scholarly journals Effects of extraction methods on protein properties obtained from paddy rice and germinated paddy rice

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11365
Author(s):  
Wirot Likittrakulwong ◽  
Pisit Poolprasert ◽  
Khongsak Srikaeo
Keyword(s):  
Phase Transition ◽  
Amino Acids ◽  
Antioxidant Capacity ◽  
Sodium Dodecyl ◽  
Extraction Methods ◽  
Paddy Rice ◽  
Protein Fractions ◽  
Major Protein ◽  
Enzymatic Extraction ◽  
Sds Page

Rice protein has attracted considerable attention recently due to its physiological effects. This study extracted the proteins from paddy rice (PR) and germinated paddy rice (GPR) using three methods i.e., alkaline, sodium dodecyl sulfate (SDS) reagent and enzymatic extractions. The extracted proteins or protein fractions were assessed for their properties using various techniques. Data were analyzed by 2′3 factorial design experiment. It was found that germination and extraction methods significantly affected the concentration of protein fractions when analyzed by Bradford assay. Average protein fraction concentration of the GPR was lower than that of PR. SDS-PAGE patterns of protein fractions obtained from PR and GPR using any extraction method displayed similar protein profiles. Three major protein bands at about 13 kDa (prolamin), 22–23 kDa (basic glutelin) and 37–39 kDa (acidic glutelin) with small amount of 57 kDa proglutelin were observed. For amino acid profile, germination increased the content of most amino acids, resulting in the higher content of amino acids in GPR, excepted for some amino acids. When processed with in vitro digestion, protein fractions from GPR exhibited a higher level of digestibility than those from PR as evidenced by the less intensity of the protein bands obtained from SDS-PAGE. Alkaline and SDS reagent extractions provided more digestible protein fractions than enzymatic extraction. Extraction methods also influenced phase transition of protein fractions as investigated by a DSC. Alkaline extraction resulted in protein fractions with higher phase transition temperature than the other methods. For antioxidant capacity, extraction methods as well as germination significantly affected antioxidant capacity of the protein fractions. Enzymatic extraction provided protein fractions with the best antioxidant capacity.

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 Evaluation of effective protein extraction procedure to profile petiole of Moringa oleifera

2020 ◽  
Vol 7 (2) ◽  
pp. 214
Author(s):  
Zetty Amirah Zulkifli ◽  
Zaidah Rahmat
Keyword(s):  
Moringa Oleifera ◽  
Protein Extraction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Extraction Procedure ◽  
Electrophoretic Pattern ◽  
Antioxidant Property ◽  
Extraction Methods ◽  
Protein Profiling ◽  
Sds Page

Moringa oleifera is widely known as multipurpose tree since all of its parts confer multiple functions. The leaf is highly favourable among consumers while the petiole is mostly wasted. There are numerous studies on the flavonoid and antioxidant property of the stem and twig. However, study on the petiole has never been done. There-upon, this study was conducted to develop protein profiling of the petiole. In this study, 6 different protein extraction methods were tested on the fresh petiole before its protein quantity and quality were checked via Bradford assay and Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) respectively. The in-solution digestion was then done prior to LC-MS/MS analysis. The protein electrophoretic pattern from the SDS-PAGE proves that method 6 using Tris HCl buffer with incorporation of dithiothreitol (DTT) and phenylmethylsulfonyl fluoride (PMSF) confers the best quality of protein. It produced the highest number of visible individual bands compared to other methods. Meanwhile, 93 proteins were successfully identified via LCMS analysis where the protein, signal response and carbohydrate metabolism categories confer the highest percentage. High quality and content of the protein extracted from the petiole including the antioxidant, anticancer and antidiabetic protein identified suggested that consuming this part of the plant could enhance nutrients of human body.

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.

scholarly journals Plasma protein concentrations of the young and adult Amazona brasiliensis parrots

2017 ◽  
Vol 71 (1) ◽  
pp. 44-51
Author(s):  
Elizabeth Schmidt ◽  
Patricia Serafini ◽  
Elenise Sipinski ◽  
Antonio Paulillo
Keyword(s):  
Plasma Protein ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Clinical Pathology ◽  
Specific Protein ◽  
Major Protein ◽  
Plasma Protein Concentration ◽  
Protein Gel ◽  
Sds Page

Introduction. The Red-tailed Amazon parrot (Amazona brasiliensis) is an endangered species of the Psittacine family, and for which various data are important for a comprehensive preservation plan. Data about plasma protein gel electrophoresis of Amazon parrot blood are scarce. The purpose of this study was to determine plasma protein concentrations and concentrations of major protein bands in blood of young and adult Red-tailed Amazon parrot (Amazona brasiliensis). Materials and Methods. Blood samples from eight young and eight adult healthy free-living parrots were obtained. Plasma protein concentration and fractions were determined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Mann-Whitney U test was used to compare variables. Results and Conclusions. Six major protein bands with the following molecular weights were identified by SDS-PAGE: 170 kDa, 117 kDa, 85 kDa (putative ovotransferrin), 60 kDa, 45 kDa and 23 kDa. Adult parrots had significantly higher concentrations of total proteins, albumin and other proteins with similar mobility (around 60 kDa). Young birds had significantly higher levels of 23kDa proteins. The concentration of putative ovotransferrin (85 kDa) was not different between young and adult parrots. Plasma protein gel electrophoresis patterns in Red-tailed Amazon parrots are similar between young and adult animals, but specific protein bands differ in their absolute concentrations. This finding should be taken into consideration when clinical pathology data are analysed.

