scholarly journals Hemolysin, Protease, and EPS Producing PathogenicAeromonas hydrophilaStrain An4 Shows Antibacterial Activity against Marine Bacterial Fish Pathogens

2010 ◽  
Vol 2010 ◽  
pp. 1-9 ◽  
Author(s):  
Anju Pandey ◽  
Milind Naik ◽  
Santosh Kumar Dubey
Keyword(s):  
Antibacterial Activity ◽  
Aeromonas Hydrophila ◽  
Pathogenic Bacteria ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Antibacterial Properties ◽  
Cell Extract ◽  
Extracellular Proteins ◽  
Butylated Hydroxytoluene ◽  
Crude Cell Extract

A pathogenicAeromonas hydrophilastrain An4 was isolated from marine catfish and characterized with reference to its proteolytic and hemolytic activity along with SDS-PAGE profile (sodium dodecyl sulphate-Polyacrylamide gel electrophoresis) of ECPs (extracellular proteins) showing hemolysin (approximately 50 kDa). Agar well diffusion assay using crude cell extract of the bacterial isolate clearly demonstrated antibacterial activity against indicator pathogenic bacteria,Staphylococcus arlettaestrain An1,Acinetobactersp. strain An2,Vibrio parahaemolyticusstrain An3, andAlteromonas aurentiaSE3 showing inhibitory zone >10 mm well comparable to common antibiotics. Further GC-MS analysis of crude cell extract revealed several metabolites, namely, phenolics, pyrrolo-pyrazines, pyrrolo-pyridine, and butylated hydroxytoluene (well-known antimicrobials). Characterization of EPS using FTIR indicated presence of several protein-related amine and amide groups along with peaks corresponding to carboxylic and phenyl rings which may be attributed to its virulent and antibacterial properties, respectively. Besides hemolysin, EPS, and protease,Aeromonas hydrophilastrain An4 also produced several antibacterial metabolites.

scholarly journals The Biological Activity of Some Extracted Proteins from Bacillus spp. Isolated from Soil Against Pathogenic Bacteria

2020 ◽  
Vol 30 (4) ◽  
pp. 29
Author(s):  
Haneen Ali Ibrahim ◽  
Neihaya Heikmat Zaki
Keyword(s):  
Breast Cancer ◽  
Antibacterial Activity ◽  
Bacillus Subtilis ◽  
Bacillus Cereus ◽  
Cytotoxic Effect ◽  
Pathogenic Bacteria ◽  
Polyacrylamide Gel Electrophoresis ◽  
Extracellular Proteins ◽  
Mcf7 Cells ◽  
Bacillus Spp

Forty soil samples were collected from different regions in Baghdad city. Among 52 isolate, only 38 isolates (73.1 %) belong to Bacillus spp. All bacterial isolates have been submitted to the cultural, microscopical, biochemical examination and VITEK 2 System, Bacillus cereus was predominantly found 21 (55.3%) followed by Bacillus subtilis 11 (28.9%), then Bacillus amyloliquefaciens was 6 (15.8%). Bacillus cereus (B1) and Bacillus subtilis (B16) isolates were with the highest level with the antibacterial activity when selected. Extracellular proteins were extracted and purified with ammonium sulfate (80%) saturation, then submitted to Sepharose-6B gel filtration column. SDS-polyacrylamide gel electrophoresis was used to determine the molecular weights of proteins, and one band appeared to each of Bacillus cereus and Bacillus subtilis with molecular weight 68 and 50 KDa respectively. The results revealed that higher effect of antibacterial activity on Gram positive bacteria, and moderate on Gram negative bacteria except P.aeruginosa with no effect. The antibiofilm effect of the extract found higher effective against S.pyogens, moderate effect against each of (S.aureus, E.coli, S. marcescens, K. spp) and no effect against P.aeruginosa. The results of cytotoxicity showed that the purified extract was a higher cytotoxic effect on breast cancer MCF7 cells at 1000µg/ml concentration, while the crude extract had a higher cytotoxic effect on breast cancer MCF7 cells at 100µg/ml concentration.

scholarly journals Influence of Gemini Surfactants on Biochemical Profile and Ultrastructure of Aspergillus brasiliensis

2019 ◽  
Vol 9 (2) ◽  
pp. 245 ◽  
Author(s):  
Anna Koziróg ◽  
Anna Otlewska ◽  
Magdalena Gapińska ◽  
Sylwia Michlewska
Keyword(s):  
Gemini Surfactants ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Control Sample ◽  
Extracellular Proteins ◽  
Practical Applications ◽  
Aspergillus Brasiliensis ◽  
Cationic Gemini Surfactants ◽  
Wide Range ◽  
Cationic Compounds

