Comparison of pectolytic and proteolytic activities of six clarifying commercial preparations. Incidence of secondary enzymatic activities on proteins in Champagne musts

<p style="text-align: justify;">In order ta estimate c1arifying aptitude and the effects of prefermentation treatment by commercial pectinases, polygalacturonase (PG), pectin Iyase (PL), pectinmethylesterase (PME) and proteolytic activities of six pectinases samples were studied. PG activity is preponderate and proportional to the relative protein concentration.</p><p style="text-align: justify;">These one, submitted to both SDS-PAGE and isoelectric focusing technics, show molecular weights ranging from 20 ta 100 kDa and isoelectric points ranging from 3.00 ta 7.40. Total protein of Champagne grape varieties Chardonnay (CH) and Pinot noir (PN) - were submitted to an isoelectric focusing. All of the pI's ranging from 3.30 to 4.55 (CH) and from 3.30 to 4.90 (PN). Proteases are put in evidence in commercial pectinases using fluorescamine- casein as substrate. However, they seem to have no incidence upon must proteins.</p>

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Quality Attributes of Ultra-High Temperature-Treated Model Beverages Prepared with Faba Bean Protein Concentrates

Foods ◽

10.3390/foods10061244 ◽

2021 ◽

Vol 10 (6) ◽

pp. 1244

Author(s):

Malik Adil Nawaz ◽

Tanoj Kumar Singh ◽

Regine Stockmann ◽

Hema Jegasothy ◽

Roman Buckow

Keyword(s):

Physicochemical Properties ◽

Faba Bean ◽

Protein Concentration ◽

High Concentration ◽

Molecular Weights ◽

Oil In Water ◽

Sds Page ◽

Lower Protein ◽

7S Globulin ◽

Bean Protein

The objective of this research was to develop a model faba bean drink with a high concentration of protein (>4% w/w). The protein molecular weights and frequency for both faba and soy were assessed using SDS-PAGE. Results showed similarities in the protein molecular weight of both faba and soy (mainly 11S globulin ~Glycinin and 7S globulin ~β-conglycinin). Thus, faba can be considered as a potential soy replica in plant-based milk beverages. Oil-in-water emulsions (5–8% w/w available protein) were prepared using faba bean protein concentrate (FPC), 1% sunflower oil, and 0.2% sunflower lecithin. These emulsions were used as model beverages and were further investigated for UHT processibility, stability, and physicochemical properties. The physicochemical properties of emulsions at various processing stages viz., coarse emulsification, homogenisation, and UHT, were measured. An increase in the protein concentration and thermal treatment resulted in an increased oil droplet size, coalescence and flocculation, and protein aggregation. Lower protein concentrations viz., 5–6%, showed greater negative ζ-potential, and thereby, high dispersibility through enhanced electrostatic repulsions than those of higher concentrations (7–8%). Furthermore, an increase in protein concentration and UHT treatment resulted in an increased creaming index. In total, 21 different volatile compounds were detected and quantified, representing different chemical classes, namely alcohols, aldehydes, ketones, esters, furan, and acids. These volatiles have major consequences for the overall flavour chemistry of the model beverage product. Overall, this study showed the potential for application of faba bean as a protein source in UHT-treated legume-based beverages and identified areas for further development.

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Erwinia amylovora Enfeksiyonu Sonrası Elma, Armut ve Ayva Çeşitlerinde Konukçu Protein Miktarlarının Belirlenmesi

Turkish Journal of Agriculture - Food Science and Technology ◽

10.24925/turjaf.v3i3.154-163.246 ◽

2014 ◽

Vol 3 (3) ◽

pp. 154

Author(s):

Şerife Çetin ◽

Kubilay Kurtuluş Baştaş

Keyword(s):

