scholarly journals Properties of Peanut (KAC431) Protein Hydrolysates and Their Impact on the Quality of Gluten-Free Rice Bread

Foods ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 942 ◽  
Author(s):  
Suphat Phongthai ◽  
Nuttapon Singsaeng ◽  
Rossarin Nhoo-ied ◽  
Thipubol Suwannatrai ◽  
Regine Schönlechner ◽  
...  
Keyword(s):  
Amino Acids ◽  
Functional Properties ◽  
Positive Impact ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Hydrolysates ◽  
Degree Of Hydrolysis ◽  
Major Protein ◽  
Gluten Free ◽  
Major Protein Band

Protein hydrolysates (PH) with a degree of hydrolysis (DH) of 5%, 10%, and 13% from two varieties of peanut were prepared using two commercial enzymes, Alcalase and Flavourzyme. The content of essential amino acids (30,290 mg/100 g) and hydrophobic amino acids (34,067 mg/100 g) of the peanut variety Kalasin 2 (KAC431) protein was higher than that of a common variety, Kalasin 1 (KAC1) (p < 0.05). The protein molecular weight distributions of the two varieties of peanut detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) were similar, ranging from 15 to 75 kDa, with a major protein band at 50–75 kDa. The antioxidant and functional properties of derived PHs were influenced by DH. Although the foaming ability of protein was improved by DH5%, it was obviously decreased upon increasing DH further. The best emulsifying properties were observed in PH with DH5% (p < 0.05). The incorporation of PH with a small DH, especially when produced using Flavourzyme, had a highly positive impact on the specific volume and relative elasticity of gluten-free bread. The effect of PHs on bread quality was highly correlated with their functional properties. This study suggests that partially enzymatically modified proteins are suitable for incorporation in food products such as bread and other gluten-free products.

Partial purification and characterization of isopenicillin N epimerase activity from Streptomyces clavuligerus

1983 ◽  
Vol 29 (11) ◽  
pp. 1526-1531 ◽  
Author(s):  
Susan E. Jensen ◽  
Donald W. S. Westlake ◽  
Saul Wolfe
Keyword(s):  
Pyridoxal Phosphate ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Streptomyces Clavuligerus ◽  
Enzymic Activity ◽  
Partial Purification ◽  
Major Protein ◽  
Major Protein Band ◽  
Isopenicillin N

Epimerase activity, which converts isopenicillin N to penicillin N, has been partially purified from cell-free extracts of Streptomyces clavuligerus. No stimulating cofactors of this activity were found, and neither EDTA nor anaerobic incubation caused significant inhibition of activity. Although pyridoxal phosphate did not stimulate epimerase activity, the presence of this cofactor was necessary for the stabilization of enzymic activity during the purification process. Epimerase activity was purified 35.5-fold by a combination of salt precipitation, gel filtration, and ion exchange chromatography. Gel filtration indicated that the epimerase has a molecular weight of 60 000 and sodium dodecyl sulphate – polyacrylamide gel electrophoresis of the 35.5-fold purified epimerase showed a major protein band running near that location. Pyridoxal phosphate antagonists did not uniformly inhibit epimerase activity, but the inhibitory effect of hydroxylamine could be partially reversed by pyridoxal phosphate.

The effect of ageing cooled milk on the composition of the fat globule membrane

1972 ◽  
Vol 39 (1) ◽  
pp. 95-105 ◽  
Author(s):  
M. Anderson ◽  
G. C. Cheeseman ◽  
Dorothy J. Knight ◽  
W. F. Shipe
Keyword(s):  
Gel Electrophoresis ◽  
Milk Fat ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Sodium Deoxycholate ◽  
Protein Band ◽  
Major Protein ◽  
Fresh Milk ◽  
Major Protein Band ◽  
Fat Globule

