scholarly journals Production and Extraction Of Antibacterial Bacteriocin from Pediococcus sp. NWD 015

2015 ◽  
Vol 11 (2) ◽  
Author(s):  
N. Nofisulastri ◽  
Zaenal Bachruddin ◽  
Eni Harmayani
Keyword(s):  
Heat Treatment ◽  
Molecular Weight ◽  
Staphylococcus Aureus ◽  
Listeria Monocytogenes ◽  
Enterococcus Faecalis ◽  
Storage Time ◽  
Bacteriocin Production ◽  
Proteinase K ◽  
Bacteriocin Activity

objectives were to study the growth pattern of Pediococcus sp. NWD 015 and bacteriocin activity, extractionand characterization of bacteriocin, and to determine the effect of storage time and temperature on bacteriocinactivity. Results showed that the bacteriocin activity increased during growth and reached the highest activity duringstationary phase. The maximum bacteriocin production reached after incubation of the cell for 12 h at 37oC in TGEbroth and decreased after 96 h incubation. Extraction with adsorbtion-desorbtion method could increased a specificactivity of bacteriocin. Bacteriocin from Pediococcus sp. NWD 015 is inactivated by Proteinase-K; however it is stillactive by heat treatment at 121oC for 15 min and over pH 2 – 11. Bacteriocin of Pediococcus sp. NWD 015 was effectiveagaints Enterococcus faecalis, Staphylococcus aureus, Eschericia coli, Listeria monocytogenes but not against Salmonellathypimurium. The molecular weight of bacteriocin is 4.95 kDa.Keywords : Bacteriocins, Pediococcus sp NWD 015.

Detection and partial characterization of extracellular inducers of persistence in Staphylococcus epidermidis and Staphylococcus aureus

2021 ◽  
Vol 70 (6) ◽  
Author(s):  
Elyse C. Curry ◽  
Ryan G. Hart ◽  
Danni Y. Habtu ◽  
Neal R. Chamberlain
Keyword(s):  
Molecular Weight ◽  
Staphylococcus Aureus ◽  
Staphylococcus Epidermidis ◽  
Type Species ◽  
Proteinase K ◽  
Partial Characterization ◽  
Content Type ◽  
Link Type ◽  
Inducing Factors

Introduction. This study describes the identification and partial characterization of persistence-inducing factors (PIFs) from staphylococci. Hypothesis/Gap Statement. Increases in persisters during mid-log phase growth indicate that quorum-sensing factors might be produced by staphylococci. Aim. To identify and partially characterize PIFs from Staphylococcus epidermidis RP62A and Staphylococcus aureus SH1000. Methodology. Others have demonstrated a significant increase in persister numbers during mid-log phase. Inducers of this mid-log increase have yet to be identified in staphylococci. Optical density at 600 nm (OD600) was used instead of time to determine when persister numbers increased during logarithmic growth. Concentrated culture filtrates (CCFs) from S. epidermidis and S. aureus were obtained at various OD600s and following incubation at 16 h. The CCFs were used to develop a PIF assay. The PIF assay was used to partially characterize PIF from S. epidermidis and S. aureus for sizing of PIF activity, temperature and protease sensitivity and inter-species communications. Results. The optimal OD600s for S. epidermidis and S. aureus PIF assays were 2.0 and 0.5, respectively. The highest PIF activity for both species was from CCF following incubation overnight (16 h). S. epidermidis ’ PIF activity was decreased by storage at 4 oC but not at 20 oC (16 h), 37 oC (1 h) or 100 oC (15 min). S. aureus ’ PIF activity was decreased following storage at 4 oC (2 weeks) and after boiling at 100 oC for 5 min but not after incubation at 37 oC (1 h). PIF activity from both species went through a 3000 molecular weight cutoff ultrafilter. Proteinase K treatment of S. aureus PIF decreased activity but did not decrease the PIF activity of S. epidermidis . PIF from S. epidermidis did not increase persisters when used to treat S. aureus cells and nor did PIF from S. aureus increase persisters when used to treat S. epidermidis cells. Conclusions. Attempts to discover PIFs for staphylococci were unsuccessful due to the time-based means used to identify mid-log. Both staphylococcal species produce extracellular, low-molecular-weight inducers of persistence when assayed using an OD600 -based PIF assay.

scholarly journals Bacteriocin activity of enterococci and presence of genes related to pathogenesis

2012 ◽  
Vol 30 (No. 4) ◽  
pp. 330-335 ◽  
Author(s):  
K. Trivedi ◽  
R. Sedmíková ◽  
R. Karpíšková
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Activity ◽  
Listeria Monocytogenes ◽  
Virulence Genes ◽  
Bacteriocin Production ◽  
Bacteriocin Activity ◽  
Antimicrobial Substances ◽  
Layer Technique ◽  
Genes Encoding ◽  
The Individual

