scholarly journals Characterization and Antibacterial Activity of 7S and 11S Globulins Isolated from Cowpea Seed Protein

Molecules ◽  
2019 ◽  
Vol 24 (6) ◽  
pp. 1082 ◽  
Author(s):  
Seham Abdel-Shafi ◽  
Abdul-Raouf Al-Mohammadi ◽  
Ali Osman ◽  
Gamal Enan ◽  
Samar Abdel-Hameid ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Molecular Mass ◽  
Klebsiella Pneumonia ◽  
Listeria Ivanovii ◽  
Sds Page ◽  
Cell Components ◽  
Cowpea Seed ◽  
11S Globulin ◽  
7S And 11S Globulins ◽  
11S Globulins

The present work was carried out to determine the characteristics and antibacterial activity of 7S and 11S globulins isolated from cowpea seed (Vigna unguiculata (L.) Walp.). The molecular mass of 7S globulin was demonstrated by SDS-PAGE bands to be of about 132, 129 and 95 kDa corresponding the α/, α and β subunits. The molecular mass of 11S globulin was demonstrated by SDS-PAGE bands to be existed between 28 and 52 kDa corresponding the basic and acidic subunits. The minimum inhibitory concentrations MICs of 7S and 11S globulins isolated from cowpea seed were determined against Gram positive bacteria viz: Listeria monocytogenes LMG 10470, Listeria ivanovii FLB 12, Staphylococcus aureus ATCC 25923 and Streptococcus pyogenes ATCC 19615, and Gram negative bacteria such as Klebsiella pneumonia ATCC 43816, Pseudomonas aeruginosa ATCC 26853, Escherichia coli ATCC 25922 and Salmonella ATCC 14028 using disc diffusion assay; they were showed to be in the range 10 to 200 µg/mL. Transmission electron microscope (TEM) examination of the protein-treated bacteria showed the antibacterial action of 11S globulin against S. typhimurium and P. aeruginosa was manifested by signs of cellular deformation, partial and complete lysis of cell components. Adding 11S globulin at both concentrations 50 and 100 µg/g to minced meat showed considerable decreases in bacterial counts of viable bacteria, psychrotrophs and coliforms compared to controls during 15 days storage at 4 °C, reflecting a promising perspective to use such globulin as a meat bio-preservative.

scholarly journals Ultrasound-Assisted Mild Heating Treatment Improves the Emulsifying Properties of 11S Globulins

Molecules ◽  
2020 ◽  
Vol 25 (4) ◽  
pp. 875
Author(s):  
Linlin Liu ◽  
Jianhua Zeng ◽  
Bingyu Sun ◽  
Na Zhang ◽  
Yinyuan He ◽  
...  
Keyword(s):  
Activity Index ◽  
Uv Spectroscopy ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Random Coil ◽  
Control Group ◽  
Emulsifying Properties ◽  
Sds Page ◽  
11S Globulin ◽  
11S Globulins

Ultrasonic technology is often used to modify proteins. Here, we investigated the effects of ultrasound alone or in combination with other heating methods on emulsifying properties and structure of glycinin (11S globulin). Structural alterations were assessed with Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS–PAGE), intrinsic fluorescence spectroscopy, ultraviolet (UV) absorption spectroscopy, and Fourier transform infrared (FTIR) spectroscopy. The size distribution and zeta-potential of 11S globulin were evaluated with a particle size analyzer. An SDS-PAGE analysis showed no remarkable changes in the primary structure of 11S globulin. Ultrasound treatment disrupted the 11S globulin aggregates into small particles with uniform size, narrowed their distribution and increased their surface charge density. Fluorescent spectroscopy and second-derivative UV spectroscopy revealed that ultrasound coupled with heating induced partial unfolding of 11S globulin, increasing its flexibility and hydrophobicity. FTIR further showed that the random coil and α-helix contents were higher while β-turn and β-sheet contents were lower in ultrasound combined with heating group compared to the control group. Consequently, the oil-water interface entirely distributed protein and reduced the surface tension. Moreover, ultrasound combined with heating at 60 °C increased the emulsifying activity index and emulsifying stability index of 11S globulins by 6.49-folds and 2.90-folds, respectively. These findings suggest that ultrasound combined with mild heating modifies the emulsification properties of 11S globulin.