Protein Content ◽

Total Protein ◽

Erwinia Amylovora ◽

Bacterial Pathogen ◽

Molecular Weights ◽

Molecular Tests ◽

Sds Page ◽

Apple Cultivars ◽

Blight Disease ◽

Santa Maria

Fire blight disease caused by Erwinia amylovora is a destructive bacterial pathogen mainly on pears, apples and quinces from Rosaceae family. In this study, it was aimed determination of total protein amounts in different apple cultivars (Braeburn, Fuji, Gala and Golden), pear cultivars (Santa Maria and Williams) and quince cultivars (Eşme and Ekmek) in the infections of two virulent E. amylovora strains (Ea234-1 and Ea240-3) according as the time. It was taken leaf samples after leaf inoculation with E. amylovora (108 CFU ml-1) at 24th, 36th and 72nd hours. For verification of the infections, re-isolations were made from bacteria inoculated plants and the agent was identified as E. amylovora by biochemical, physiological and molecular tests. In determining the amounts of total protein and in the SDS-PAGE analyses were used Bradford and Laemmli methods, respectively, and absorbance values of protein extracts derived from the leaf samples taken, were obtained at 595 nm wavelength. According to the findings obtained; after infection of E. amylovora in the apple varieties comparing to controls, total protein concentrations at 24th hours increased and a decrease in the amount of 36th to 72nd hours and Braeburn has the highest protein content was determined. In the pear varieties, while total protein concentrations at 24th and 36th hours increased, a decrease in the amount of 72nd hour, and Santa Maria variety has the highest protein content was detected. In the quince varieties, total protein concentrations at 72th hour increased and Eşme variety has the highest protein content was identified. As a result of SDS-PAGE analysis, protein fractions which have different molecular weights were obtained. The protein bands were defined approximately 55-70 kDa and 35-55 kDa molecule weight on apple and quince varieties, respectively and also approx. 55-70 kDa in pear varieties.

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Affinity Chromatography of Human Factor VIII Using Human and Rabbit Antibodies to Factor VIII

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657026 ◽

1979 ◽

Vol 42 (04) ◽

pp. 1306-1315 ◽

Cited By ~ 3

Author(s):

Janet L Lane ◽

H Ekert ◽

A Vafiadis

Keyword(s):

Molecular Weight ◽

Factor Viii ◽

Affinity Chromatography ◽

Protein Concentration ◽

Gel Filtration ◽

Subunit Structure ◽

Molecular Weights ◽

Human Factor Viii ◽

Sds Page ◽

Normal Human

SummaryFactor VIII, purified by gel filtration on Sepharose 2B, has an 8 band multiple subunit structure, with molecular weights ranging from 30,000 to 230,000, on reduction and SDS-PAGE at a protein concentration of 400 μg/gel. Affinity chromatography of this factor VIII preparation with insolubilized haemophilic antibody to factor VIII showed that 45-81% VIII:C and 0-33% VIILRag were attached to the column. Elution of the column with 0.25 M CaCl2 did not show VIII:C or VIILRag in the eluate. NH4SCN dissociation of the column, followed by reduction and SDS-PAGE of the dissociated protein, showed that 95 % of the protein bound by haemophilic antibody had a molecular weight similar to the low molecular weight subunits of the reduced factor VIII.In control experiments with normal Human IgG, 3% of VIII:C and 5% of VIILRag were attached to the column. NH4SCN dissociation of the column, followed by reduction and SDS-PAGE of the protein, showed 2 faint bands with molecular weight consistent with heavy and light chains of IgG.Similar experiments with antibody to factor VIII showed that 67-83% of VIILC and 61-76% of VIII:Rag were attached to the column. Elution of the column with 0.25 M CaCl2 showed 10% of the applied VIII:C, but no VIII:Rag in the eluate. NH4SCN dissociation of the column, followed by reduction and SDS-PAGE of the dissociated protein, showed an 8 band subunit structure similar to the reduced factor VIII.

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An electrophoretic characterization of iron-transporting proteins in Mannheimia haemolytica A1

Polish Journal of Veterinary Sciences ◽

10.2478/pjvs-2013-0073 ◽

2013 ◽

Vol 16 (3) ◽

pp. 527-532 ◽

Cited By ~ 1

Author(s):

A. Puchalski ◽

R. Urban-Chmiel ◽

M. Dec ◽

A. Wernicki

Keyword(s):