In total 228 enterococci strains isolated from food were studied. Enterococcus faecalis, E. faecium, and E. casseliflavus were found to be the dominant strains while E. durans and E. mundtii were present in a smaller extent. Antimicrobial activity determined by double layer technique revealed that 150 (65.7%) strains showed antimicrobial activity against the individual tested pathogenic strains of Listeria monocytogenes, Staphylococcus aureus, and methicilin resistant S. aureus (MRSA). Cell-free neutralised supernatants (CFNS) were prepared from 150 potential bacteriocin producers. Of these 150, CFNS 107 (71.3%) strains were active in the bacteriocin production against one or more pathogenic strains tested. S. aureus and MRSA were found to be more sensitive to the antimicrobial substances than L. monocytogenes. Multiplex PCR for the detection of seven virulence genes in bacteriocin producing strains showed that 47.6% of strains were able to amplify one or more virulence genes. E. faecalis was the most virulent species. The presence of tyrdc gene was seen in all bacteriocin producing strains. None of the strains carried genes encoding the resistance to vancomycin.  

scholarly journals 1194. Identification and Characterization of Extracellular Inducers of Persistence in Staphylococcus epidermidis and Staphylococcus aureus

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S619-S619
Author(s):  
Danni Y Habtu ◽  
Neal Chamberlain ◽  
Elyse Curry ◽  
Ryan Hart
Keyword(s):  
Molecular Weight ◽  
Staphylococcus Aureus ◽  
Staphylococcus Epidermidis ◽  
Proteinase K ◽  
Bacterial Cells ◽  
Staphylococcal Species ◽  
Inducing Factors ◽  
Proteinase K Treatment ◽  
Human Infections

Abstract Background This study describes the identification and partial characterization of persistence inducing factors (PIF) from Staphylococcus aureus (S. aureus) and Staphylococcus epidermidis (S. epidermidis). Persistence is an epigenetic process that results in tolerance of bacterial cells to antibiotic treatment, which can result in chronic human infections. Methods Others have demonstrated a significant increase in persister numbers during mid-log phase. Inducers of this mid-log increase have yet to be identified in staphylococci. Optical density at 600 nm (OD600) was used instead of time to determine when persister numbers increased during logarithmic growth. Concentrated culture filtrates (CCF) from S. epidermidis RP62A and S. aureus SH1000 were obtained at various OD600’s and following incubation at 16 h. The CCF’s were used to develop a persistence inducing factor (PIF) assay. The PIF assay was used to partially characterize PIF from S. epidermidis RP62A and S. aureus SH1000 for relative molecular weight, temperature and protease sensitivity and inter-species communications. Results Optimal OD600’s for the S. epidermidis RP62A and S. aureus SH1000 PIF assays were 2.0 and 0.5, respectively. The highest PIF activity for both species was from CCF following incubation overnight (16 h). S. epidermidis RP62A’s PIF activity was decreased by storage at 4o C (2 weeks or longer) but not following incubation at 20o C (16 h), 37o C (1 h) or 100o C (15 min). S. aureus SH1000’s PIF activity was decreased following storage at 4o C (2 week or longer) and after boiling at 100oC for 5 min but not after incubation at 37o C (1 h). PIF activity from both species was less than 3,000 Mrr. Proteinase-K treatment of S. aureus SH1000 PIF decreased activity but did not decrease PIF activity of S. epidermidis RP62A. PIF from S. epidermidis RP62A did not increase persister numbers when used to treat S. aureus SH1000 cells nor did PIF from S. aureus SH1000 increase persister numbers in S. epidermidis RP62A cells. Conclusion Previous attempts to discover PIF’s for staphylococcal species were unsuccessful due to the time-based means used to identify mid-log. Both staphylococcal species appear to produce unique, extracellular, low-molecular-weight inducers of persistence (PIF) when assayed using an OD600-based PIF assay. Disclosures All Authors: No reported disclosures

scholarly journals Purification and characterization of bacteriocins-like inhibitory substances from food isolated Enterococcus faecalis OS13 with activity against nosocomial enterococci

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Ahmed O. El-Gendy ◽  
Dag A. Brede ◽  
Tamer M. Essam ◽  
Magdy A. Amin ◽  
Shaban H. Ahmed ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Antimicrobial Agents ◽  
Food Samples ◽  
Proteinase K ◽  
Clinical Samples ◽  
Bile Salt Hydrolase ◽  
Antibiotic Resistant ◽  
Molecular Weights ◽  
Global Threat

