• Title/Summary/Keyword: thermal denaturation temperature

Search Result 46, Processing Time 0.027 seconds

Studies on the Denaturation of PSE Porcine Muscle Proteins by Differential Scanning Calorimetry (DSC를 이용한 PSE돈(豚) 육단백질(肉蛋白質)의 변성(變性)에 관한 연구(硏究))

  • Kim, Cheon-Jei;Honikel, K.O.;Choe, Byung-Kyu
    • Korean Journal of Food Science and Technology
    • /
    • v.21 no.2
    • /
    • pp.173-179
    • /
    • 1989
  • The influence of the storage temperature and time after slaughter on the thermal denaturation of PSE porcine muscle protein was studied by differential scanning calorimetry and by measuring the solubility of the sarcoplasmic proteins. In the DSC therodiagram a decrease of the endotherm enthalpy of the myosin plus sarcoplasmic proteins in PSE muscle could be observed with an increase in the storage temperature and time of post mortem. Storage temperature at $20^{\circ}C$ during the first four hours of post mortem resulted in relatively slight denaturation of myosin plus sarcoplasmic proteins in PSE muscle. Storage temperature above $25^{\circ}C$ caused to increase the denaturation of muscle proteins. The minimal drip loss in PSE muscle could be observed, when the muscle was cooled to $2^{\circ}C$ as quickly as possible post mortem. However, when stored for several hours of post morte at a temperature between $32^{\circ}C-38^{\circ}C$, the drip loss reached the level established for PSE muscle. The paleness of PSE muscle could be prevented to some extent by rapid chill to $20^{\circ}C$ post mortem. The more the muscle proteins in the PSE muscle become denatured during the early storage period of post mortem, the more the drip loss increases. With the increase in the denaturation of myosin plus sarcoplasmic proteins in PSE muscle with regard to temperature of post mortem, there was a corresponding decrease in the solubility of the sarcoplasmic proteins in PSE muscle.

  • PDF

Synthesis, Characterization and DNA Interaction Studies of (N,N'-Bis(5-phenylazosalicylaldehyde)-ethylenediamine) Cobalt(II) Complex

  • Sohrabi, Nasrin;Rasouli, Nahid;Kamkar, Mehdi
    • Bulletin of the Korean Chemical Society
    • /
    • v.35 no.8
    • /
    • pp.2523-2528
    • /
    • 2014
  • In the present study, at first, azo Schiff base ligand of (N,N'-bis(5-phenylazosalicylaldehyde)-ethylenediamine) ($H_2L$) has been synthesized by condensation reaction of 5-phenylazosalicylaldehyde and ethylenediamine in 2:1 molar ratio, respectively. Then, its cobalt complex (CoL) was synthesized by reaction of $Co(OAc)_2{\cdot}4H_2O$ with ligand ($H_2L$) in 1:1 molar ratio in ethanol solvent. This ligand and its cobalt complex containing azo functional groups were characterized using elemental analysis, $^1H$-NMR, UV-vis and IR spectroscopies. Subsequently, the interaction between native calf thymus deoxyribonucleic acid (ct-DNA) and CoL complex was investigated in 10 mM Tris/HCl buffer solution, pH = 7 using UV-vis absorption, thermal denaturation technique and viscosity measurements. From spectrophotometric titration experiments, the binding constant of CoL complex with ct-DNA was found to be $(2.4{\pm}0.2){\times}10^4M^{-1}$. The thermodynamic parameters were calculated by van't Hoff equation.The enthalpy and entropy changes were $5753.94{\pm}172.66kcal/mol$ and $43.93{\pm}1.18cal/mol{\cdot}K$ at $25^{\circ}C$, respectively. Thermal denaturation experiments represent the increasing of melting temperature of ct-DNA (about $0.93^{\circ}C$) due to binding of CoL complex. The results indicate that the process is entropy-driven and suggest that hydrophobic interactions are the main driving force for the complex formation.

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
    • /
    • v.5 no.3
    • /
    • 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.

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

  • Kim, Deok-Jong;Kim, Jae-Yun;Park, Sang-Jin;Heo, Pil-U;Yun, Ui-Su
    • 연구논문집
    • /
    • s.33
    • /
    • 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

