• Title/Summary/Keyword: Protein denaturation

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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.

Efficient Diagnosis of Cucumber Green Mottle Mosaic Virus in Watermelon Using RT-PCR and Cloning of Coat Protein Gene (RT-PCR을 이용한 수박 Cucumber Green Mottle Mosaic Virus의 효율적인 진단 및 외피단백질 유전자의 클로닝)

  • 양덕춘;이진숙;김두욱;임용표;민병훈
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.6
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    • pp.519-524
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    • 1998
  • A simple and reliable method to diagnose cucumber green mottle mosaic virus of watermelon in Korea (CGMMV-WK) was determined by RT-PCR, and coat protein gene for CGMMV-WK was cloned. Comparing to a method reported by Lee et al. (1996), the method developed here showed a better RT-PCR reaction. RT-PCR was possible by one step in the PCR reaction mixture that contains 20 pmol of primer, reverse transcriptase (30 unit), RNasin (5 unit) using the crude RNA solution. RT-PCR condition for specifically diagnosing CGMMV-WK was that cDNA was synthesized at 42$^{\circ}C$ for 45 min followed by pre-denaturation at 95$^{\circ}C$ for 2 min, and then PCR reaction was carried out with a programmed condition that consisted of 36 sequential cycles at 96$^{\circ}C$ for 30 sec, 6$0^{\circ}C$ for 30 sec, and 72$^{\circ}C$ for 1 min. A gene encoding the coat protein of CGMMV-WK was cloned and characterized. Nucleotide sequence of coat protein gene of CGMMV-WK shared 98.77% and 99.38% of sequence identity with those of CGMMV-W and CGMMV-SH, respecitvely, however, all of amino acid sequences were same.

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The changes of the plasma protein and the complements ($C_{3}$, $C_{4}$) after open heart surgery (개심수술후 혈장 단백 및 보체 ($C_{3}$, $C_{4}$)의 변화상 추적)

  • 남충희
    • Journal of Chest Surgery
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    • v.19 no.4
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    • pp.558-562
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    • 1986
  • The extracorporeal circulation has been much improved recently, but has yet much complex problems such as the protein denaturation and the activation of the complement system by the exposure of the blood to the foreign surface, which may result in such as the postperfusion syndrome. We studied the changes of the plasma protein fractions by the electrophoresis and the complement consumption [C3, C4] by the immunodiffusion method in the patients undergoing cardiac operation from Mar. 1, 1986 to Aug. 31, 1986. The results were summarized as follows: 1. y-globulin fraction was decreased [p<0.02 by paired t-test, N=25], but a,-globulin was increased [p<0.001 by paired t test, N=25] after operation. 2. C3,C4 were significantly reduced [p<0.001 by paired t-test, N=14] postoperatively and normalized from 24 hours after operation. 3. The consumption of C3,C4 had significant linear correlation [correlation coefficient r=0.97] and C, was more markedly reduced comparing with C3, which probably means the complement activation by classical pathway in our bubble oxygenator group.

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Chemical Quality Evaluation of Commercial Infant Formulas (시판 조제 분유의 화학적 품질 평가)

  • 한정은;홍윤호
    • Food Science of Animal Resources
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    • v.21 no.1
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    • pp.56-63
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    • 2001
  • In this study, three domestic and one foreign formulas for the infants up to 5 month old were examined to detect chemical changes such as pH, reactive sulfhydryl groups(RSH) content, 5-hydroxymethylfurfral(HMF) content, available lysine content, electrophoresis, and surface color caused by heat treatment for long term storage. In the SDS-polyacrylamide gel electrophoresis, A and B products showed similar pattern, while C product had a clearly distinguishable $\beta$-lacto-globulin band, but in casein, only D product showed a few strong casein band. RSH content, which indicate the extent of whey protein denaturation, ranged from 4.40 to 5.93 mmole/g protein. HMF content. which indicate the extent of Maillard reaction, ranged from 192 to 432 $\mu$mole/100g in formulas. B product showed the highest RSH and HMF content. Available lysine content ranged from 31 to 46 mg/g protein. Among them D product contain the highest available lysine content and others showed no significant difference. In conclusion, the domestic infant formulas showed higher RSH and HMF content than the foreign product and the available lysine content of the domestic products were lower than of the foreign product.