scholarly journals First Report of the Cymbidium Mosaic Potexvirus (CymMV) Infecting the Terrestrial Orchid Phaius tankervilliae in Costa Rica

Plant Disease ◽  
1998 ◽  
Vol 82 (10) ◽  
pp. 1171-1171 ◽  
Author(s):  
L. Moreira ◽  
W. Villalobos ◽  
H. T. Hsu ◽  
E. Rodríguez-Cerezo ◽  
C. Rivera
Keyword(s):  
Costa Rica ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Central Valley ◽  
Major Protein ◽  
Terrestrial Orchid ◽  
First Report ◽  
Foliar Symptoms ◽  
Viral Particles ◽  
Sds Page

In 1996, plants of the terrestrial orchid Phaius tankervilliae from a nursery in the Central Valley of Costa Rica were observed with mild to severe foliar symptoms of chlorotic streak. No differences were observed in growth, bulb production, flowers, or flowering time between symptomatic and asymptomatic plants, except the symptomatic plants had earlier senescence. Occasionally, the flowers displayed symptoms of chlorosis and white rings in the sepals. Extracts from symptomatic leaves were concentrated by differential centrifugation and analyzed after sucrose gradients. Negative staining of fractions from gradients from symptomatic plants showed the presence of filamentous viral particles 500 by 17 nm. Purified particles contained a single major protein of about 28 kDa as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and a single RNA of about 7 kb, which is greater than the 6.2 kb reported (GenBank). These data suggest the presence of a potexvirus in symptomatic plants (1,2). In enzyme-linked immunosorbent assays, symptomatic plants reacted strongly with antiserum specific for Cymbidium mosaic potexvirus (CymMV). This is the first report of CymMV in P. tankervilliae in Costa Rica. References: (1) J. A. Frowd and J. H. Tremaine. Phytopathology 67:43, 1977. (2) H. T. Hsu et al. Phytopathology 82:491, 1992.

scholarly journals Storage protein profile and amino acid content in wild rice Oryza glumaepatula

2013 ◽  
Vol 48 (1) ◽  
pp. 66-72 ◽  
Author(s):  
Karina Freire D'eça Nogueira Santos ◽  
Ricardo Diógenes Dias Silveira ◽  
Claudia Cristina Garcia Martin-Didonet ◽  
Claudio Brondani
Keyword(s):  
Amino Acids ◽  
Wild Species ◽  
Total Protein ◽  
Storage Protein ◽  
Protein Profile ◽  
Protein Fractions ◽  
Interspecific Crosses ◽  
Major Protein ◽  
Rice Cultivars ◽  
Oryza Glumaepatula

- The objective of this work was to determine the total protein profile and the contents of the four major protein fractions (albumin, globulin, prolamin and glutelin) and of the amino acids in the endosperm of the rice wild species Oryza glumaepatula. The experiment was performed with 29 accessions of this species, collected from 13 Brazilian locations, and two commercial cultivars. Protein samples were prepared using dried, polished, and ground grains to obtain homogeneous, dry flour used in the preparation of extracts. Oryza glumaepatula accessions were identified with the highest levels of total protein, albumin and glutelin protein fractions, and amino acids (with the exception of tryptophan) in comparison to the two analized rice cultivars. The albumin and glutelin profiles in SDS-Page were distinct between rice cultivars and O. glumaepatula. This wild species has the potential to increase the nutritional quality of rice storage protein through interspecific crosses.

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.

Pheromaxein, the pheromonal steroid-binding protein, is a major protein synthesized in porcine submaxillary salivary glands

1991 ◽  
Vol 128 (2) ◽  
pp. 205-212 ◽  
Author(s):  
W. D. Booth ◽  
K. I. von Glos
Keyword(s):  
Total Protein ◽  
Binding Protein ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Submaxillary Gland ◽  
Major Protein ◽  
Sds Page ◽  
Gland Tissue ◽  
Steroid Binding ◽  
Steroid Binding Protein

