• Title/Summary/Keyword: Isoelectric Point

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Molecular Cloning and Expression Analysis of Red-spotted Grouper, Epinephelus akaara Hsp70 (수온변화에 따른 붉바리(Epinephelus akaara)의 heat shock protein (Hsp) 70 mRNA 발현)

  • Min, Byung Hwa;Hur, Jun Wook;Park, Hyung Jun
    • Journal of Life Science
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    • v.28 no.6
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    • pp.639-647
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    • 2018
  • A new heat shock protein 70 was identified in red-spotted grouper (Epinephelus akaara) based on an expression analysis. The cDNA of red-spotted grouper Hsp70 (designated RgHsp70) was cloned by the rapid amplification of cDNA ends (RACE) techniques. The full-length of RgHsp70 cDNA was 2,152 bp, consisting of a 5'-terminal untranslated region (UTR) of 105 bp, a 3'-terminal UTR of 274 bp, and an open reading frame (ORF) of 1,773 bp that encode a polypeptide of 590 amino acids with a theoretical molecular weight of 64.9 kDa and an estimated isoelectric point of 5.2. Multiple alignment and phylogenetic analyses revealed that the RgHsp70 gene shares a high similarity with other Hsp70 fish genes. RgHsp70 contained all three classical Hsp70 family signatures. The results indicated the RgHsp70 is a member of the heat shock protein 70 family. RgHsp70 mRNA was predominately expressed in the liver, with reduced expression noted in the head-kidney tissues. The expression analysis of different water temperatures (21, 18, 15 and $12^{\circ}C$) for sampled livers revealed that expression gradually increased at $12^{\circ}C$ compared to $21^{\circ}C$. In this study, the effects of water temperature lowering on the physiological conditions were investigated, and the results revealed that novel RgHsp70 may be an important molecule involved in stress responses.

Factors Affecting Foam Separation of Proteins (단백질의 기포분리에 영향을 미치는 요소들에 관한 연구)

  • Lee, Boo-Young;Lee, Cherl-Ho
    • Korean Journal of Food Science and Technology
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    • v.19 no.3
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    • pp.220-224
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    • 1987
  • The concentration ranges forming surface excess of bovine serum albumin(BSA) and ovalbumin solutions were determined, and the factors affecting the foam separation of BSA were investigated. The surface tension of BSA solution decreased from 72 to 61 dynelcm, when the concentration changed from $5{\times}10^{-3}$ to $3{\times}10^{-2}%$, and the critical micelle concentration was appeared to be at 0.03% of BSA. At the isoelectric point (pH 4.9) of BSA, the foamate volume was maximum, but enrichment ratio was minimum, resulting in the maximum recovery rate. When the pH deviated from the isoelectric point, the foamate volume decreased and the enrichment ratio increased. The enrichment ratio increased, while the foamate volume decreased drastically as the temperature was elevated above $20^{\circ}C$, resulting in the decrease in recovery rate. As the gas flow rate increased, the enrichment ratio decreased and the foamate volume increased. When $(NH_4)_2SO_4$ was added, the enrichment ratio decreased, but the maximum foamate was obtained at ionic strength 7. The concentration to form the surface excess of ovlbumin, which has lower surface hydrophobicity than BSA, was 200 times higher than that of BSA. This fact indicates the possibility of selective foam separation by hydrophobicity difference of proteins.

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Recovery and Utilization of Proteins and Lipids from the Washing Wastewater in Marine Manufacture by Isoelectric Point Shifting Precipitation Method -3. Utilization of the Recovered Lipids as the Material for a Processed Food- (수산가공공장폐액의 등절점이동 응집처리에 의한 유용성불재회수이용 -3. 회수지방의 가공식품소재로서의 이용-)

  • SUH Jae-Soo;CHO Soon-Yeong;SON Kwang-Tae;LEE Eung-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.2
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    • pp.157-162
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    • 1995
  • When fish meat is washed for the processing of surimi, about 50ff of lipid in the fish meat is removed from the fish meat to the effluent. The removed lipid was easily recovered by centrifugation or filteration of wastewater washed fish meat. Then, the recovered lipid was utilized as a material of mayonnaise sauce processing. The major fatty acids in the recovered lipids are $C_{16:0},\;C_{18:0},\;C_{16:1},\;C_{20:5},\;and\;C_{22:6}$ Polyenoic fatty acids were composed of $33.6\%$ to total fatty acids. When the recovered lipid was substituted for soybean oil in processing of mayonnaise sauce, the maximum percentage of substitution ratio presumed to be $30\%$ according to viscosity, color difference, and emulsion stability evalution for the substituted ones.