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The Disruption of Saccharomyces cerevisiae Cells and Release of Glucose 6-Phosphate Dehydrogenase (G6PDH) in a Horizontal Dyno Bead Mill Operated in Continuous Recycling Mode

  • Mei Chow Yen;Ti Tey Beng;Ibrahim Mohammad Nordin;Ariff Arbakariya;Chuan Ling Tau
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.3
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    • pp.284-288
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    • 2005
  • Baker's yeast was disrupted in a 1.4-L stainless steel horizontal bead mill under a continuous recycle mode using 0.3 mm diameter zirconia beads as abrasive. A single pass in continuous mode bead mill operation liberates half of the maximally released protein. The maximum total protein release can only be achieved after passaging the cells 5 times through the disruption chamber. The degree of cell disruption was increased with the increase in feeding rate, but the total protein release was highest at the middle range of feeding rate (45 L/h). The total protein release was increased with an increase in biomass concentration from 10 to $50\%$(w/v). However, higher heat dissipation as a result of high viscosity of concentrated biomass led to the denaturation of labile protein such as glucose 6-phosphate dehydrogenase (G6PDH). As a result the highest specific activity of G6PDH was achieved at biomass concentration of $20\%$(ww/v). Generally, the degree of cell disruption and total protein released were increased with an increase in impeller tip speed, but the specific activity of G6PDH was decreased substantially at higher impeller tip speed (14 m/s). Both the degree of cell disruption and total protein release increased, as the bead loading increased from 75 to $85\% (v/v)$. Hence, in order to obtain a higher yield of labile protein such as G6PDH, the yeast cell should not be disrupted at biomass concentration and impeller tip speed higher than $20\%(w/v)$ and 10 m/s, respectively.

Effect of Lipid on the Protein Denaturation in Cooking Fish Meat (어육 조리시 지질이 단백질 변성에 미치는 영향)

  • LEE Kang-Ho;CHOI Byeong-Dae;RYU Hong-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.5
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    • pp.493-501
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    • 1986
  • The present work aims to estimate the effect of heat treatment on the in vitro protein digestibility and formation of trypsin inhibitor or trypsin indigestible substrate(TIS) of raw and defatted flounder. It was also carried out to assess the formation of lipid-protein complexes under the conditions of different ratio of lipid addition. The in vitro protein digestibility increased when steamed for 5 min showing $88.09\%$ in raw and $90.56\%$ in defatted samples, respectively. After 40 min steaming, the digestibility decreased by $2{\sim}4\%$. As for microwaving, heating for 1 min resulted in slight increase of digestibility, however, heating for 7 min did decrease of digestibility by $3{\sim}4\%$ for both raw and defatted materials. There was no difference in fatty acid composition found with heat treatment. The major fatty acids of flounder meat were $C_{16:0},\;C_{16:1},\;C_{18:1},\;C_{20:5},\;C_{22:6}$ and the ratio of the unsaturated to saturated was 67.3:32.6. Fat oxidation and nonenzymatic browning were enhanced by heat treatment and protein solubility decreased necessarily as the brown pigment formation increased. On the other hand, the effects on the digestibility and TIS of the complexes formed from interaction of lipid and myofibrillar or meat protein of flounder were examined. The interaction of protein with lipid was considered to mostly contribute to the drop of digestibility of fish products. The digestibility of myofibrillar protein was $93.72\%$ for flounder, and it generally decreased as the amount of lipid added to protein and reaction time increased. Also mixed and heated samples were more active in digestibility decline than those mixed after heating. The result probably indicated that lipid-protein interaction was involved in the drop of digestibility which coincided with protein denaturation.

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Conformation of Soymilk Protein Treated by Pretense (단백분해효소 처리된 두유단백질의 구조적 특성)

  • 변진원
    • The Korean Journal of Food And Nutrition
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    • v.15 no.4
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    • pp.331-336
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    • 2002
  • Conformation of soymilk protein was examined to obtain basic information for improved calcium intolerence of soymilk protein partially hydrolyzed with protease. Surface hydrophobicities of three proteins showed the order of SMP(soymilk protein) < SPI(soy protein isolate) < PT-SMP(protease treated soymilk protein). Total thiol group contents of SMP and PT-SMP were similar but larger than that of SPI. Reducing rate of disulfide bond in PT-SMP after 2-mercaptoethanol treatment was laster than that in SMP. And so, this result indicates that PT-SMP may be less compacting due to protease treatement. From circular dichroism result, PT-SMP showed different pattern from SMP and SPI suggesting change of secondary structure by hydrolysis. And analysis of heat denaturating property by DSC showed that denaturation enthalpy of three proteins were all small. Especially enthalpy of PT-SMP was least, and this result suggested that PT-SMP was denatured easily by heating due to less compacting structure.

Microbead based micro total analysis system for Hepatitis C detection (마이크로비드를 이용한 초소형 C형 간염 검출 시스템의 제작)