Immune Response ◽

Iron Uptake ◽

Outer Membrane Proteins ◽

Mannheimia Haemolytica ◽

Iron Ions ◽

Growth Environment ◽

Molecular Weights ◽

Sds Page ◽

Isoelectric Points

AbstractIron-regulated outer membrane proteins (IROMPs) in Mannheimia haemolytica A1, which function as a receptor for complexes containing iron ions, are induced by iron deficiency in the growth environment of the bacteria. Densitometric analysis of SDS-PAGE separation showed expression of IROMPs of 71, 77, and 100 kDa in the case of bacteria grown in a medium with 2,2-dipyridyl. The electrophoregrams obtained in 2-DE separations confirmed the presence of protein fractions with these molecular weights and isoelectric points ranging from 5.4 to 6.4. The results of the study also confirmed the ability of M. haemolytica A1 proteins involved in iron uptake to induce a protective immune response. In Western blot with serum from convalescent calves naturally infected with M. haemolytica A1, distinct reactions were obtained for IROMPs of 71, 77, and 100 kDa.

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External morphological stages and protein variations along with the embryonic development of the longarm river prawn Macrobrachium tenellum (Smith, 1871)

Latin American Journal of Aquatic Research ◽

10.3856/vol49-issue2-fulltext-2605 ◽

2021 ◽

Vol 49 (2) ◽

pp. 202-211

Author(s):

Marcelo U. García-Guerrero ◽

Dulce M. Mateos-Guerrero ◽

Juan J. Alpuche-Osorno ◽

Rodolfo B. De los Santos-Romero

Keyword(s):

Embryonic Development ◽

Protein Concentration ◽

Egg Size ◽

Morphological Changes ◽

Larval Rearing ◽

Polyacrylamide Gels ◽

Protein Extract ◽

Molecular Weights ◽

Sds Page ◽

Page Electrophoresis

A good understanding of a given species' embryology is important to settle the larval rearing bases when juveniles are required for culture purposes or conservation programs. Changes in embryonic morphology, protein concentration, and protein type occurring in prawn eggs were analyzed in the present work. Berried females of Macrobrachium tenellum were collected in the Colotepec River, Oaxaca, Mexico. The eggs were taken from the ovigerous mass and embryonic stages classified by their color. Morphological changes in the embryos allowed identifying six embryonic stages based on color, egg size, and morphological features. Determinations of the protein extract were executed in SDS-PAGE (electrophoresis in polyacrylamide gels) and, subsequently, proteomic analyses were also performed. Protein bands along embryonic development and their molecular weights are presented and commented.

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The purification and properties of the l-serine O-sulphate-degrading system of pig liver

Biochemical Journal ◽

10.1042/bj1210747 ◽

1971 ◽

Vol 121 (5) ◽

pp. 747-752 ◽

Cited By ~ 6

Author(s):

N. Tudball ◽

P. Thomas ◽

R. Bailey-Wood

Keyword(s):

Amino Acid ◽

Isoelectric Focusing ◽

Enzyme System ◽

Gel Filtration ◽

Pig Liver ◽

Molecular Weights ◽

Purification And Properties ◽

Isoelectric Points ◽

Degrading System ◽

Similar Amino Acid

1. The enzyme system from pig liver responsible for the αβ-elimination of l-serine O-sulphate was purified 1000-fold. 2. Isoelectric focusing produced two enzymically active fractions with isoelectric points at pH5.6 and 5.9 respectively. 3. Osmometry and gel filtration showed both enzymes to possess molecular weights of approx. 54000. 4. The separate activities exhibited similar amino acid compositions.

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Isoelectric focusing of insulin, 125I-insulin and the 125I-insulin-antibody complex

Collection of Czechoslovak Chemical Communications ◽

10.1135/cccc19791828 ◽

1979 ◽

Vol 44 (6) ◽

pp. 1828-1834

Author(s):

Asja Šiševa ◽

Jiřina Slaninová ◽

Tomislav Barth ◽

Stephan P. Ditzov ◽

Luben M. Sirakov

Keyword(s):

Isoelectric Focusing ◽

Polyacrylamide Gel ◽

The Other ◽

Insulin Antibody ◽

Isoelectric Points ◽

Antibody Complex ◽

And Storage ◽

Acid Region ◽

Three Components

Isoelectric focusing on polyacrylamide gel columns of three native crystalline commercial preparations of insulin and 125I-labelled insulin was carried out. All the compounds studied contained three components of different isoelectric points. The largest fraction, having pI 5.60 ± 0.05, was common to all preparations. The other two fractions were situated in the acid region of pH between pI 4.5 and 5.2. The presence of these fractions is explained by the contamination of crystalline insulins by proinsulin and by the formation of des-amido derivatives during the dissolving and storage of insulin samples, and, in case of labelled insulin, also by the presence of heavily iodinated insulin and contaminating components. The isoelectric focusing of the complex 125I-insulin-antibody showed a peak of radioactivity having pI 6.15 ± 0.05.