SummaryThe effect of ageing fresh milk for 24 h at 4°C on the composition of 4 fractions of milk fat globule membrane (FGM) – microsomal pellet membrane (MPM), deoxycholate soluble membrane (DOCM), high density membrane (HDM) and low density membrane (LDM) – prepared from washed cream treated with sodium deoxycholate (DOC), was studied for milk of individual cows. Total FGM and its fractions were solubilized by treatment with sodium dodecyl sulphate (SDS), EDTA and β-mercaptoethanol, and the dissociated FGM proteins were separated by polyacrylamide gel electrophoresis in the presence of SDS.Ageing resulted in a greater loss of phospholipid during cream preparation than was found with fresh milk, and also in a reduction in the total amount of FGM isolated. Loss of FGM material was confined to MPM, DOCM and LDM and was accompanied by compositional changes in DOCM and LDM, quantitatively the most significant fractions. Ageing also produced changes in the gel electrophoresis patterns of DOCM, where the band of greatest mobility (mol. wt about 16000) was partially lost, and of LDM, where there was an increase observed in the major protein band.The implication of the results on the proposed models of FGM structure is discussed.

scholarly journals ACE Inhibitory and Antioxidative Bioactive Peptides Derived from Hydrolyzed Soy Milk

Molekul ◽  
2019 ◽  
Vol 14 (1) ◽  
pp. 56
Author(s):  
Sandra Hermanto ◽  
Annisa Septiana ◽  
Deni K Putera ◽  
Fitriah Hatiningsih ◽  
Anna Muawanah
Keyword(s):  
Milk Protein ◽  
Bioactive Peptides ◽  
Antioxidative Activity ◽  
Positive Impact ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Antihypertensive Activity ◽  
Degree Of Hydrolysis ◽  
Soy Milk ◽  
Dpph Method

Soy milk is a soybean processed product rich in protein as well as sources of bioactive peptides. Bioactive peptides defined as specific protein fragments that have a positive impact on body functions and conditions and may ultimately influence health. This study was conducted to explore the potential of hydrolyzed soy milk as a source of antioxidative and antihypertensive bioactive peptides through enzymatic hydrolysis. The initial treatment of soy milk protein was acidic precipitation with hydrochloric acid. Furthermore, protein precipitate was hydrolyzed using pepsin proteolytic enzyme with an enzyme: substrate ratio (1:5, 1:10 and 1:20). Protein hydrolysis was carried out for 0–48 hours at 37 °C in an acetate buffer pH 4.5. The soy milk protein hydrolysates were subjected to determination of % DH (Degree of Hydrolysis) and protein profile by SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis). The antihypertensive assay was carried out by in vitro inhibition of the ACE enzyme (Angiotensin Converting Enzyme) and antioxidative activity assay using the DPPH method. The results showed that the optimum conditions for hydrolysis of soy milk were obtained at 40 hours with a % DH value of 53.24% in enzyme ratio 1:20 and the highest antihypertensive activity was obtained from 48 hours hydrolysis with % ACE inhibition value of 79.31%. The highest antioxidative activity of bioactive peptide was obtained at hydrolysis 48 hours with IC50 69.1034 µg / ml. After fractionated and LCMS characterized it was obtained 2 bioactive peptides with molecular weights of 8.954 and 2,696 kDa. These bioactive peptides from hydrolyzed of soymilk might be potential as an antihypertensive agent and reduce oxidative stress.

scholarly journals β-1,3-Glucanase Activity in Peanut Seed (Arachis hypogaea) is Induced by Inoculation with Aspergillus flavus and Copurifies with a Conglutin-Like Protein

2005 ◽  
Vol 95 (5) ◽  
pp. 506-511 ◽  
Author(s):  
X. Q. Liang ◽  
C. C. Holbrook ◽  
R. E. Lynch ◽  
B. Z. Guo
Keyword(s):  
Arachis Hypogaea ◽  
Aspergillus Flavus ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Major Protein ◽  
Peanut Seed ◽  
Glucanase Activity ◽  
Infected Seed ◽  
Major Protein Band ◽  
Resistant Genotypes

