Antimicrobial activity of bacteriocin isolated and purified from rumen liquor collected from slaughtered goats

2015 ◽  
Vol 49 (6) ◽  
Author(s):  
S. Barathiraja ◽  
J. Thanislass ◽  
P. X. Antony ◽  
S. Venkatesaperumal

Bacteriocin like substance with antimicrobial activity was purified from freshly collected rumen liquor using 60% ammonium sulphate precipitation followed by ion exchange(SP-Sepharose) and gel filtration (Sephadex G25) chromatographic techniques. Purity of the product was checked on SDS-PAGE, having molecular weight of 6.5 kDa. Anti-microbial activity was demonstrated using <italic>Bacillus subtilis</italic> by gel overlay method and agar cut well diffusion method. Proteomic analysis confirmed the substance as bacteriocin. The purified sample was resistant to the action of protease. The substance was active at pH 4, 7 and 10. It was also active at autoclave temperature. The peptide purified was found to inhibit the growth of <italic>Staphylococcus aureus</italic> (MTCC87), <italic>Listeria monocytogenes</italic> (MTCC 657) and <italic>Pseudomonas aeurginosa</italic> (MTCC 424).

2015 ◽  
Vol 1 (2) ◽  
pp. 126
Author(s):  
Urnemi ◽  
Sumaryati Syukur ◽  
Endang Purwati ◽  
Sanusi Ibrahim ◽  
Jamsari

ABSTRACT Lactic acid bacteria (LAB) were isolated from of cocoa beans fermentation Forestero variety from West Sumatera, that were eleven isolates. The isolates were tested to antimicrobial activity against pathogenic bacteria E.coli NBRC 14237, Staphylococcus aureus NBRC 13276, Bacillus subtilis BTCCB 612, listeria m. dan S. Typhii. Results the research showed that, isolates had inhibition zone to pathogenic bacteria, that were 7 mm till 12 mm at 48 hours observation. R2.4 isolate was most potential to inhibition zones growth pathogenic bacteria, that was 11mm till 12 mm to five pathogens. R2.4 isolates was the highest to against pathogenic bacteria (Bacillus subtilis BTCCB, Listeria monocytogenesis and Staphylococcus aureus NBRC) had inhibition zones, that was 12.00 mm till 48 hours. Listeria monocytogenesis had been known as pest bacterium of food born, so that R2.4 isolate can be used as food biopreservative. Crude of R2.4 isolate molecular weight was 10 kDa by SDS-PAGE.  Key words: Lactic acid bacteria, Antimicrobial activity, SDS-PAGE, Cocoa fermentation and food biopreservative                                                      


1999 ◽  
Vol 67 (8) ◽  
pp. 4014-4018 ◽  
Author(s):  
Hisaaki Sato ◽  
Takao Watanabe ◽  
Yasuko Murata ◽  
Ayumi Ohtake ◽  
Mayumi Nakamura ◽  
...  

ABSTRACT A new serotype of Staphylococcus hyicus exfoliative toxin (SHET), serotype B, was isolated from the culture filtrate of a plasmid-carrying strain of S. hyicus. The new SHET was purified by precipitation with 70% saturated ammonium sulfate, gel filtration on a Sephadex G-75 column, column chromatography on DEAE–Cellulofine A-500, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The new SHET caused exfoliation of the epidermis as determined by the so-called Nikolsky sign when inoculated into 1-day-old chickens. The new SHET was serologically different fromStaphylococcus aureus exfoliative toxins (ETs) (ETA, ETB, and ETC) and from the SHET from the plasmidless strain but showed the same molecular weight as the other serotypes of toxins on SDS-PAGE. It was thermolabile and lost its toxicity after being heated at 60°C for 30 min. We propose that the new SHET be designated SHETB and that the SHET produced by the plasmidless strain be designated SHETA.


