• Title/Summary/Keyword: heat-denaturation

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Effects of Heat Shock Treatment on Enzymatic Proteolysis for LC-MS/MS Quantitative Proteome Analysis

  • Arul, Albert-Baskar;Han, Na-Young;Jang, Young-Su;Kim, Hyojin;Kim, Hwan-Mook;Lee, Hookeun
    • Mass Spectrometry Letters
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    • v.7 no.1
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    • pp.1-11
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    • 2016
  • Various efforts have been developed to improve sample preparation steps, which strongly depend on hands-on processes for accurate and sensitive quantitative proteome analysis. In this study, we carried out heating the sample prior to trypsin digestion using an instrument to improve the tryptic digestion process. The heat shock generated by the system efficiently denatured proteins in the sample and increased the reproducibility in quantitative proteomics based on peptide abundance measurements. To demonstrate the effectiveness of the protocol, three cell lines (A human lung cancer cell line (A549), a human embryonic kidney cell line (HEK293T), and a human colorectal cancer cell line (HCT-116)) were selected and the effect of heat shock was compared to that of normal tryptic digestion processes. The tryptic digests were desalted and analysed by LC-MS/MS, the results showed 57 and 36% increase in the number of identified unique peptides and proteins, respectively, than conventional digestion. Heat shock treated samples showed higher numbers of shorter peptides and peptides with low inter-sample variation among triplicate runs. Quantitative LC-MS/MS analysis of heat shock treated sample yielded peptides with smaller relative error percentage for the triplicate run when the peak areas were compared. Exposure of heat-shock to proteomic samples prior to proteolysis in conventional digestion process can increase the digestion efficiency of trypsin resulting in production of increased number of peptides eventually leading to higher proteome coverage.

Studies on Heat Stability of Egg Albumen Gel III. Changes of Heat Stability of Egg Albumen Gel by Chemical Modification (난백겔의 열안정성에 관한 연구 III. 화학변형에 의한 난백겔의 열안정성 변화)

  • 유익종;김창한;한석현;송계원
    • Korean Journal of Poultry Science
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    • v.17 no.3
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    • pp.217-223
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    • 1990
  • This study was undertaken to find out the changes of chemically modified egg albumen grl after heat treatment at $95^{\circ}C$ for 30 minutes or at $120^{\circ}C$ for 30 minutes. Acetylation and succinylation increased the hardness of egg albumen gel, it was rather higher at high heat treatment($120^{\circ}C$, 30min.) than at low heat treatment($95^{\circ}C$, 30min). The cohesiveness of egg albumen gel was improved remarkably by succinylation and maleylation at both low and high heat treatment. The lightness and yellowness of egg albumen gel were decreased by chemical modification. Initial heat denaturation temperature of egg albumen was increased by 11$^{\circ}C$ by acetylation, by $12.5^{\circ}C$ by maleylation and by ,$14.5^{\circ}C$ by succinylation.

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Changes in expression of monocarboxylate transporters, heat shock proteins and meat quality of Large White Yorkshire and Ghungroo pigs during hot summer period