Infection of peanut (Arachis hypogaea) seed by Aspergillus flavus and A. parasiticus is a serious problem that can result in aflatoxin contamination in the seed. Breeding resistant cultivars would be an effective approach to reduce aflatoxin accumulation. The objective of this study was to investigate the expression of the pathogenesis-related (PR) protein β-1,3-glucanase and the isoform patterns in peanut seed inoculated with A. flavus. Peanut genotypes GT-YY9 and GT-YY20 (both resistant to A. flavus infection) and Georgia Green and A100 (both susceptible to A. flavus infection) were used in this study. The activities of β-1,3-glucanase were similar in the uninfected seed of all genotypes, but increased significantly in the resistant genotypes after inoculation in comparison with the susceptible genotypes. An in-gel (native polyacrylamide gel electrophoresis [PAGE]) enzymatic activity assay of β-1,3-glucanase revealed that there were more protein bands corresponding to β-1,3-glucanase isoforms in the infected seed of resistant genotypes than in the infected seed of susceptible genotypes. Both acidic and basic β-1,3-glucanase isoforms were detected in the isoelectric focusing gels. Thin-layer chromatography analysis of the hydrolytic products from the reaction mixtures of the substrate with the total protein extract or individual band of native PAGE revealed the presence of enzymatic hydrolytic oligomer products. The individual bands corresponding to the bands of β-1,3-glucanase isoforms Glu 1 to 5 were separated on the sodium dodecyl sulfate-PAGE, resulting in two bands of 10 and 13 kDa, respectively. The sequences of fragments of the 13-kDa major protein band showed a high degree of homology to conglutin, a storage protein in peanut seed. Conglutin is reported as a peanut allergen, Ara h2. Our data provide the first evidences for peanut having β-1,3-glucanase activities and the association with the resistance to A. flavus colonization in peanut seed. We have not directly demonstrated that conglutin has β-1,3-glucanase activity.

scholarly journals Survey and Characterization of Viruses in Sweetpotato from Zimbabwe

Plant Disease ◽  
1997 ◽  
Vol 81 (10) ◽  
pp. 1115-1122 ◽  
Author(s):  
Farayi Chavi ◽  
A. Ian Robertson ◽  
Benedictus J. M. Verduin
Keyword(s):  
Amino Acids ◽  
Coat Protein ◽  
Sodium Dodecyl ◽  
Protein Band ◽  
Mottle Virus ◽  
Major Protein ◽  
Degenerate Primers ◽  
Vein Clearing ◽  
Major Protein Band

Thirty-one clones of sweetpotatoes collected from some parts of Zimbabwe were used as inoculum sources to mechanically inoculate 13 experimental hosts: Chenopodium amaranticolor, C. quinoa, Cucumis sativus, Datura stramoniumitalic, Gomphrena globosa, Ipomoea purpurea, I. quamoclit, I. rubrocorulea, Nicotiana benthamiana, N. clevelandii, N. glutinosa, N. rustica, and N. tabacum. Systemic vein clearing was observed in N. benthamiana inoculated with buffered sap from nine clones. Purification of the vein clearing inducing agent from one of the sweetpotato clones gave yields ranging from 2 to 17 mg/kg and the A260nm/A280nm was around 1.2. Electron microscopy revealed flexuous filamentous particles with a modal length of 830 nm. Protein analysis of purified virus preparations by sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed a major protein band of 40 kDa, and this was assumed to be the viral coat protein. Minor protein bands of 27, 37, and 46 kDa were also observed. The viral protein degraded upon storage at 4°C over time to yield a protein band of 27 kDa. Polyclonal antiserum was produced against the purified virus. Protein A gold labeling of the purified virus incubated with available antisera; sweetpotato chlorotic stunt virus (SPCSV), sweetpotato feathery mottle virus strain russet crack (SPFMV-RC), sweetpotato feathery mottle virus, sweetpotato mild mottle virus, sweetpotato latent virus, sweetpotato chlorotic fleck virus, and sweetpotato caulimo-like virus resulted in a higher labeling density with the antiserum of SPFMV-RC than with the antiserum of SPCSV, while the other sera did not react. Further characterization of the vein clearing inducing agent was attempted by reverse transcription-polymerase chain reaction amplification of total RNA with degenerate primers for potyviruses and an oligo dT primer and PCR products of correct size were obtained. The nucleotide sequence was determined and the amino acid of the polyprotein deduced. Comparison with other strains of SPFMV showed strong similarity except for an insertion of 22 amino acids at the N-terminus of the coat protein. The coat protein size of 335 amino acids is the biggest SPFMV so far determined.

scholarly journals Purification and properties of pyruvate kinase from the hepatopancreas of Carcinus maenas

1976 ◽  
Vol 153 (1) ◽  
pp. 127-134 ◽  
Author(s):  
I G Giles ◽  
P C Poat ◽  
K A Munday
Keyword(s):  
Pyruvate Kinase ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Carcinus Maenas ◽  
Deae Cellulose ◽  
Protein Band ◽  
Polyacrylamide Gel ◽  
Major Protein ◽  
Major Protein Band