Detection of both Type 1 and Type 2 Plasminogen Activator Inhibitors in Human Monocytes

1990 ◽  
Vol 63 (01) ◽  
pp. 067-071 ◽  
Author(s):  
Joan C Castellote ◽  
Enric Grau ◽  
Maria A Linde ◽  
Nuria Pujol-Moix ◽  
Miquel LI Rutllant
Keyword(s):  
Molecular Mass ◽  
Plasminogen Activator ◽  
Human Peripheral Blood ◽  
Direct Interaction ◽  
Apparent Molecular Weight ◽  
Fibrinolytic System ◽  
Human Monocytes ◽  
Blood Monocytes ◽  
Single Chain ◽  
Sds Page

SummaryIncreasing evidence suggests the involvement of leukocytes in the fibrinolytic system. Monocytes secrete pro-urokinase (Grau, Thromb Res 1989; 53: 145) and it has been shown that these cells have specific receptors for urokinase and plasminogen (Miles, Thromb Haemostas 1987; 58: 936). The aim of this study was to analyse the presence of plasminogen activator inhibitor(s) in platelet-free suspensions of human peripheral blood monocytes and polymorphonuclear leukocytes (PMN). SDS-PAGE and reverse fibrin autography showed an inhibitory band of 50 kDa in the monocyte extracts (Triton X-100) but not in the PMN extracts. Urokinase (u-PA) was mixed with increasing amounts of monocyte extract for 10 min and the mixtures were added to 125Ifibrin coated wells containing plasminogen. A dose-dependent decrease in the u-PA fibrinolytic activity was observed. The amount of inhibition increased when the monocyte releasates were preincubated with u-PA (40% inhibition after 5 min preincubation and 80% after 15 min), indicating a direct interaction between this activator and an inhibitor(s). After SDS-PAGE of monocyte extracts, immunoblotting and peroxidase staining identified both PAI1 and PAI2, with an apparent molecular weight of 47-50 kDa. Monocyte-associated PAI1 formed complexes with single chain t-PA with a molecular mass 50 kDa higher than the molecular mass of the free PAI1. However, a significant amount of PAI remained unbound to t-PA. This inactive PAI1 could have come from a rapid inactivation of the primary active PAI1. These PAI1 and PAI2 detected in human monocytes may be transcendent in the regulation of the fibrinolytic system.

Extraction and characterization of sea anemones compound and its Anti bacterial and hemolytic studies

2020 ◽  
Vol 10 (4) ◽  
pp. 93-97
Author(s):  
Anil Kumar A ◽  
Raja Sheker K ◽  
Naveen B ◽  
Abhilash G ◽  
Akila CR
Keyword(s):  
Antibacterial Activity ◽  
Focal Point ◽  
Haemolytic Activity ◽  
Klebsiella Pneumonia ◽  
Bacterial Strains ◽  
Sea Anemones ◽  
Marine Plants ◽  
Heteractis Magnifica ◽  
Trididemnum Solidum

Seas assets that give us a variety of characteristic items to control bacterial, contagious and viral ailment and mostly utilized for malignancy chemotherapy practically from spineless creatures, for example, bryozoans, wipes, delicate corals, coelenterates, ocean fans, ocean bunnies, molluscs and echinoderms. In the previous 30 - 40 years, marine plants and creatures have been the focal point of overall endeavours to characterize the regular results of the marine condition. Numerous marine characteristic items have been effectively exceptional to the last phases of clinical preliminaries, including dolastatin-10, a group of peptides disengaged from Indian ocean rabbit, Dollabella auricularia. Ecteinascidin-743 from mangrove tunicate Ecteinascidia turbinata, Didemnins was isolated from Caribbean tunicate Trididemnum solidum and Conopeptides from cone snails (Conus sp.), and a developing number of up-and-comers have been chosen as promising leads for expanded pre-clinical appraisals. Sea anemones possess numerous tentacles containing stinging cells or cnidocytes. The stinging cells are equipped with small organelles known as nematocysts. The two species of sea anemones namely, Heteractis magnificaandStichodactyla haddoni, were collected from Mandapam coastal waters of Ramanathapuram district, Tamilnadu, India. The Nematocyst was collected and centrifuged, and the supernatant was lyophilized and stored for further analysis. The amount of protein from Heteractis Magnifica and Stichodactyla haddoni was estimated. The crude extract has shown haemolytic activity on chicken blood and goat blood. In the antibacterial activity of the sea anemone against six bacterial strains Staphylococcus aureus, Salmonella typhii, Salmonella paratyphii, Klebsiella pneumonia, Vibrio cholerae, Pseudomonas aeruginosa. Antibacterial activity of H. Magnifica and S.haddoni was measured as the radius of the zone of inhibition.

scholarly journals TO STUDY THE ANTIBACTERIAL ACTIVITY OF VARIOUS EXTRACTS OF CLAUSENA DENTATA (WILLD.) ROEM.