Interaction of ct-DNA with 2,4-Dihydroxy Salophen

  • Azani, Mohammad-Reza;Hassanpour, Azin;Bordbar, Abdol-Khalegh;Mirkhani, Valiollah
    • Bulletin of the Korean Chemical Society
    • /
    • v.30 no.9
    • /
    • pp.1973-1977
    • /
    • 2009
  • In the present study, at first, 2,4-Dihydroxy Salophen (2,4-DHS), has been synthesized by combination of 1, 2-diaminobenzene and 2,4-dihydroxybenzaldehyde in a solvent system. This ligand containing meta-quinone functional groups were characterized using UV-Vis and IR spectroscopies. Subsequently, the interaction between native calf thymus deoxyribonucleic acid (ct-DNA) and 2,4-DHS, was investigated in 10 mM Tris/HCl buffer solution, pH 7.2, using UV-visible absorption and fluorescence spectroscopies, thermal denaturation technique and viscosity measurements. From spectrophotometric titration experiments, the binding constant of 2,4-DHS with ct-DNA was found to be (1.1 ${\pm}\;0.2)\;{\times}\;10^4\;M^{-1}.$ The fluorescence study represents the quenching effect of 2,4-DHS on bound ethidium bromide to DNA. The quenching process obeys linear Stern-Volmer equation in extended range of 2,4-DHS concentration. Thermal denaturation experiments represent the increasing of melting temperature of DNA (about 3.5 ${^{\circ}C}$) due to binding of 2,4-DHS. These results are consistent with a binding mode dominated by interactions with the groove of ct-DNA.

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
    • Journal of Sensor Science and Technology
    • /
    • v.14 no.5
    • /
    • 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
    • /
    • v.59 no.6
    • /
    • 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.

Enhancement of Hydroxylamine Reactivity of Bacteriorhodopsin at High Temperature

  • Sonoyama, Masashi;Mitaku, Shigeki
    • Journal of Photoscience
    • /
    • v.9 no.2
    • /
    • pp.299-301
    • /
    • 2002
  • Recent denaturation experiments of bacteriorhodopsin (bR) in the dark and under illumination at high temperatures revealed that irreversible thermal bleaching occurs above ~ 70°C and the preceding reversible structural changes in the dark above 60°C are closely related to irreversible photobleaching observed in the same temperature range (Yokoyama et al. (2002). J Biochem. 131,785). In this study, structural properties of bacteriorhodopsin (bR) at high temperatures were extensively probed by hydroxylamine reactivity with the Schiff base in the dark and hydrogen-deuterium (H-D) exchange in the peptide groups. In the Arrhenius plot from kinetics measurements of the hydroxylamine reaction, a good linear relationship between the reaction time constant and the inverse of the absolute temperature was observed below 60°C, while significant increase started above 60°C, suggesting that remarkable increase in water accessibility of the Schiff base in the temperature region. FT-IR spectroscopic studies on the H-D exchange suggested increase in the deuterium exchanges rate of the peptide hydrogen in the same temperature region.

  • PDF

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

  • 안정선
    • Journal of Plant Biology
    • /
    • v.39 no.1
    • /
    • pp.57-61
    • /
    • 1996
  • The genome of red pepper was investigated by thermal denaturation, reassociation kinetics and measurement of nuclear volume for its base composition, spectrum of kinetic components and genome size. Base composition was estimated to be 37% (G+C) based on melting temperature. The reassociation of 300 nt fragments analyzed by hydroxyapatite chromatography revealed the presence of three kinetic components differing in fraction of genome, kinetic complexity and number of copies as follows; 4.8% (fast) with $5.6{\times}10^{4}\;bp$ and 10,754, 26% (intermediate) with $1.9{\times}10^{6}\;bp$ and 177, and 65% (slow) with $8.48{\times}10^{8}\;bp$ and 1. These measurements demonstrate that the genome of red pepper has a 1C DNA content of $1.25{\times}10^{9}\;bp$, which is about 33% of $4.05{\times}10^{9}\;bp$ calculated from nuclear volume of $62.4\;\mu\textrm{m}^3/1C$..

  • PDF

Heterologous Expression of Recombinant Transglutaminase in Bacillus subtilis SCK6 with Optimized Signal Peptide and Codon, and Its Impact on Gelatin Properties

  • Wang, Shiting;Yang, Zhigang;Li, Zhenjiang;Tian, Yongqiang
    • Journal of Microbiology and Biotechnology
    • /
    • v.30 no.7
    • /
    • pp.1082-1091
    • /
    • 2020
  • Microbial transglutaminases (MTGs) are widely used in the food industry. In this study, the MTG gene of Streptomyces sp. TYQ1024 was cloned and expressed in a food-grade bacterial strain, Bacillus subtilis SCK6. Extracellular activity of the MTG after codon and signal peptide (SP Ync M) optimization was 20 times that of the pre-optimized enzyme. After purification, the molecular weight of the MTG was 38 kDa and the specific activity was 63.75 U/mg. The optimal temperature and pH for the recombinant MTG activity were 50℃ and 8.0, respectively. MTG activity increased 1.42-fold in the presence of β-ME and 1.6-fold in the presence of DTT. Moreover, 18% sodium chloride still resulted in 83% enzyme activity, which showed good salt tolerance. Cross-linking gelatin with the MTG increased the strength of gelatin 1.67 times and increased the thermal denaturation temperature from 61.8 to 75.8℃. The MTG also significantly increased the strength and thermal stability of gelatin. These characteristics demonstrated the huge commercial potential of MTG, such as for applications in salted protein foods.