annulus and thus enhancing stability. This study is about temperature and its distribution during thermal annuloplasty using 1414 nm Nd : YAG laser. Methods : Thermal annuloplasty was performed on fresh human cadaveric lumbar spine with 20 intact intervertebral discs in a $37^{\circ}C$ circulating water bath using newly developed 1414 nm Nd : YAG laser. Five thermocouples were attached to different locations on the disc, and at the same time, temperature during annuloplasty was measured and analyzed. Results : Thermal probe's temperature was higher in locations closer to laser fiber tip and on lateral locations, rather than the in depth locations. In accordance with the laser fiber tip and the depth, temperatures above $45.0^{\circ}C$ was measured in 3.0 mm depth which trigger nociceptive ablation in 16 levels (80%), in accordance with the laser fiber end tip and laterality, every measurement had above $45.0^{\circ}C$, and also was measured temperature over $60.0^{\circ}C$, which can trigger collagen denaturation at 16 levels (80%). Conclusion : When thermal annuloplasty is needed in a selective lesion, annuloplasty using a 1414 nm Nd : YAG laser can be one of the treatment options.

pH, NaCl 및 phosphate 첨가에 따른 돈육 드립의 열안정성 및 색에 관한 연구 (Studies on the Thermal Stability and Color of Free Drip released from Pork Muscle with pH, Concentration of NaCl and Phosphate)

  • 김천제;이창현;송민석;이의수;조진국
    • 한국식품과학회지
    • /
    • 제32권6호
    • /
    • pp.1285-1290
    • /
    • 2000
  • 본 실험은 수소이온 농도, 염, Sodium tripolyphosphate(STPP) 농도에 따른 돈육 등심 드립의 열변성을 DSC를 이용하여 조사한 것이다. pH가 증가함에 따라 돈육 등심 드립의 열안정성은 증가하였으며 특히 $T_1$$pH\;5.5{\sim}6.0$에서 $6.33^{\circ}C$의 가장 큰 증가를 보였다. 0.1 N, 0.2 N NaOH/1, 2 N HCl 및 $0{\sim}5%$ NaCl과 $0{\sim}0.5%$ sodium tripolyphosphate(STPP)로 처리하였을 때, pH가 증가함에 따라 드립의 열안정성은 증가하였으며, 특히 $T_1$$pH\;5.5{\sim}6.0$에서 $6.33^{\circ}C$의 가장 큰 증가를 보였다. 그리고 NaCl농도의 증가는 드립의 열안정성을 감소시켰는데 그중 $T_1$은 5% NaCl농도 수준에서 $9.41^{\circ}C$의 감소를 보였다. 한편 STPP는 드립이 열안정성을 증가시켰으며 0.5% STPP농도 수준에서 $T_1$$5.84^{\circ}C$의 증가를 보였다. $40{\sim}100^{\circ}C$로 증가함에 따라 $Lightness(L^*)$는 41.13에서 69.54로 증가하였고 $redness(a^*)$는 26.70에서 5.40으로 감소하였다. 특히 $L^*-$$a^*-$값 모두 $40{\sim}60^{\circ}C$에서 약 78%의 가장 큰 변화를 보였다.

  • PDF