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Isoelectric focusing of glutathione S-transferases from rat liver and kidney

Biochemical Journal ◽

10.1042/bj1750937 ◽

1978 ◽

Vol 175 (3) ◽

pp. 937-943 ◽

Cited By ~ 28

Author(s):

Barbara F. Hales ◽

Valerie Jaeger ◽

Allen H. Neims

Keyword(s):

Substrate Specificity ◽

Isoelectric Focusing ◽

Transferase Activity ◽

Sprague Dawley ◽

Substrate Specificities ◽

Liver Cytosol ◽

Sprague Dawley Rats ◽

Isoelectric Points ◽

Liver And Kidney ◽

Glutathione S Transferases

The glutathione S-transferases that were purified to homogeneity from liver cytosol have overlapping but distinct substrate specificities and different isoelectric points. This report explores the possibility of using preparative electrofocusing to compare the composition of the transferases in liver and kidney cytosol. Hepatic cytosol from adult male Sprague–Dawley rats was resolved by isoelectric focusing on Sephadex columns into five peaks of transferase activity, each with characteristic substrate specificity. The first four peaks of transferase activity (in order of decreasing basicity) are identified as transferases AA, B, A and C respectively, on the basis of substrate specificity, but the fifth peak (pI6.6) does not correspond to a previously described transferase. Isoelectric focusing of renal cytosol resolves only three major peaks of transferase activity, each with narrow substrate specificity. In the kidney, peak 1 (pI9.0) has most of the activity toward 1-chloro-2,4-dinitrobenzene, peak 2 (pI8.5) toward p-nitrobenzyl chloride, and peak 3 (pI7.0) toward trans-4-phenylbut-3-en-2-one. Renal transferase peak 1 (pI9.0) appears to correspond to transferase B on the basis of pI, substrate specificity and antigenicity. Kidney transferase peaks 2 (pI8.5) and 3 (pI7.0) do not correspond to previously described glutathione S-transferases, although kidney transferase peak 3 is similar to the transferase peak 5 from focused hepatic cytosol. Transferases A and C were not found in kidney cytosol, and transferase AA was detected in only one out of six replicates. Thus it is important to recognize the contribution of individual transferases to total transferase activity in that each transferase may be regulated independently.

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Morph-Specific Proteins in Pollen and Styles of Distylous Turnera (Turneraceae)

Genetics ◽

10.1093/genetics/146.2.669 ◽

1997 ◽

Vol 146 (2) ◽

pp. 669-679

Author(s):

Andreas Athanasiou ◽

Joel S Shore

Keyword(s):

Isoelectric Focusing ◽

Isozyme Markers ◽

Unlinked Locus ◽

Full Expression ◽

Isoelectric Points ◽

Isozyme Loci ◽

Specific Proteins ◽

S Allele

We used nondenaturing isoelectric focusing (IEF) in a survey of plants from 11 populations to identify style and pollen proteins unique to the short-styled morph of Turnera scabra, T. subulata and T. krapovickasii. Three protein bands [approximately isoelectric points (pIs) 6.1, 6.3 and 6.5] were found only in styles and stigmas of short-styled plants while two bands (approximately pIs 6.7 and 6.8, M r 56 and 59 kD) occur only in pollen of short-styled plants. Some of these bands appear very late in development, within 24 hr before flowering. Two isozyme loci were mapped to an 8.7 cM region spanning the distyly locus. Using these isozyme markers we identified progeny exhibiting recombination adjacent to the distyly locus. No recombinants between the distyly locus and the locus or loci controlling the presence of the short-styled morph-specific proteins were obtained. This suggests that the loci encoding these proteins are either extremely tightly linked to the distyly locus and in complete disequilibrium with the S allele or exhibit morph-limited expression. Crosses to a plant showing an unusual style protein phenotype demonstrated that an additional unlinked locus is required for full expression of the style proteins. The function of the morph-specific proteins is unknown

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