Pyruvatekinase from the hepatopancreas of the common shore crab, Carcinus maenas, was purified to a specific activity of 240 units/mg of protein in the assay conditions described. 2. In one method of purification the enzymic activity could be resolved into two fractions after chromatography on DEAE-cellulose. Fructose 1, 6-diphosphate was able to effect the conversion of one form (peak 1) into the second (peak 2). 3. In the presence of a saturating concentration of fructose 1, 6-diphosphate both forms of the enzyme were kinetically similar. 4. Polyacrylamide-gel electrophoresis of the enzyme 1 day after preparation showed a single protein band. On storage at least three protein bands became visible, all of which were associated with pyruvate kinase activity. 5. Chromatography of the enzyme on Sephadex G-200 indicated a mol.wt. of 247000, but in the presence of fructose 1, 6-diphosphate the elution volume of the enzyme increased corresponding to a mol.wt. of 193000. 6 Dissociation of the enzyme in sodium dodecyl sulphate and 2-mercaptoethanol followed by polyacrylamide-gel electrophoresis produced one major protein band with a mol.wt. of 55000.

Functional Properties of the Protein Isolate and Major Fractions of Pine Nut Proteins Prepared from the Changbai Mountain in China

2014 ◽  
Vol 881-883 ◽  
pp. 766-775 ◽  
Author(s):  
Dan Wu ◽  
Wei Hong Min ◽  
Jing Sheng Liu ◽  
Li Fang ◽  
Hong Mei Li ◽  
...  
Keyword(s):  
Amino Acids ◽  
Functional Properties ◽  
Protein Solubility ◽  
Essential Amino Acids ◽  
Protein Isolate ◽  
Absorption Capacity ◽  
Changbai Mountain ◽  
Major Protein ◽  
Pine Nut ◽  
A Value

The functional properties of protein isolate and major protein fractions prepared from Changbai Mountain pine nuts were investigated. Albumin, globulin, glutelin, and protein isolates were obtained after the Osborne method and alkaline dissolution and acid precipitation, and protein contents of the fractions are 48.02%, 81.93%, 83.02%, and 89.69%, respectively. For the sulfhydryl contents, albumin is the highest, and glutelin is the lowest. In a disulphide bond, the protein isolate content is the highest with a value of 28.74 μmol/g, and the glutelin content is the lowest with the value of 13.46 μmol/g. For the four kinds of proteins, the essential amino acids in percentage of total amino acids are 31.13%, 34.22%, 30.30%, and 34.54%, respectively. The pH dependent protein solubility profile reveals that the minimum solubility is at pH 5.0, which corresponds to the isoelectric point. Protein isolate has the minimum water absorption capacity with a value of 0.59 ml/g. On the other hand, albumin has the minimum oil absorption capacity with a value of 2.11 ml/g. The emulsifying activity and stability and the foaming activity and stability increased with increasing concentration of four kinds of proteins. SDS-PAGE results showed that these four kinds of proteins have different molecules.

A metalloproteinase extracellularly released byCrithidia deanei

2003 ◽  
Vol 49 (10) ◽  
pp. 625-632 ◽  
Author(s):  
Claudia Masini d'Avila-Levy ◽  
Rodrigo F Souza ◽  
Rosana C Gomes ◽  
Alane B Vermelho ◽  
Marta H Branquinha
Keyword(s):  
Molecular Mass ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Residual Activity ◽  
Major Protein ◽  
Motile Cells ◽  
Sds Page ◽  
Triton X

Actively motile cells from a cured strain of Crithidia deanei released proteins in phosphate buffer (pH 7.4). The molecular mass of the released polypeptides, which included some proteinases, ranged from 19 to 116 kDa. One of the major protein bands was purified to homogeneity by a combination of anion-exchange and gel filtration chromatographs. The apparent molecular mass of this protein was estimated to be 62 kDa by sodium dodecyl sulfate – polyacrylamide gel electrophoresis (SDS–PAGE). The incorporation of gelatin into SDS–PAGE showed that the purified protein presented proteolytic activity in a position corresponding to a molecular mass of 60 kDa. The enzyme was optimally active at 37 °C and pH 6.0 and showed 25% of residual activity at 28 °C for 30 min. The proteinase was inhibited by 1,10-phenanthroline and EDTA, showing that it belonged to the metalloproteinase class. A polyclonal antibody to the leishmanial gp63 reacted strongly with the released C. deanei protease. After Triton X-114 extraction, an enzyme similar to the purified metalloproteinase was detected in aqueous and detergent-rich phases. The detection of an extracellular metalloproteinase produced by C. deanei and some other Crithidia species suggests a potential role of this released enzyme in substrate degradation that may be relevant to the survival of trypanosomatids in the host.Key words: endosymbiont, trypanosomatid, extracellular, proteinase.