  • Parkunan, Thulasiraman;Das, Arun K.;Banerjee, Dipak;Mohanty, Niharika;Paul, Avishek;Nanda, P.K.;Biswas, TK;Naskar, Syamal;Bag, Sadhan;Sarkar, Mihir;Mohan, Narayana H.;Das, Bikash Chandra
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.2
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    • pp.246-253
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    • 2017
  • Objective: Present study explores the effect of hot summer period on the glycolytic rate of early post-mortem meat quality of Ghungroo and Large White Yorkshire (LWY) pig and comparative adaptability to high temperature between above breeds by shifting the expression of stress related genes like mono-carboxylate transporters (MCTs) and heat shock proteins (HSPs). Methods: Healthy pigs of two different breeds, viz., LYW and Ghungroo (20 from each) were maintained during hot summer period (May to June) with a mean temperature of about $38^{\circ}C$. The pigs were slaughtered and meat samples from the longissimus dorsi (LD) muscles were analyzed for pH, glycogen and lactate content and mRNA expression. Following 24 h of chilling, LD muscle was also taken from the carcasses to evaluate protein solubility and different meat quality measurements. Results: LWY exhibited significantly (p<0.01) higher plasma cortisol and lactate dehydrogenase concentration than Ghungroo indicating their higher sensitivity to high temperature. LD muscle from LWY pigs revealed lower initial and ultimate pH values and higher drip loss compared to Ghungroo, indicating a faster rate of pH fall. LD muscle of Ghungroo had significantly lower lactate content at 45 min postmortem indicating normal postmortem glycolysis and much slower glycolytic rate at early postmortem. LD muscle of LWY showed rapid postmortem glycolysis, higher drip loss and higher degrees of protein denaturation. Ghungroo exhibited slightly better water holding capacity, lower cooking loss and higher protein solubility. All HSPs (HSP27, HSP70, and HSP90) and MCTs (MCT1, MCT2, and MCT4) in the LD muscle of pigs inclined to increase more in Ghungroo than LWY when exposed to high temperature. Conclusion: Effect of high temperature on the variation of HSPs and MCTs may play a crucial role in thermal tolerance and adaptation to different climatic conditions, pH regulation, muscle acidification, drip loss, protein denaturation and also in postmortem meat quality development.

Denaturation of Heat Treated Lysozyme under High Pressure Conditions (고압조건에서 가열 처리된 Iysozyme의 변성)

  • Cho, Rae-Kwang;Hong, Jin-Hwan
    • Korean Journal of Food Science and Technology
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    • v.23 no.3
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    • pp.366-369
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    • 1991
  • In order to elucidate texturization mechanism of extrudated protein, egg white lysozyme was heated under high pressure conditions, and its solubility and changes of molecular weight were investigated. Under high pressure conditions of $100,\;300\;and\;600\;kg/cm^2$, solubility decreased gradually with increasing temperature in the samples heated at $70,\;120\;and\;150^{\circ}C$ and decreased notably with increasing pressure at $200^{\circ}C$. Polymerization was found in the samples heated at $150\;and\;200^{\circ}C$ while a band which located below monomer(low-molecular) could be recognized. Molecular weight of the low-molecular was estimated to be about $6,000{\sim}9,000$ and no smaller peptide was recognized. The polymerization may have occured by disulfide crosslinking in the samples heated at $120^{\circ}C$ but other crosslinking may have played a role in those at $150\;and\;200^{\circ}C$.

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Improved Functional Characteristics of Whey Protein Hydrolysates in Food Industry

  • Jeewanthi, Renda Kankanamge Chaturika;Lee, Na-Kyoung;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.35 no.3
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    • pp.350-359
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    • 2015
  • This review focuses on the enhanced functional characteristics of enzymatic hydrolysates of whey proteins (WPHs) in food applications compared to intact whey proteins (WPs). WPs are applied in foods as whey protein concentrates (WPCs), whey protein isolates (WPIs), and WPHs. WPs are byproducts of cheese production, used in a wide range of food applications due to their nutritional validity, functional activities, and cost effectiveness. Enzymatic hydrolysis yields improved functional and nutritional benefits in contrast to heat denaturation or native applications. WPHs improve solubility over a wide range of pH, create viscosity through water binding, and promote cohesion, adhesion, and elasticity. WPHs form stronger but more flexible edible films than WPC or WPI. WPHs enhance emulsification, bind fat, and facilitate whipping, compared to intact WPs. Extensive hydrolyzed WPHs with proper heat applications are the best emulsifiers and addition of polysaccharides improves the emulsification ability of WPHs. Also, WPHs improve the sensorial properties like color, flavor, and texture but impart a bitter taste in case where extensive hydrolysis (degree of hydrolysis greater than 8%). It is important to consider the type of enzyme, hydrolysis conditions, and WPHs production method based on the nature of food application.