2019 ◽  
pp. 69-73
Author(s):  
ANNAMALAI MADURAM ◽  
RAJU KAMARAJ
Keyword(s):  
Antibacterial Activity ◽  
Tamil Nadu ◽  
Pathogenic Bacteria ◽  
Volatile Oil ◽  
Stem Bark ◽  
Salmonella Typhi ◽  
Chloroform Extract ◽  
Klebsiella Pneumonia ◽  
Human Pathogens ◽  
Human Pathogenic Bacteria

Objectives: The objectives of the study were to study the antibacterial activity for the various extracts of Clausena dentata against human pathogens. Clausena (Rutaceae) is a genus of about 23 species of unarmed trees and shrubs. The stem bark of C. dentata is used in veterinary medicine for the treatment of wounds and sprains. Even though C. dentata has a lot of potential medical uses, the study of microbiological properties is very scarce. Methods: The plant C. dentata was collected from Kadagaman, near Tiruvannamalai, Tamil Nadu, India, and authenticated by Centre for Advanced Study in Botany, University of Madras, Chennai. The dry powder of stem bark was extracted with hexane, chloroform, and methanol. The extracts were subjected to qualitative phytochemical screening and antibacterial activity against human pathogenic bacteria such as Escherichia coli, Salmonella Typhi, Klebsiella pneumonia, Vibrio cholerae, and Staphylococcus aureus and compared with ciprofloxacin. Results: Qualitative chemical tests revealed the presence of various phytochemicals such as alkaloids, glycosides, carbohydrate, proteins and amino acids, phytosterols, and volatile oil. The antibacterial activity result reveals that all the extracts were are more active against V. cholerae. The activity against Pseudomonas aeruginosa was mild. Conclusion: The activity against V. cholerae was comparable with that of 5 μg/mL ciprofloxacin at the concentration of C. dentata 40 μg/mL. The orders of antibacterial activity against human pathogenic bacteria are hexane, methanol, and chloroform extract of C. dentata.

scholarly journals Identification and characterization of Sulfolobus solfataricusD-gluconate dehydratase: a key enzyme in the non-phosphorylated Entner–Doudoroff pathway

2005 ◽  
Vol 387 (1) ◽  
pp. 271-280 ◽  
Author(s):  
Seonghun KIM ◽  
Sun Bok LEE
Keyword(s):  
Molecular Mass ◽  
Gel Filtration ◽  
Maximal Activity ◽  
Genome Database ◽  
Sds Page ◽  
Key Enzyme ◽  
Bivalent Metal Ions ◽  
Ammonium Sulphate Fractionation ◽  
Identification And Characterization

The extremely thermoacidophilic archaeon Sulfolobus solfataricus utilizes D-glucose as a sole carbon and energy source through the non-phosphorylated Entner–Doudoroff pathway. It has been suggested that this micro-organism metabolizes D-gluconate, the oxidized form of D-glucose, to pyruvate and D-glyceraldehyde by using two unique enzymes, D-gluconate dehydratase and 2-keto-3-deoxy-D-gluconate aldolase. In the present study, we report the purification and characterization of D-gluconate dehydratase from S. solfataricus, which catalyses the conversion of D-gluconate into 2-keto-3-deoxy-D-gluconate. D-Gluconate dehydratase was purified 400-fold from extracts of S. solfataricus by ammonium sulphate fractionation and chromatography on DEAE-Sepharose, Q-Sepharose, phenyl-Sepharose and Mono Q. The native protein showed a molecular mass of 350 kDa by gel filtration, whereas SDS/PAGE analysis provided a molecular mass of 44 kDa, indicating that D-gluconate dehydratase is an octameric protein. The enzyme showed maximal activity at temperatures between 80 and 90 °C and pH values between 6.5 and 7.5, and a half-life of 40 min at 100 °C. Bivalent metal ions such as Co2+, Mg2+, Mn2+ and Ni2+ activated, whereas EDTA inhibited the enzyme. A metal analysis of the purified protein revealed the presence of one Co2+ ion per enzyme monomer. Of the 22 aldonic acids tested, only D-gluconate served as a substrate, with Km=0.45 mM and Vmax=0.15 unit/mg of enzyme. From N-terminal sequences of the purified enzyme, it was found that the gene product of SSO3198 in the S. solfataricus genome database corresponded to D-gluconate dehydratase (gnaD). We also found that the D-gluconate dehydratase of S. solfataricus is a phosphoprotein and that its catalytic activity is regulated by a phosphorylation–dephosphorylation mechanism. This is the first report on biochemical and genetic characterization of D-gluconate dehydratase involved in the non-phosphorylated Entner–Doudoroff pathway.