  • Sim, Tae-Seok;Lee, Bo-Rahm;Lee, Sang-Myung;Kim, Min-Soo;Lee, Yoon-Sik;Kim, Byung-Gee;Kim, Yong-Kweon
    • Proceedings of the KIEE Conference
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    • 2006.07c
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    • pp.1629-1630
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    • 2006
  • This paper describes a micro total analysis system ($\mu$ TAS) for detecting and digesting the target protein which includes a bead based temperature controllable microchip and computer based controllers for temperature and valve actuation. We firstly combined the temperature control function with a bead based microchip and realized the on-chip sequential reactions using two kinds of beads. The PEG-grafted bead, on which RNA aptamer was immobilized, was used for capturing and releasing the target protein. The target protein can be chosen by the type of RNA aptamer. In this paper, we used the RNA aptamer of HCV replicase. The trypsin coated bead was used for digesting the released protein prior to the matrix assisted laser desorption ionization time of flight mass spectrometer (MALDI TOF MS). Heat is applied for release of the captured protein binding on the bead, thermal denaturation and trypsin digestion. PDMS microchannel and PDMS micro pneumatic valves were also combined for the small volume liquid handling. The entire procedures for the detection and the digestion of the target protein were successfully carried out on a microchip without any other chemical treatment or off-chip handling using $20\;{\mu}l$ protein mixture within 20 min. We could acquire six matched peaks (7% sequence coverage) of HCV replicase.

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Induction of Leptin cDNA Expression in Esherichia coli Cells (대장균 세포에서 Leptin 유전자의 발현 유도)

  • 김은정;정인철;오상환;조무연
    • Journal of Life Science
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    • v.9 no.3
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    • pp.253-261
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    • 1999
  • Leptin gene, an obesity gene, has been known to involve in the regulation of food intake and body weight. It is also thought to be related to the glucose metabolism, insulin secretion and type II diabetes mellitus. Recently, the production of recombinant leptin protein has been attempted for the application in the treatment of obesity and the correction of hereditary obesity and type II diabetes. In the present study, leptin cDNA was cloned from mouse fat cells by RT-PCR and prokaryotic expression of leptin was attempted in order ot prepare a leptin-specific antigen. Immunization of a rabbit with the leptin-specific antigen into a rabbit resulted in the generation of leptin-specific antiserum that could be useful in the detection of leption expressed in various tissues. The sequence of leptin cDNA prepared in the present study wa identical to the previously reported one. Transformation of E. coli(DH5a) cells with the leptin cDNA-inserted translation vector, pGEX-4T-3-leptin followed by treatment with IPTG (0.1mM) resulted in the expression of a large amount of GST-leptin fusion protein with a molecular weight of 44 KDa as an inclusion body. Denaturation of the insoluble fusion protein by 8M urea, 6M guanidium-HCI or 0.1% 2-mercaptoethanol followed by a slow oxidation could not solubilize the inclusion body. The cell extract was subjected to SDS-PAGE and GST-leptin protein electroeluted from the gel was then injected into a rabbit subcutaneously for the immunization. Anti-GST-leptin rabbit antiserum which had a cross reactivity to the GST-leptin protein was generated. Leptin protein expressed in mouse brain and fat tissues was detected by Western blot immunodetection system using the antiserum generated in the present study.

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Changes in the $Ca^{2+}$-, $Mg^{2+}$- dependent Adenosine Triphosphatase Activity and Ultrastructure of Marine Fishes by Partial Freezing II. Changes in ATPase Activity of Yellowtail Actomyosin during Cold Storage (해산어의 부분동결에 의한 $Ca^{2+}$-, $Mg^{2+}$- dependent Adenosine Triphosphatase 활성 및 근섬유의 미세구조 변화 II. 저온저장에 의한 방어 Actomyosin ATPase의 활성변화)

  • 박찬성;최경호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.19 no.4
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    • pp.349-355
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    • 1990
  • Actomyosine prepared from Yellowtail fish(seriola quinqueradiata) were stored at $0^{\circ}C$(ice-cooling) -3.5$^{\circ}C$(partial freeaing) and -2$0^{\circ}C$(freezing) Another actomyosin samples were prepared from the fish previously stored at the temperatures for a week as the maximum .Remaining activity of {{{{ {Ca }^{2+ } }}}}-and {{{{ {Mg }^{2+ } }}}}- dependent adenosine triphosphatase(ATPase) activity was measured fronm the actomyosin preparations. Specific activity of {{{{ {Mg }^{2+ } }}}}-ATPase of actomy-osin before storagew was 0.253$\mu$ mole pi/min/mg of protein and it was 1.5 times higher than that of {{{{ {Ca }^{2+ } }}}} -ATPase. The enzyme activities were markedly decreased during early period of storage. However no significant differences in the enzyme activity were revealed among the samples stored at different temperature. The enzyme of actomyosin prepared from the fish previously stored at the temperatures for a week revealed an acitivity of 2-3 times higher than that of freezing. Apparent denaturation constant of {{{{ {Mg }^{2+ } }}}} -ATPase of actomyosin was between 0.810-1.139 per day and it was about 1.5 times hgiher than that of {{{{ {Ca }^{2+ } }}}} -ATPase. But the constant of {{{{ {Mg }^{2+ } }}}} ATPase of actomyosin extracted from the fist stored for a week at each temperature was between 0.176-0.356 per day. This constant was 4 times higher than that of {{{{ {Ca }^{2+ } }}}}- ATPase in frozen stored fish. It was presumed from these results that denaturation of ATPase is largely accorded to the structural changes of actomyosin.

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