Screening on the Anti-inflammatory Activity of Crude Drugs (생약의 소염활성에 관한 검색)

  • Han, Byung-Hoon;Chi, Hyung-Joon;Han, Yong-Nam;Ryu, Kyung-Soo
    • Korean Journal of Pharmacognosy
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    • v.3 no.4
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    • pp.205-209
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    • 1972
  • Anti-inflammatory activity of crude drugs was evaluated by the albumin stabilizing activity test, according to the screening method of Mizushima et al., upon the randomly selected samples of 63 genus, 106 families, 123 species of plant. Almost every plant belonging to the families Araliaceae, Umbelliferae and Liliaceae showed strong stabilizing activity on the heat denaturation of bovine serum albumin, suggesting the presence of anti-inflammatory components in the plants.

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Studies on the Analgesic and Anti-inflammatory Effects of Youngsunjetong-Eum (영선제통음(靈仙除痛飮)이 진통(鎭痛) 및 소염작용(消炎作用)에 미치는 영향(影響))

  • Ahn, Dug-Kyun
    • Korean Journal of Pharmacognosy
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    • v.12 no.1
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    • pp.44-50
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    • 1981
  • The results of the comparison among the effects of decoction, extract powder and Gamiyoungsunjetongeum are as follows: In the experiment of analgesic effect in rats, having pain caused by Acetic acid, decoction showed a considerable reduction; extract powder did not show any notability and the effect of added decoction was conspicuous. The Anti-inflammatory effect of decoction was notable during $90{\sim}240minutes$, but extract powder also did not show any notability and added decoction showed a remarkable effect. In the experiment of inhibition on heat denaturation of serum rats, decoction and extract powder showed the inhibiting effects of 18.2%, 17.9% and the added decoction was even more remarkable(28.3%).

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Effect of Bicarbonate and Phosphate Buffer Treatments on the Structure and Thermal Stability of Spent Layer Meat (중 탄산 및 인산염 완층액 처리가 노계육의 조직구조 및 열안정성에 미치는 영향)

  • Yi, Song-Sop;Mast, Morris G.
    • Korean Journal of Food Science and Technology
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    • v.23 no.6
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    • pp.695-701
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    • 1991
  • Spent layer breast meat and leg meat samples washed with 0.05 M sodium bicarbonate solution and 0.04 M phosphate buffer(pH 8.3) showed decreases in heat denaturation temperature indicating the destabilization of myofibrillar proteins. The destabilization was attributed to the solubilization of 95 Kdalton and 55 kdalton proteins from the myofibrils observed in gel-electrophoretograms. Transmission electron microscopy further indicated the breakage of Z-lines.

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A Mueller Matrix Study for Measuring Thermal Damage Levels of Collagenous Tissues

  • Jun, Jae-Hoon
    • Journal of Biomedical Engineering Research
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    • v.27 no.6
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    • pp.310-317
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    • 2006
  • Extensive research with polarimetry and Mueller matrix has been done for chemical measurements and possible cancer detection. However, the effect of thermally denatured biological tissue on polarization changes is not well known. The purpose of this study is to characterize polarization changes in collagen due to thermal denaturation. The variations in polarized state caused by thermal damage were investigated by obtaining the Mueller matrix elements of collagen sample at multiple thermal damage levels. The changes in birefringence of denatured collagen were also investigated. This information could be used to determine the extent of thermal damage level of clinically heat treated tissues.

A Review on the Change of Physicochemical Quality during Heating of Milk (열처리에 의한 우유의 이화학적 품질변화에 관한 고찰)

  • Jung, In-Gyeong;In, Yeong-Min
    • Journal of Dairy Science and Biotechnology
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    • v.19 no.1
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    • pp.13-21
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    • 2001
  • Milk can be regarded as a complete food, containing protein, fat, lactose, vitamins and minerals. Milk is heated for a variety of reasons. The main reasons are: to remove pathogenic organisms; to increase shelf-life. But, when milk is heated, many changes take place: denaturation of whey proteins and interaction with casein, Maillard browning, losses of vitamin and minerals. The addition of a additive and milk powder to flavor and taste may cause undesirable change of quality during heating milk. The reconstituted milk is the milk product resulting from the addition of water to the dried or condensed form in the amount necessary to re-establish the specified water solids ratio. Therefore, according to the increasement of consumption of processed milk, the necessity for study about the quality of processed milk mixed with reconstituted milk arose.

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