2020 ◽  
Vol 9 (1) ◽  
pp. 30
Author(s):  
Mochammad Sriduresta Soenarno ◽  
Irma Isnafia Arief ◽  
Cece Sumantri ◽  
Epi Taufik ◽  
Lilis Nuraida

Bakteriosin adalah peptida dengan aktivitas antibakteri yang diproduksi oleh bakteri asam laktat dan digunakan sebagai pengawet alami. Penelitian sebelumnya menunjukkan bahwa Lactobacillus plantarum IIA-1A5 memproduksi bakteriosin yang diberi nama Plantarisin IIA-1A5 pada medium pertumbuhan yang dibuat dari whey yang diperkaya skim. Untuk aplikasi sebagai pengawet alami dan untuk memperbaiki masa simpan dan aktivitas anti mikrobanya, plantarisin perlu dienkapsulasi dan dikeringbekukan. Tujuan dari penelitian ini adalah untuk mengkarakterisasi dan mengevaluasi aktivitas antimikroba dari sediaan plantarisin IIA-1A5 yang terpurifikasi parsial dan terenkapsulasi kering beku. Ekstraksi dan purifikasi dari bakteriosin dimulai dengan presipitasi dengan ammonium sulfat, yang diikuti dengan dialysis, dan penukar kation kromatografi. Purifikasi parsial dari plantarisin kemudian dimikroenkapsulasi dengan maltodextrin kemudian dilanjutkan dengan proses kering beku. Berdasarkan pada SDS-PAGE, fraksi protein ke-7 (F7) dari plantarisin yang dipurifikasi parsial memiliki pita tunggal dan berat molekul sekitar 9,65 kDa. Konfirmasi lebih lanjut dengan menggunakan MALDI-TOF MS, ternyata pita tunggal tersebut terdiri dari 5 peptida yang diidentifikasi berbobot molekul masing-masing sebagai berikut 5,5, 7,80, 7,96, 9,09, dan 9,27 kDa. Plantarisin kering beku memiliki aktivitas antimikroba terhadap Staphylococcus  aureus tiga kali lipat dibandingkan dengan aktivitas antimikroba dari supernatan bebas sel, dan lebih tinggi dibandingkan dengan nisin, namun kurang bila dibandingkan dengan antibiotik ampisilin dan penisilin. Kesimpulannya, aktivitas antimikroba plantarisin kering beku dapat ditentukan dan lebih tinggi dibandingkan dengan nisin, ampisilin dan penisilin.Characterization of Plantarisin IIA-1A5 as Antimicrobial subtances and Evaluation of Acitivity of Freeze-dried Microencapsulated PreparationAbstractBacteriocins are peptides with antibacterial activity produced by lactic acid bacteria and used as natural preservatives. Previous studies showed that Lactobacillus plantarum IIA-1A5 produces bacteriocin named plantaricin IIA-1A5 in the medium consisting whey enriched with skim milk. For application as food preservatives and to improve its shelf-lie and activity, plantaricin was needed to be microencapsulated and freeze dried. The objective of this research was to characterize and evaluate the activity of partially purified freeze dried microencapsulated plantaricin IIA-1A5. Characterisation of partially purified plantaricin IIA-IA5 includes the identification of active fractions and molecular weight, evaluation of activity at different stage of purification and evaluation of antimicrobial activity of freeze dried microencapsulated plantaricin IIA-IA5. Extraction and prificafication of the bacteriocins started with precipitacion with ammonium sulfate, followed by dialysis, and cation exchange chromatography. The partial purified of plantaricin was then microencapsulated in maltodextrin followed by freeze drying. Based on SDS-PAGE, the protein fraction F7 of partially purified plantaricin had a single band and molecular weight about 9.65 kDa. Further analyses using MALDI-TOF, it revealed that five peptides were identified from one single band plantaricin with molecular weight 5.5, 7.80, 7.96, 9.09, and 9.27 kDa, respectively. The freeze dried plantaricin freeze showed antimicrobial activity against Staphylococcus aureus three times stringer as compared to the activity of cell free supernatant, and was higher than nicin, but less than antibiotic ampicilin and penicilin. As concusion, the activity of freeze dried plantaricin could be determined and had a higher value than nicin, ampicilin and penicilin.