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.

Isolation and purification of 7S and 11S globulins from broad beans and peas

1990 ◽  
Vol 38 (1) ◽  
pp. 92-95 ◽  
Author(s):  
V. V. Suchkov ◽  
I. A. Popello ◽  
V. Ya. Grinberg ◽  
V. B. Tolstoguzov
Keyword(s):  
Isolation And Purification ◽  
Broad Beans ◽  
7S And 11S Globulins ◽  
11S Globulins

scholarly journals Assessment of Microbial Load of Un-pasteurized Fruit Juices and in vitro Antibacterial Potential of Honey Against Bacterial Isolates

2015 ◽  
Vol 9 (1) ◽  
pp. 26-32 ◽  
Author(s):  
Muhammad Naeem Iqbal ◽  
Aftab Ahmad Anjum ◽  
Muhammad Asad Ali ◽  
Firasat Hussain ◽  
Shahzad Ali ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Viable Count ◽  
Fruit Juices ◽  
Microbial Load ◽  
Resistant Bacteria ◽  
Klebsiella Pneumonia ◽  
Street Vendors ◽  
Permissible Limits ◽  
Development Of Resistance

The development of resistance in bacteria against commonly used antibiotics/drugs is of considerable medical significance. Aim of this study was to determine the microbial load of un-pasteurized packed fruit juices sold in Lahore city and to determine antibacterial activity of five different honey samples against isolated bacteria. Unpasteurized fruit juice samples (n=60) were collected from street vendors. All the samples were subjected to Total viable count (TVC), Staphylococcal count (SC) and Coliform count (CC). One hundred and ten strains of bacteria were isolated from various fruit juices and identified on the basis of cultural characters, morphology and biochemical characters. Mean TVCs, SCs and CCs of juices (6.80±1.91, 5.45±1.06 and 3.25±1.25 log10 CFU/ml respectively) were non-significant with standard permissible limits (p<0.05). Among all the fruit juices, 66.66% of samples had TVC more than 4 log10 CFU/ml, 51.66% of samples had SC more than 3 log10 CFU/ml and 46.66% of samples had CC more than 2 log10 CFU/ml. Among the bacillus isolates purified, were Bacillus alvei, Bacillus subtilis, Bacillus polymyxa, Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumonia, Escherichia coli and Enterobecter. All five different types of honey samples used in this study showed antibacterial activity against B. alvei, B. polymyxa, B. subtilis and S. aureus and no activity against P. aeruginosa, K. pneumonia, Enterobecter and E. coli. It is concluded that microbial load in unpasteurized fruit juices is significantly higher than standard permissible limits which insinuates its possible role in spoilage and food borne illnesses. Periodic monitoring of packed fruit juices should be carried out to make them safe for consumption. Honey can be used as an alternative for treatment of various infections, especially those caused by antibiotic resistant bacteria.

scholarly journals Heparin- and Zn2+-binding proteins from boar seminal plasma.

1999 ◽  
Vol 46 (4) ◽  
pp. 935-939 ◽  
Author(s):  
D Hołody ◽  
J Strzezek
Keyword(s):  
Amino Acid ◽  
Affinity Chromatography ◽  
Molecular Mass ◽  
Seminal Plasma ◽  
Binding Proteins ◽  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Heparin Binding ◽  
Sds Page ◽  
Protein Components

Low molecular mass, heparin-binding proteins from seminal plasma play an important role in gametes interaction whereas plasmatic Zn2+-binding proteins stabilize chromatin and plasmalemma structures and protect spermatozoa in the female reproductive tract. By means of affinity chromatography the heparin- and Zn2+-binding proteins were isolated from boar seminal plasma and both preparations were analyzed by reverse HPLC. Most of the proteins bound to heparine and Zn2+-ions were classified as spermadhesins. Three fractions binding exclusively Zn2+ were isolated. They differ in amino-acid composition, content of glucosamine and content of protein components revealed by SDS/PAGE.

Sign in / Sign up

Export Citation Format

Share Document