AbstractNosocomial infections caused by enterococci are an ongoing global threat. Thus, finding therapeutic agents for the treatment of such infections are crucial. Some Enterococcus faecalis strains are able to produce antimicrobial peptides called bacteriocins. We analyzed 65 E. faecalis isolates from 43 food samples and 22 clinical samples in Egypt for 17 common bacteriocin-encoding genes of Enterococcus spp. These genes were absent in 11 isolates that showed antimicrobial activity putatively due to bacteriocins (three from food, including isolate OS13, and eight from clinical isolates). The food-isolated E. faecalis OS13 produced bacteriocin-like inhibitory substances (BLIS) named enterocin OS13, which comprised two peptides (enterocin OS13α OS13β) that inhibited the growth of antibiotic-resistant nosocomial E. faecalis and E. faecium isolates. The molecular weights of enterocin OS13α and OS13β were determined as 8079 Da and 7859 Da, respectively, and both were heat-labile. Enterocin OS13α was sensitive to proteinase K, while enterocin OS13β was resistant. Characterization of E. faecalis OS13 isolate revealed that it belonged to sequence type 116. It was non-hemolytic, bile salt hydrolase-negative, gelatinase-positive, and sensitive to ampicillin, penicillin, vancomycin, erythromycin, kanamycin, and gentamicin. In conclusion, BLIS as enterocin OS13α and OS13β represent antimicrobial agents with activities against antibiotic-resistant enterococcal isolates.

scholarly journals Characterization of non-typable strains ofStaphylococcus aureusfrom cases of hospital infection

1987 ◽  
Vol 99 (1) ◽  
pp. 191-200 ◽  
Author(s):  
Ana Vindel ◽  
Cecilia Martín-Bourgon ◽  
Juan A. Saez-Nieto ◽  
Saez Nieto
Keyword(s):  
Heat Treatment ◽  
Staphylococcus Aureus ◽  
Thermal Shock ◽  
Hospital Infection ◽  
Phage Typing ◽  
Alternative Methods ◽  
Sporadic Cases

SUMMARYA high percentage of non-typable (NT)Staphylococcus aureusstrains was isolated in Spanish hospitals during 1984 and 1985. Several alternative methods of typing were employed to study these isolates. These were: phage-typing at 1000 × RTD, phage-typing after heat-treatment (48 °C), thermal shock (56 °C), reverse-typing and induction of additional phages. Using these methods the number of NT isolates was reducedby 60%. Best results were obtained with heat-treatment. Additional phages and reverse-typing were also useful.A scheme for the study of outbreaks and sporadic cases caused by NT strains is proposed using the methods described.

scholarly journals Inhibitors of Pantothenate Kinase: Novel Antibiotics for Staphylococcal Infections

2003 ◽  
Vol 47 (6) ◽  
pp. 2051-2055 ◽  
Author(s):  
Anthony E. Choudhry ◽  
Tracy L. Mandichak ◽  
John P. Broskey ◽  
Richard W. Egolf ◽  
Cynthia Kinsland ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Staphylococcus Aureus ◽  
Biosynthetic Pathway ◽  
Coenzyme A ◽  
Low Molecular Weight ◽  
Pantothenate Kinase ◽  
Staphylococcal Infections ◽  
Low Molecular Weight Compounds ◽  
Novel Antibiotics

ABSTRACT Pantothenate kinase (CoaA) catalyzes the first step of the coenzyme A biosynthetic pathway. Here we report the identification of the Staphylococcus aureus coaA gene and characterization of the enzyme. We have also identified a series of low-molecular-weight compounds which are effective inhibitors of S. aureus CoaA.

Characterization of a Bacteriocin Produced by Enterococcus faecalis N1-33 and Its Application as a Food Preservative

2009 ◽  
Vol 72 (3) ◽  
pp. 524-530 ◽  
Author(s):  
TOMOMI HATA ◽  
MELAKU ALEMU ◽  
MIHO KOBAYASHI ◽  
CHISE SUZUKI ◽  
SUNEE NITISINPRASERT ◽  
...  
Keyword(s):  
Enterococcus Faecalis ◽  
Marked Reduction ◽  
Pathogenic Bacteria ◽  
Culture Supernatant ◽  
Amino Acid Residues ◽  
Bacteriocin Activity ◽  
Food Preservative ◽  
Fermented Bamboo Shoot ◽  
Potential Use

A bacteriocin-producing strain, N1-33, isolated from fermented bamboo shoot was identified as Enterococcus faecalis. The pH-adjusted culture supernatant of this strain consisted of several peptides with bacteriocin activity, and the supernatant inhibited the growth of pathogenic bacteria such as Listeria monocytogenes. The major peptide with bacteriocin activity was purified, and the first 39 amino acid residues of the bacteriocin were found to be identical to enterocin MR10A produced by E. faecalis MRR10-3. Addition of the pH-adjusted and concentrated culture supernatant of strain N1-33 caused a marked reduction in the growth of Bacillus cereus in custard cream and L. monocytogenes in pickled cucumber. These results suggest the potential use of the bacteriocin produced by strain N1-33 in food biopreservation.

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