scholarly journals Enzymatic hydrolysis of defatted soy flour by three different proteases and their effect on the functional properties of resulting protein hydrolysates

2011 ◽  
Vol 20 (No. 1) ◽  
pp. 7-14 ◽  
Author(s):  
M. Hrčková ◽  
M. Rusňáková ◽  
J. Zemanovič
Keyword(s):  
Enzymatic Hydrolysis ◽  
Functional Properties ◽  
Aqueous Dispersion ◽  
Protein Hydrolysates ◽  
Soy Flour ◽  
Degree Of Hydrolysis ◽  
Defatted Soy Flour ◽  
Sds Page ◽  
Soy Protein Hydrolysates ◽  
Hydrolysis Of

Commercial defatted soy flour (DSF) was dispersed in distilled water at pH 7 to prepare 5% aqueous dispersion. Soy protein hydrolysates (SPH) were obtained by enzymatic hydrolysis of the DSF using three different proteases (Flavourzyme 1000 L, No-vozym FM 2.0 L and Alcalase 2.4 L FG). The highest degree of hydrolysis (DH 39.5) was observed in the presence of protease Flavourzyme. SPH were used for measuring functional properties (foaming stability, gelation). Treatment with Flavourzyme improved foaming of proteins of DSF. Foaming stability was low in the presence of Novozym. Proteases treated DSF showed good gelation properties, mainly in the case of treatment with Flavourzyme. SDS-PAGE analysis showed that after enzyme ad-dition to the 5% aqueous dispersion of DSF each enzyme degraded both b-conglycinin and glycinin. In general, the basic polypeptide from glycinin showed the highest resistance to proteolytic activity. The most abundant free amino acids in the hydrolysates were histidine (30%), leucine (24%) and tyrosine (19%) in the case of the treatment with proteases Alcalase and Novozym, and arginine (22.1%), leucine (10.6%) and phenylalanine (12.9%) in the case of the treatment with Flavourzyme. &nbsp;

scholarly journals Simultaneous Ultrasound and Heat Enhance Functional Properties of Glycosylated Lactoferrin

Molecules ◽  
2020 ◽  
Vol 25 (23) ◽  
pp. 5774
Author(s):  
Zhipeng Li ◽  
Dexue Ma ◽  
Yiyang He ◽  
Siqi Guo ◽  
Fuguo Liu ◽  
...  
Keyword(s):  
Functional Properties ◽  
Maillard Reaction ◽  
Spatial Configuration ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Surface Hydrophobicity ◽  
Orthogonal Experimental Design ◽  
Maillard Reaction Products ◽  
Reaction Products ◽  
Ultrasound Assisted

Protein-polysaccharide covalent complexes exhibit better physicochemical and functional properties than single protein or polysaccharide. To promote the formation of the covalent complex from lactoferrin (LF) and beet pectin (BP), we enhanced the Maillard reaction between LF and BP by using an ultrasound-assisted treatment and studied the structure and functional properties of the resulting product. The reaction conditions were optimized by an orthogonal experimental design, and the highest grafting degree of 55.36% was obtained by ultrasonic treatment at 300 W for 20 min and at LF concentration of 20 g/L and BP concentration of 9 g/L. The formation of LF-BP conjugates was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Fourier transform infrared (FTIR) spectroscopy. Ultrasound-assisted treatment can increase the surface hydrophobicity, browning index, 1,1-diphenyl-2-picryl-hydrazyl (DPPH) and 2,2’-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) free radicals scavenging activity of LF due to the changes in the spatial configuration and formation of Maillard reaction products. The thermal stability, antioxidant activity and emulsifying property of LF were significantly improved after combining with BP. These findings reveal the potential application of modified proteins by ultrasonic and heat treatment.

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