1988 ◽  
Vol 34 (7) ◽  
pp. 855-859 ◽  
Author(s):  
Mohinder Kaur ◽  
K. K. Tripathi ◽  
Meenakshi Gupta ◽  
P. K. Jain ◽  
M. R. Bansal ◽  
...  

Conditions are described for the production of extracellular elastase by Bacillus subtilis. The yield of enzyme was maximum in shake–cultures grown in Syncase medium at 37 °C and was stable in culture supernatants. The enzyme, purified by ammonium sulphate precipitation and Sephadex G-75 gel filtration, showed a molecular weight of 25 000 and activity between pH 6.0 and 9.5, with an optimum of 9.0 in Tris–maleate buffer. Elastinolytic activity was maximum in glycine–NaOH buffer and minimum in phosphate buffer. Enzyme activity was adversely affected by temperature ≥ 40 °C.


2009 ◽  
Vol 15 (1) ◽  
pp. 37-44
Author(s):  
Noor Erma Sugijanto ◽  
H. Putra ◽  
F. Pritayuni ◽  
N. Albathaty ◽  
Noor Cholies Zaini

The antimicrobial potential of two endophytic fungi isolated from Alyxia reinwardtii BL, towards selected bacteria (Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Bacillus subtilis FNCC 0059, Pseudomonas aeruginosa ATCC 27853, Salmonella typhi) and fungi (Candida albicans) was tested using ethyl acetate, n-hexane and n-buthanol extracts of fungi cultivated under malt extract liquid fermentation. The extracts were evaluated for its antimicrobial activity by disc diffusion method followed its MIC by agar dilution. Bioautography assay for activity-directed fractionation were also conducted against Bacillus subtilis and Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922. Streptomycin sulphate and myconazole used as reference standards. Lecythophora sp. strain 30.1 and 30.5 have broad spectrum antimicrobial activity and a potential source of new classes of antibiotics that could be useful for medicines and biological control agents.


2015 ◽  
Vol 4 (2) ◽  
pp. 94
Author(s):  
Urnemi ◽  
Sumaryati Syukur ◽  
Endang Purwati ◽  
Sanusi Ibrahim ◽  
Jamsari

 ABSTRACTLactic acid bacteria (LAB) were isolated from cocoa beans fermentation Forestero variety from West Sumatera, that were eleven isolates. The isolates were tested to antimicrobial activity against pathogenic bacteria E.coli NBRC 14237, Staphylococcus aureus NBRC 13276, Bacillus subtilis BTCCB 612, listeria m. dan S. Typhii.Results the research showed that, isolates had inhibition zone to pathogenic bacteria, that were 7 mm till 12 mm at 48 hours observation. R2.4 isolate was most potential to inhibition zones growth pathogenic bacteria, That was 11mm till 12 mm to five pathogens. R2.4 isolates was the highest to against pathogenic bacteria (Bacillus subtilis BTCCB, Listeria monocytogenesis and Staphylococcus aureus NBRC) had inhibition zones, that was 12. mm till 48 hours. Listeria monocytogenesis had been known as pest bacterium of food born, so that R2.4 isolate can be used as food biopreservative. Crude of R2.4 isolate molecular weight was 10 kDa by SDS-PAGE.  Key words: Lactic acid bacteria, antimicrobial activity, SDS-PAGE, cocoa fermentation  and food biopreservative    


2019 ◽  
Vol 9 (02) ◽  
Author(s):  
Hussein A Kadhum ◽  
Thualfakar H Hasan2

The study involved the selection of two isolates from Bacillus subtilis to investigate their inhibitory activity against some bacterial pathogens. B sub-bacteria were found to have a broad spectrum against test bacteria such as Staphylococcus aureus and Pseudomonas aeruginosa. They were about 23-30 mm and less against Klebsiella sp. The sensitivity of some antibodies was tested on the test samples. The results showed that the inhibitory ability of bacterial growth in the test samples using B. subtilis extract was more effective than the antibiotics used.