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Codon usage analysis of rice glutelin genes (쌀 저장 단백질 글루텔린 유전자 암호 분석)

  • Shin, Yun-Cheol;Kim, Ju-Kon;Nahm, Baek-Hie
    • Applied Biological Chemistry
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    • v.36 no.6
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    • pp.517-524
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    • 1993
  • To characterize glutelins, the most abundant storage protein in rice, 13 complete coding sequences of glutelin genes from the database were analyzed. According to the phylogenic analysis, these genes could be classified into 5 groups, Group I to V. The degrees of homology were calculated to be in the range of 90 to 60%, but the patterns of hydrophobicity were similar in all the groups. Also, each group was found to have similar amino acid composition with variations in lysine content from 2.5 to 3.6% due to the point mutation of arginine to lysine. The isoelectric points of mature proteins and their basic chains of all the groups showed the value of about 9.0 and 10.0, respectively, while the isoelectric points of acidic chains in these groups showed the distinct value of 6.6, 6.7, 7.2, 8.4 and 7.9. The plot of the fraction of G+C at synonimous site in codons (GC3s) against effective codon numbers suggest no major difference in translational efficiency in the expression of glutelin multigenes.

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Sugar Content and Protein Fractionation in Human Pleural Fluid (늑막액의 당 및 단백분획상)

  • Kim, W.J.;Ahn, Y.S.;Kim, H.Y.;Lee, W.Y.
    • The Korean Journal of Pharmacology
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    • v.15 no.1_2 s.25
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    • pp.1-5
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    • 1979
  • Previous studies concerning the usefulness of pleural fluid glucose levels in differentiating causes of pleural effusions have been conflicting. Gelenger and Wiggers (1949), Calnan et al(1951) and Barber et al(1957) concluded that the lower the level of pleural fluid glucose, the more likely was tuberculosis, and that tuberculosis was unlikely if the pleural fluid glucose level was more than 80 mg/100 ml. Light and Ball(1973), however, reported that in the great majority of tuberculous pleural fluids the glucose concentration was high rather than low, concluded that the pleural fluid glucose levels were not useful in the differential diagnosis of pleural effusion. In this study, pleural fluid glucose was determined in 46 pleural effusions from various causes to evaluate the usefulness in the differential diagnosis of pleural effusion. In addition, the protein concentration and the electrophoretic patterns of protein and amylases in pleural fluid was compared with that of serum. And the results were as follows. 1. The mean glucose concentration of pleural fluid was 80.8 mg/100 ml in 22 tuberculous origin, 92.5 mg/100 ml in 12 cancer patient and 70.4 mg/100 ml in 10 undiagnosed cases. In 2 cases of paragonimiasis the pleural fliud glucose levels were low (mean, 32.0 mg/100 ml). The percentage of pleural fluid protein to serum is about 75% in all disease groups and the protein level of tuberculous pleural fluid was significantly correlated with that of serum. 2. The disc eletrophoretic patterns of pleural fluid were almost similar with that of serum in all disease groups but the prealbumin fraction was not observed in pleural fluid. 3. With the isoelectric focusing, 4 to 7 isoamylase was observed in serum and the isoelectric point was ranged from pH 5.8 to 7.8 and isoelectic point of main fracticn is pH 7.2. The isoelectic focusing patterns of amylase of pleural fluid were identical to that of serum in all disease group. With the above results it is concluded that the pleural fluid is exudate of serum and that the glucose levels of pleural fluid are not useful in the differential diagnosis of pieural effusions.

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Improvement of Artificial Antibody Secretion Using Supercharged Protein (단백질의 과전하화를 이용한 인공 항체의 분비 개선)

  • Park, Jiyeon;Choi, Heeju;Lee, Hyejin;Ahn, Jung Hoon
    • Journal of Life Science
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    • v.30 no.5
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    • pp.420-427
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    • 2020
  • A repebody, an artificial non-immunoglobulin protein scaffold, is expected to be a solution in the search for faster, cheaper, and customizable antibodies. However, the production of medical repebodies remains difficult due to their low yield and the complex purification processes required. The Pseudomonas fluorescens ABC transporter system has been suggested as an efficient and cost-effective method for repebody production, but the total yield is low because of the secreted protein's positive charge; thus, a repebody with a high isoelectric point needs to be changed into a more negatively charged protein for better secretion. To achieve this, we first attached oligo-aspartic acids to the N- and C-terminals of the repebody, but secretion efficiency was not enhanced significantly. Subsequently, we devised an alternative method for improved secretion efficiency by engineering fifteen positively charged amino acids to aspartic acid in the non-antigen binding sites of the repebody to give a high net negative charge. As a result, secretion efficiency was greatly enhanced from 21.2% (wildtype) to 58.5% (negatively supercharged). The negatively supercharged repebody was succussfully produced extracellularly by ABC transporter secretion system in P. fluorescens.

Effect of Amine Oxide Zwitterionic Surfactant on Characteristics of Liposome (아민 옥사이드 양쪽성 계면활성제 첨가가 리포좀 특성에 미치는 영향에 관한 연구)

  • Mo, DaHee;Lee, SuMin;Lee, JuYeon;Han, DongSung;Lim, JongChoo
    • Applied Chemistry for Engineering
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    • v.27 no.3
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    • pp.291-298
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    • 2016
  • In this study, zwitterionic surfactants were added to liposome systems at different pH conditions to understand the effect of surfactants on liposome characteristics. For this purpose, amine oxide surfactants having different hydrocarbon chain lengths were synthesized and the structure of the resulting product was elucidated by using $^1H$ NMR, $^{13}C$ NMR, and FT-IR. In addition, the physical properties of newly synthesized surfactants such as critical micelle concentration (CMC), surface tension and isoelectric point were measured. The stability characteristics of liposome systems including average particle sizes and zeta potentials were measured by varying pH and hydrocarbon chain lengths of an amine oxide surfactant. Effects of the pH and hydrocarbon chain length of an amine oxide surfactant on fluidity of a liposome membrane were also examined by measuring the deformability and the binding degree between the surfactant and liposome.