In this study, we investigated the activities of hexamethylene-1,6-bis-(N,N-dimethyl-N-dodecylammonium bromide) (C6), pentamethylene-1,5-bis-(N,N-dimethyl-N-dodecylammonium bromide) (C5), and their two neutral analogues: hexamethylene-1,6-bis-(N-methyl-N-dodecylamine) (A6) and pentamethylene-1,5-bis-(N-methyl-N-dodecylamine) (A5) at concentrations of ½ MIC, MIC, and 2 MIC (minimal inhibitory concentration) against hyphal forms of Aspergillus brasiliensis ATCC 16404. Enzymatic profiles were determined using the API-ZYM system. Extracellular proteins were extracted from the mycelia and analyzed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The ultrastructure was evaluated using a transmission electron microscope (TEM). Both groups of surfactants caused changes in the enzyme profiles. Larger changes in the number and concentration of enzymes were noted after the action of non-ionic gemini surfactants, which may have been due to the 100× higher concentration of neutral compounds. Larger differences between the protein profiles of the control sample and the biocide samples were observed following the use of cationic compounds. On the basis of TEM analyses, we found that, with increasing concentrations of compound C6, the mycelium cells gradually degraded. After treatment at 2 MIC, only membranous structures, multiform bodies, and dense electron pellets remained. Based on these results, we concluded that cationic gemini surfactants, in comparison with their non-ionic analogues, could have a wide range of practical applications as active compounds.

scholarly journals THE PHYSIOCHEMICAL AND ANTIBACTERIAL PROPERTIES OF GALACTOMANNAN EDIBLE FILM OF ARENGA PINNATA INCORPORATED WITH ZINGIBER OFFICINALE ESSENTIAL OIL

2018 ◽  
Vol 11 (12) ◽  
pp. 138
Author(s):  
Juliati Br Tarigan ◽  
Irwana Nainggolan ◽  
Jamaran Kaban
Keyword(s):  
Tensile Strength ◽  
Antibacterial Activity ◽  
Pathogenic Bacteria ◽  
Antibacterial Properties ◽  
Zingiber Officinale ◽  
Salmonella Typhi ◽  
Edible Films ◽  
Shigella Dysenteriae ◽  
The Third ◽  
The Fourier Transform

Objective: This study aimed to demonstrate the incorporation of Zingiber officinale essential (ZOE) oil onto galactomannan from Arenga pinnata (GAP) matrix and determined the antibacterial activity of the edible films (EF).Methods: EF was obtained from the incorporation of GAP (0.5, 0.9, or 1.3 g) with ZOE (0.5 or 1 g) using glycerol (0.6 g) and monoglycerol oleic (0.2 g) as a plasticizer.Results: The thickness of the films increased with the increase of GAP and ZOE oil. However, the tensile strength and water permeability decreased in the fourth EF when the ZOE oil was increased. The maximum value of tensile strength and modulus elongation was obtained with the ratio of GAP to ZOE oil at 0.9, 0.5, 4.502 MPa, and 0.0633 MPa, respectively. The Fourier transform infrared spectrum showed that the interaction occurred between GAP and ZOE oil. The EFs showed antibacterial activity against Escherichia coli, Staphylococcus aureus, Shigella dysenteriae, Salmonella typhi, Pseudomonas aeruginosa, Candida albicans, and Saccharomyces cerevisiae with the highest activity in the third EF. The total bacteria amount in a colony decreased until the 5th day compared with the control. Furthermore, the third EF could inhibit oxygen migration with a respiratory quotient of 7.71.Conclusions: This study revealed that EFs from GAP and ZOE could be prepared and have antibacterial activity against several pathogenic bacteria. 

scholarly journals Thermophilic, Reversible γ-Resorcylate Decarboxylase from Rhizobium sp. Strain MTP-10005: Purification, Molecular Characterization, and Expression

2004 ◽  
Vol 186 (20) ◽  
pp. 6855-6863 ◽  
Author(s):  
Masahiro Yoshida ◽  
Nobuhiro Fukuhara ◽  
Tadao Oikawa
Keyword(s):  
Rattus Norvegicus ◽  
Homo Sapiens ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Hypothetical Protein ◽  
Cell Extract ◽  
Amino Acid Residues ◽  
Sequence Identities ◽  
Rhizobium Sp