2019 ◽  
Vol 18 (5) ◽  
pp. 262-274
Author(s):  
E. Benyagoub ◽  
N. Nabbou ◽  
S. Boukhalkhel ◽  
I. Dehini

The medicinal value of the plants is due to their chemical components that bring a definite physiological action on the human body to prevent the diseases. In this work, we investigated the antimicrobial activity of leaves’ extracts of Quercus robur L., collected from the Algerian upper highlands, on ten bacterial strains and one fungal strain known to be pathogenic. First, we performed a qualitative phytochemical analysis, and second, antimicrobial activity tests performed by agar diffusion method (disc and well) with the determination of MIC by broth macro-dilution method. Given the results, it appears that obtained macerates of Quercus robur L. were rich in bioactive phytoconstituents such as alkaloids, anthraquinones, saponins, tannins, and other components. The yield of aqueous and methanolic macerates of leaves was 8.5 ± 1.41 and 22.4 ± 4.36%, respectively. The bacterial resistance was relatively important to several antibiotics, namely, ampicillin, amoxicillin + clavulanic acid for strains of Escherichia coli and Salmonella sp. However, Staphylococcus aureus strains were resistant to fusidic acid, penicillin, and oxacillin; while Enterococcus faecalis was resistant to fusidic acid, penicillin, oxacillin, and ticarcillin. The antibacterial activity of the macerates toward tested microbial strains showed that the aqueous and methanolic macerates of the leaves were proportional to the tested concentration and active not only against Gram-positive and Gram-negative bacteria but also on the fungal species Candida albicans. The estimated MIC for Escherichia coli, Enterococcus faecalis, and Staphylococcus aureus was in the order of 10 mg/mL, which seems more effective than toward Salmonella sp., Klebsiella pneumoniae, Pseudomonas aeruginosa, and Candida albicans which were in the order of 30 mg/mL. These preliminary results confirm that the part of the studied plant had a very good antimicrobial activity that was proportional to the serial concentrations of the tested extracts.


Author(s):  
Iswariya S. ◽  
Uma T. S.

Objective: The present study was designed to identify the bioactive phytochemicals and its antibacterial and in vitro anti-inflammatory potential of aqueous and methanolic seed extract of Citrullus lanatus.Methods: The phytochemical screening of both the aqueous and methanolic seed extract was carried out qualitatively to identify the major Phyto-constituents present in the extracts. The antimicrobial activity of the extracts was evaluated against six pathogenic bacterial strains by agar well diffusion method and the Minimum inhibitory concentration (MIC) was determined by broth dilution method. In vitro anti-inflammatory activity of C. lanatus seed extracts was evaluated by using human red blood cell (HRBC) membrane stabilization and inhibition of albumin denaturation method.Results: The results of the study indicated that both the extracts of the seed having antimicrobial activity, while the methanolic extract showed more significant activity against the tested organism than aqueous extract. Methanol extract had the lowest MIC of 1.562 mg/ml against Escherichia coli, Staphylococcus aureus, Klebsiella pneumonia, Pseudomonas aeruginosa and Bacillus subtilis, whereas in aqueous extract was highly sensitive to Bacillus subtilis, E. coli and Klebsiella pneumonia with MIC of 3.125 and 6.25 mg/ml, respectively. Methanolic extracts exerted comparative higher anti-inflammatory activity than aqueous extract.Conclusion: Present study provides a firm evidence to support that the synergistic effect of C. lanatus seed extracts having potent anti-inflammatory and antimicrobial property, which might serve as an effective drug for various microbial infections and inflammatory disorders.


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