Characterization of Thermolabile Pectinesterase and Thermostable Pectinesterase Separated from Valencia Orange (Valencia 오렌지로부터 분리 정제한 비내열성 및 내열성 Pectinesterase의 성질)

  • Hou, Won-Nyoung;M.R., Marshall
    • Korean Journal of Food Science and Technology
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    • v.27 no.5
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    • pp.666-672
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    • 1995
  • This study was carried out to characterize thermolabile pectinesterase (TLPE) and thermostable pectinesterase (TSPE) separated from crude PE of Valencia orange in order to investigate the preventive measures of cloudy juice clarification. The TLPE was observed to be mixture of several isoenzymes with the same molecular weight of 36 KD (37.5 KD) but different isoelectric point of pH 8.4, 8.7, 8.9, 9.8 and ${\geq}10$ which were unstable at $70^{\circ}C$, and the TSPE also was found to be mixture of two or three isoenzymes with the same molecular weight of 53 KD (50 KD) but different isoelectric point of pH 8.7, 9.2 and ${\geq}10$ which had slightly different stability from one another at $70^{\circ}C$. The TLPE and the TSPE had the optimum reaction pH of 7.0 and $7.0{\sim}8.5,\;appK_{M}$ of 1.1 and 1.7 mg/ml, appVmax of 0.53 and $1.01\;{\mu}mol/min/{\mu}g$, and the turnover number of 19.000 and 54,000 mol/mol/min toward Kodak pectin, respectively. The TSPE had higher storage stabiblity and cloud loss effect on orange juice than the TLPE. Above all, the crude PE was most effective on orange juice cloud loss among the PEs used.

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Effects of Mg on corrosion resistance of Al galvanically coupled to Fe (Fe와 galvanic couple된 알루미늄의 내식성에 미치는 마그네슘의 영향)

  • Hyun, Youngmin;Kim, Heesan
    • Corrosion Science and Technology
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    • v.12 no.1
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    • pp.40-49
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    • 2013
  • Effects of magnesium and pH on corrosion of aluminum galvanically coupled to iron have studied by using potentio- dynamic and static tests for polarization curves, Mott-Schottky test for analysis of semiconductor property, and GD-AES and XPS for film analysis. Pitting potential was sensitive to magnesium as an alloying element but not to pH, while passive current was sensitive to pH but not to magnesium. It was explained with, instead of point defect model (PDM), surface charge model describing that the ingression of chloride depends on the state of surface charge and passive film at film/solution interface is affected by pH. In addition, galvanic current of aluminum electrically coupled to iron was not affected by magnesium in pH 8.4, 0.2M citrate solution but was increased by magnesium at the solution of pH 9.1. The galvanic current at pH 9.1 increased with time at the initial stage and after the exposure of about 200 minute, decreased and stabilized. The behavior of the galvanic current was related with the concentration of magnesium at the surface. It agreed with the depletion of magnesium at the oxide surface by using glow discharge atomic emission spectroscopy (GD-AES). In addition, pitting potential of pure aluminum was reduced in neutral pH solution where chloride ion maybe are competitively adsorbed on pure aluminum. It was confirmed by the exponential decrease of pitting potential with log of [$Cl^-$] around 0.025 M of [$Cl^-$] and linear decrease of the pitting potential. From the above results, unlike magnesium, alloying elements with higher electron negativity, lowering isoelectric point (ISE), are recommended to be added to improve pitting corrosion resistance of aluminum and its alloys in neutral solutions as well as their galvanic corrosion resistance in weakly basic solutions.

Studies on the Utilization of Keratins (경단백질(硬蛋白質)의 이용(利用)에 관(關)한 연구(硏究))

  • Suh, Hong-Kyl
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.5 no.1
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    • pp.75-80
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    • 1976
  • The development of protein utilization with keratin(cattle hair and human hair) have heen investigated. The include a) conditioning of hydrolysis of keratin, b) isolation of crude protein, c) pepsin HCl digestibility of isolated protein, d) the amino acid composition of isolated protein and amino acid concentrate. The results are obtained as follow ; 1. Human hair and cattle hair were softened and hydrolysed with hot dilute alkali(especially 1% NaOH) 2. Crude protein were isolated from hydrolysed keratin by means of different solubility and isoelectric point. 3. The protein isolated was of good quality on the point of color, protein content and pepsin digestibility. 4. The amino acid concentrate obtained by acids processing was of good quality on the point of color, taste and amino acid composition.

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