ABSTRACT We found the occurrence of thermophilic reversible γ-resorcylate decarboxylase (γ-RDC) in the cell extract of a bacterium isolated from natural water, Rhizobium sp. strain MTP-10005, and purified the enzyme to homogeneity. The molecular mass of the enzyme was determined to be about 151 kDa by gel filtration, and that of the subunit was 37.5 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis; in other words, the enzyme was a homotetramer. The enzyme was induced specifically by the addition of γ-resorcylate to the medium. The enzyme required no coenzyme and did not act on 2,4-dihydroxybenzoate, 2,5-dihydroxybenzoate, 3,4-dihydroxybenzoate, 3,5-dihydroxybenzoate, 2-hydroxybenzoate, or 3-hydroxybenzoate. It was relatively thermostable to heat treatment, and its half-life at 50°C was estimated to be 122 min; furthermore, it catalyzed the reverse carboxylation of resorcinol. The values of k cat/Km (mΜ−1 · s−1) for γ-resorcylate and resorcinol at 30°C and pH 7 were 13.4 and 0.098, respectively. The enzyme contains 327 amino acid residues, and sequence identities were found with those of hypothetical protein AGR C 4595p from Agrobacterium tumefaciens strain C58 (96% identity), 5-carboxyvanillate decarboxylase from Sphingomonas paucimobilis (32%), and 2-amino-3-carboxymuconate-6-semialdehyde decarboxylases from Bacillus cereus ATCC 10987 (26%), Rattus norvegicus (26%), and Homo sapiens (25%). The genes (graA [1,230 bp], graB [888 bp], and graC [1,056 bp]) that are homologous to those in the resorcinol pathway also exist upstream and downstream of the γ-RDC gene. Judging from these results, the resorcinol pathway also exists in Rhizobium sp. strain MTP-10005, and γ-RDC probably catalyzes a reaction just before the hydroxylase in it does.

Purification and some properties of the hemolytic toxin aerolysin

1981 ◽  
Vol 59 (6) ◽  
pp. 430-435 ◽  
Author(s):  
J. Thomas Buckley ◽  
L. Nicole Halasa ◽  
Karen D. Lund ◽  
Sheila MacIntyre
Keyword(s):  
Aeromonas Hydrophila ◽  
Culture Supernatant ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Active Components ◽  
Original Culture ◽  
Hemolytic Action ◽  
Hemolytic Toxin ◽  
Salt Fractionation

Aerolysin, the hemolytic toxin produced by Aeromonas hydrophila, has been purified by a combination of salt fractionation, gel filtration, and ion-exchange and hydroxyapatite chromatography. The resulting protein has a molecular weight of 51 500 and appears homogeneous by polyacrylamide gel electrophoresis in sodium dodecyl sulphate. It is free of detectable protease and phospholipase activities. The purified protein can be separated into two active components with pIs of 5.39 and 5.46 by isoelectric focusing. Both components are found in the original culture supernatant indicating that the multiplicity is not due to proteolysis during isolation. Purified aeroiysin is unstable even at 25 °C and its hemolytic action is inhibited by certain reducing agents including ferrous iron and cysteine. It appears to be the only toxin hemolytic to human cells that is produced by A. hydrophila under the conditions described.

Application of metaproteomic analysis for studying extracellular polymeric substances (EPS) in activated sludge flocs and their fate in sludge digestion

2008 ◽  
Vol 57 (12) ◽  
pp. 2009-2015 ◽  
Author(s):  
C. Park ◽  
R. F. Helm
Keyword(s):  
Activated Sludge ◽  
Protein Separation ◽  
Extracellular Polymeric Substances ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Cation Exchange Resin ◽  
Methanosarcina Barkeri ◽  
Extracellular Proteins ◽  
Sludge Digestion ◽  
Sds Page

Metaproteomic analysis, comprising protein separation and identification, was applied to study extracellular proteins in activated sludges and to track their fate in sludge digestion under both anaerobic and aerobic conditions. The complex sludge proteins were first separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and further analysed by liquid chromatography tandem mass spectrometry (LC-MS/MS) to search their identification. Base extraction and cation exchange resin (CER) method were used to extract EPS from sludges at 0, 12 and 30 days of batch digestion. Several important observations were made during this study. Firstly, protein bands were well separated by extraction/SDS-PAGE protocol used in this study. Secondly, numerous protein bands remained after digestion, indicating that these proteins are not easily degradable in sludge digestion. Thirdly, protein bands detected following anaerobic and aerobic digestion differed, suggesting that proteins degraded in two different digestion environments are not the same. Finally, protein bands that emerged distinctively following anaerobic digestion was found to be subunits of methyl-coenzyme M reductase, the enzyme involved in methane generation, in Methanosarcina barkeri. These results demonstrated that metaproteomic investigation on activated sludge EPS is useful for studying floc formation in activated sludges and their degradation in various digestion environments.