ABSTRACT Submaxillary salivary gland tissue from large White, Göttingen miniature and Meishan (Chinese) breeds of pig, and European wild boars, was incubated with [35S]methionine. The radiolabelled amino acid was incorporated into protein in all incubations as demonstrated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Specifically [35S]methionine was predominantly incorporated into the α- and β-charge isomers of pheromaxein, a 16-androstene steroid-binding protein, as shown by SDS-PAGE in combination with vertical isoelectric focusing on polyacrylamide slab gels. The synthesis of pheromaxein occurred in submaxillary gland tissue from both sexes, including tissues stored frozen at −70 °C for long periods. There was little evidence for pheromaxein synthesis in parotid gland tissue or skeletal muscle. Total protein, pheromaxein and total 16-androstenes were determined in the submaxillary gland cytosols of six mature Göttingen miniature boars and a positive correlation was found between these glandular constituents. The amounts of endogenous pheromaxein relative to total protein in the submaxillary gland cytosols (range 10·3–18·0%), together with the predominant synthesis of this protein in vitro, indicate that pheromaxein is a major protein produced in porcine submaxillary glands, particularly in those of the male. Journal of Endocrinology (1991) 128, 205–212

Protein Distribution in Human Endothelial Cells in Primary Culture Studied by Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophores

1979 ◽  
Author(s):  
T. Børsum ◽  
I. Hagen ◽  
T. Henriksen ◽  
B. Carlander
Keyword(s):  
Endothelial Cells ◽  
Protein Pattern ◽  
Sodium Dodecyl ◽  
Protein Composition ◽  
Major Protein ◽  
Protein Distribution ◽  
Human Endothelial Cells ◽  
Neuraminidase Treatment ◽  
Sds Page ◽  
Pas Staining

The surface structure and protein composition of cultured human endothelial cells were examined by lactoperoxidase catalyzed iodonation followed by SDS-PAGE of the cells. Five major protein bands (MW 224.000, 187.000, 70.000, 52.000 and 42.000) and several minor bands were seen. No differences in the protein pattern were observed between samples from confluent and nonconfluent cultures. In contrast, PAS staining of the gels revealed no glycoprotein bands unless the cells had grown to confluency. Then PAS staining revealed three bands of MW 240.000, 208.000 and 145.000. The fire one may represent fibronectin. The radioavtive iodine was associated with proteins in the 230.000, 145.000, 70.000, 55.000 and 45.000 MW regions. The first two regions showed iodination only in samples from confluent cultures. 33% of the acid-hydrolyzable sialic acid was liberated after neuraminidase treatment of the cells. No difference in the protein or glycoprotein pattern was seen after SDS-PAGE of neuraminidase-treated cells. In conclusion, four of the protein bands were susceptible to iodination, indicating an external localization in the plasma membrane. Two of these were iodinated only when cells had grown to confluency.

scholarly journals Myeloperoxidase: its structure and expression during myeloid differentiation

Blood ◽  
1985 ◽  
Vol 65 (2) ◽  
pp. 484-491 ◽  
Author(s):  
HP Koeffler ◽  
J Ranyard ◽  
M Pertcheck
Keyword(s):  
Relative Rate ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Major Protein ◽  
Trichloracetic Acid ◽  
Major Band ◽  
Minor Band ◽  
Sds Page ◽  
Human Granulocytes ◽  
Normal Human

Abstract Myeloperoxidase (MPO) is a major protein present in myeloid cells and is used by these cells to help kill microbes. The human promyelocytic HL-60 line can be induced to differentiate to granulocytes or macrophagelike cells. Poly (A) containing RNA was isolated from HL-60 granulocytes, HL-60 macrophages, HL-60 blasts, and normal human granulocytes. The mRNA was translated in a reticulocyte lysate system in the presence of 35S-methionine. The MPO was precipitated from the lysate with rabbit IgG antiserum to human MPO. The resulting precipitate from HL-60 blasts gave a major band of radioactivity of approximately 77,000 daltons and another band at approximately 46,000 daltons on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The MPO identity of the labeled bands was confirmed by cold competition. The relative mRNA activity expressed as a percentage of radioactivity incorporated into MPO (77,000-dalton band) as compared with total trichloracetic acid (TCA) precipitable radioactivity was 0.2%. Negligible mRNA activity for MPO was present in HL-60 granulocytes, HL-60 macrophages, and normal human granulocytes. Pulse- chase experiments showed that MPO was an approximate 75,000-dalton major band and 77,000-dalton minor band of radioactivity after HL-60 blasts were labeled for 1/2 hour with 35S-methionine and the cell lysate immunoprecipitated and subjected to SDS-PAGE. The chase experiments (one to 24 hours) showed that the 77,000- and 75,000-dalton bands of radioactivity were replaced with two major bands (55,000 and 15,000 daltons) and one minor band (approximately 39,000 daltons) of radioactivity. Six-hour 35S-methionine labeling experiments showed that the relative rate of MPO synthesis compared with total TCA precipitable radioactivity was 0.5% in HL-60 blasts and almost negligible in HL-60 macrophages and granulocytes, normal human granulocytes, and B- lymphocytes. The KG-1 myeloblasts and KG-1a early myeloblasts synthesized a small amount of the 75,000-dalton MPO protein. Although HL-60 cells no longer synthesized MPO after differentiation, HL-60 granulocytes and HL-60 macrophages continued to contain MPO as measured by enzyme activity.(ABSTRACT TRUNCATED AT 400 WORDS)

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