scholarly journals In Vitro and FTIR Spectroscopy: Local Black Fruit Seed Extract as Antibacterial Aeromonas Hydrophila

2021 ◽  
Vol 15 ◽  
pp. 92-98
Author(s):  
Yori Turu Toja ◽  
Eddy Suprayitno ◽  
Aulanni’am ◽  
Uun Yanuhar
Keyword(s):  
Antibacterial Activity ◽  
Ethyl Acetate ◽  
Aeromonas Hydrophila ◽  
Antibacterial Properties ◽  
Inhibition Zone ◽  
Seed Extract ◽  
Water Fraction ◽  
Natural Treatment ◽  
Ft Ir

Indonesia has many local plants with potential as herbal antibacterial properties, one of which is the local black fruit of Wandama. The objective of this study is to determine the antibacterial activity of the extract of black fruit seed against A. Hydrophila by in vitro and FTIR by Tilapia. The benefit of this research is to make black fruit seed extract as a natural remedy against tilapia attacked by Aeromonas Hydrophila. The antibacterial activity of black fruit seed extract against A. Hydrophila is investigated in vitro and using FTIR on tilapia in this research. The aim of this study is to establish black fruit seed extract as a natural treatment for tilapia infected with Aeromonas Hydrophila. The disc approach was used to conduct the in vitro research. Agar medium was poured into a sterile petri dish, and 2 drops of bacteria from the liquid medium were uniformly distributed and cooled. On the press, disc paper with black fruit seed extract was put and incubated for 18-24 hours at 35 C. An FTS 1000 version spectrophotometer system was used for the FTIR test. For the FT-IR study, a fraction of dried black fruit seeds was used. For clear preparation, 100 mg of dry extract is condensed in KBr pellets. A specimen of loaded black fruit seed samples was analyzed using FT-IR spectroscopy with a scanning range of 400-4000 cm-1 and a resolution of 4 cm-1. The results showed that the ethyl acetate fraction produced an inhibition zone (13.65 mm), the water fraction produced an inhibition zone (11.21 mm), and the n-hexane fraction produced the weakest inhibition zone (11.21 mm) (5.31mm). The concentration test results from 125 ppm, 250 ppm, 500 ppm and 1000 ppm obtained a large inhibition zone at a concentration of 1000 ppm with an inhibition zone area of 11.43 + 0.02. The absorbance value of black fruit seed extract in ethyl acetate solvent at 3430 cm-1 revealed the absorption band with the strongest vibration of the hydroxylate (-OH) functional group, according to FTIR research. Final thoughts with an average value of 13.65 + 0.06, black fruit seed extract using ethyl acetate can inhibit A. Hydrophila bacteria.

scholarly journals Chemical composition and antibacterial activities of the essential oils from Ocotea zahamenensis Van Der Werff (Lauraceae)

2021 ◽  
Vol 16 (1) ◽  
pp. 115-125
Author(s):  
Razafiarimanga Zara Nomentsoa ◽  
Randriamampianina Lovarintsoa Judicael ◽  
Randrianarivo Hanitra Ranjàna ◽  
Ralitera Andrianirina Manampisoa ◽  
Rakoto Danielle Aurore Doll ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Body Weight ◽  
Essential Oils ◽  
Pathogenic Bacteria ◽  
Vibrio Fischeri ◽  
Antibacterial Properties ◽  
Antibacterial Activities ◽  
Plant Parts ◽  
Detection Analysis ◽  
Toxicological Studies

The present work aimed to study the composition and antibacterial properties of the essential oils (EO) of Ocotea zahamenensis leaves (LEO), stem (SEO) and root (REO) barks from two harvest periods (March and June). All EOs were extracted by hydrodistillation from fresh plant parts with yields up to 4.5%. They are colourless, clear, with a strong odour, heavy, levogyre, with a low acid index and an ester index up to 14.89. Gas chromatography/flame ionisation detection analysis of these EOs identified 5 to 12 components representing 96.06 to 99.96% of the overall composition. Safrole was by far the most predominant constituent with contents ranging from 77.45% (SEO, June) to 97.05% (REO, March). The antibacterial activity was tested against eight pathogenic bacteria including 4 Gram (-) and 4 Gram (+) using microdilution assays. With Minimum Inhibitory Concentration (MIC) values of less than 1 mg/mL, all EOs showed antibacterial activity which varied according to the strain. There was not much difference between the activities of March and June Eos, and in both cases SEO were slightly more effective than LEO and REO. All EOs had bacteriostatic action on Bacillus cereus and Vibrio fischeri and bactericidal on almost other strains. When administered orally to mice at 0.5 mg/kg body weight, all EOs caused symptoms of intoxication. Their LD50 varied from 1.019 to 2.73 g/kg body weight. These EOs could be a new source of safrole and could be used for various purposes with further toxicological studies.

scholarly journals Cymbopogon nardus Essential Oil as Protein Inducer in Bacillus subtilis ATCC21332

2018 ◽  
Vol 1 (1) ◽  
Author(s):  
Hairul Shahril Muhamad ◽  
Ismatul Nurul Asyikin Ismail ◽  
Nabilah Ahmad Alhadi ◽  
Salina Mat Radzi ◽  
Maryam Mohamed Rehan ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Bacillus Subtilis ◽  
Essential Oil ◽  
Protein Production ◽  
Antimicrobial Agents ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Extracellular Proteins ◽  
Specific Chemical ◽  
Cymbopogon Nardus

Protein production by bacteria might be increased in stressful conditions such as in the presence of antimicrobial agents. Many studies have proven that antibiotics or antimicrobial agents at low concentration are able to activate or repress gene transcription process in bacteria. However, there have been comparatively few studies on the potential of natural compounds in nature as a specific chemical signal that can trigger a variety of biological functions. An attempt was made to study the effect of essential oil from Cymbopogon nardus in regulating protein production by Bacillus subtilis ATCC21332. The bacterial cells were further exposed to the C. nardus essential oil at concentration of 0.02 % for 48 h at 37°C. The intracellular proteins were then isolated and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Proteins profile showed that a band with approximate size of 180 kDa appeared for the treated bacteria with C. nardus essential oil. An alignment of peptide sequences to the NCBI BLAST database revealed that B. subtilis ATCC21332 in stressful condition tend to produce intracellular protein recognized as respiratory nitrate reductase ? subunit enzyme. Besides, the extracellular proteins secreted by B. subtilis ATCC21332 after being subjected to 0.02% of C. nardus essential oil for 48 and 72 h at 30°C, were further analyzed on antimicrobial activity. The extracellular proteins secreted by B. subtilis ATCC21332 prior to enhancing with 0.02 % C. nardus essential oil at 30°C for 72 h exhibited antimicrobial activity towards two strains of bacteria, which are Bacillus cereus and Escherichia coli.

scholarly journals Antibacterial Activity of Liverworts of Dumortiera hirsute (Sw.) Nees Ethyl Acetate Extract Against Pathogenic Bacteria

2021 ◽  
Vol 9 (2) ◽  
pp. 75
Author(s):  
Luthfiah Luthfiah ◽  
Dwi Setyati ◽  
Sattya Arimurti
Keyword(s):  
Escherichia Coli ◽  
Antibacterial Activity ◽  
Ethyl Acetate ◽  
Pathogenic Bacteria ◽  
Ethyl Acetate Extract ◽  
Antibacterial Properties ◽  
Salmonella Typhi ◽  
Diffusion Method ◽  
E Coli ◽  
Agar Well Diffusion Method

Dumortiera hirsuta is one of the liverworts that can be used as a medicinal to prevent infection by pathogenic bacteria. The content of secondary metabolites of D. hirsuta has potential as antibacterial properties includes flavonoids, alkaloids and steroids. This research is to analyze the antibacterial activity of moss D. hirsuta against pathogenic bacteria that will be beneficial to humans. Liverworts of D. hirsuta were extracted using ethyl acetate solvent and tested against three types of pathogenic bacteria using the agar well-diffusion method. The results of this study indicated that the ethyl acetate extract of D. hirsuta at 100% concentration could inhibit the growth of Escherichia coli, Staphylococcus aureus, and Salmonella typhi bacteria. The range of antibacterial activity categories of the ethyl acetate extract of D. hirsuta to E. coli, S. aureus, and S. typhi between weak to moderate.

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