Isolation and identification of a new intracellular antimicrobial peptide produced by Paenibacillus alvei AN5

Alkotaini, Bassam; Anuar, Nurina; Kadhum, Abdul Amir Hassan; Sani, Asmahani Azira Abdu

doi:10.1007/s11274-013-1558-z

Cited by 9 publications

(8 citation statements)

References 31 publications

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“…Molecular identification of EAGSL isolate : One of the isolated colonies, named EAGSL (Electro‐Active Great Salt Lake), was grown on fresh SRB medium for 48 h. The cells were harvested, and the 16S rRNA gene was amplified by PCR as described previously by using forward primer RibS73sp AGAGT TTGAT CCTGG CTCA and reverse primer RibS74sp AAGGA GGTGA TCCAG CCGCA . The PCR product was purified by using a PCR purification kit (Qiagen) prior to DNA sequencing.…”

Section: Methodsmentioning

confidence: 99%

Alginate‐Encapsulated Bacteria for the Treatment of Hypersaline Solutions in Microbial Fuel Cells

et al. 2018

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A microbial fuel cell (MFC) based on a new wild-type strain of Salinivibrio sp. allowed the self-sustained treatment of hypersaline solutions (100 g L , 1.71 m NaCl), reaching a removal of (87±11) % of the initial chemical oxygen demand after five days of operation, being the highest value achieved for hypersaline MFC. The degradation process and the evolution of the open circuit potential of the MFCs were correlated, opening the possibility for online monitoring of the treatment. The use of alginate capsules to trap bacterial cells, increasing cell density and stability, resulted in an eightfold higher power output, together with a more stable system, allowing operation up to five months with no maintenance required. The reported results are of critical importance to efforts to develop a sustainable and cost-effective system that treats hypersaline waste streams and reduces the quantity of polluting compounds released.

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Section: Methodsmentioning

confidence: 99%

Alginate‐Encapsulated Bacteria for the Treatment of Hypersaline Solutions in Microbial Fuel Cells

et al. 2018

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“…The fact that both antimicrobial and anthelmintic activities are expressed by the intracellular extract is not surprising. Most of antimicrobials are expressed outside the cell for cell defense but the intracellular nature of some antimicrobial compounds is not unusual, although their ecological role is still not clear [ 42 , 43 ]. They could be involved in intracellular signalling or have a defensive role [ 44 ].…”

Section: Discussionmentioning

confidence: 99%

Isolation by Miniaturized Culture Chip of an Antarctic bacterium Aequorivita sp. with antimicrobial and anthelmintic activity

Esposito

Ingham²,

Hurtado-Ortiz

et al. 2018

Biotechnology Reports

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HighlightsNovel microbial isolation approach allowed the identification of a Gram-negative Antarctic bacterium belonging to the genus Aequorivita.Aequorivita sp. showed antimicrobial and anthelmintic activity without toxic effect towards eukaryotic cells.The whole genome of Aequorivita sp. was sequenced and compared with other strains to identify biosynthetic gene clusters.This novel approach represents a promising strategy to isolate rare or novel strains useful for biotechnological applications.

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“…SDS-PAGE was performed by electrophoresis, at a constant voltage of 100 V, when the bands entered the separation gel, we adjusted the voltage to 120 V for electrophoresis. Following the run, the protein bands in the gel were stained with Coomassie brilliant blue dye R-250 for 30 min, the SDS-PAGE electrophoresis was over, we used the gel imaging system to photograph the gel and cut the corresponding bands for further liquid chromatography-mass spectrometry analysis [ 73 ].…”

Section: Methodsmentioning

confidence: 99%

Purification, Characterization and Bactericidal Action of Lysozyme, Isolated from Bacillus subtillis BSN314: A Disintegrating Effect of Lysozyme on Gram-Positive and Gram-Negative Bacteria

et al. 2023

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In the present study, lysozyme was purified by the following multi-step methodology: salt (ammonium sulfate) precipitation, dialysis, and ultrafiltration. The lysozyme potential was measured by enzymatic activity after each purification step. However, after ultrafiltration, the resulting material was considered extra purified. It was concentrated in an ultrafiltration centrifuge tube, and the resulting protein/lysozyme was used to determine its bactericidal potential against five bacterial strains, including three gram-positive (Bacillus subtilis 168, Micrococcus luteus, and Bacillus cereus) and two gram-negative (Salmonella typhimurium and Pseudomonas aeruginosa) strains. The results of ZOI and MIC/MBC showed that lysozyme had a higher antimicrobial activity against gram-positive than gram-negative bacterial strains. The results of the antibacterial activity of lysozyme were compared with those of ciprofloxacin (antibiotic). For this purpose, two indices were applied in the present study: antimicrobial index (AMI) and percent activity index (PAI). It was found that the purified lysozyme had a higher antibacterial activity against Bacillus cereus (AMI/PAI; 1.01/101) and Bacillus subtilis 168 (AMI/PAI; 1.03/103), compared to the antibiotic (ciprofloxacin) used in this study. Atomic force microscopy (AFM) was used to determine the bactericidal action of the lysozyme on the bacterial cell. The purified protein was further processed by gel column chromatography and the eluate was collected, its enzymatic activity was 21.93 U/mL, while the eluate was processed by native-PAGE. By this analysis, the un-denatured protein with enzymatic activity of 40.9 U/mL was obtained. This step shows that the protein (lysozyme) has an even higher enzymatic potential. To determine the specific peptides (in lysozyme) that may cause the bactericidal potential and cell lytic/enzymatic activity, the isolated protein (lysozyme) was further processed by the SDS-PAGE technique. SDS-PAGE analysis revealed different bands with sizes of 34 kDa, 24 kDa, and 10 kDa, respectively. To determine the chemical composition of the peptides, the bands (from SDS-PAGE) were cut, enzymatically digested, desalted, and analyzed by LC-MS (liquid chromatography-mass spectrometry). LC-MS analysis showed that the purified lysozyme had the following composition: the number of proteins in the sample was 56, the number of peptides was 124, and the number of PSMs (peptide spectrum matches) was 309. Among them, two peptides related to lysozyme and bactericidal activities were identified as: A0A1Q9G213 (N-acetylmuramoyl-L-alanine amidase) and A0A1Q9FRD3 (D-alanyl-D-alanine carboxypeptidase). The corresponding protein sequence and nucleic acid sequence were determined by comparison with the database.

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Isolation and identification of a new intracellular antimicrobial peptide produced by Paenibacillus alvei AN5

Cited by 9 publications

References 31 publications

Alginate‐Encapsulated Bacteria for the Treatment of Hypersaline Solutions in Microbial Fuel Cells

Alginate‐Encapsulated Bacteria for the Treatment of Hypersaline Solutions in Microbial Fuel Cells

Isolation by Miniaturized Culture Chip of an Antarctic bacterium Aequorivita sp. with antimicrobial and anthelmintic activity

Purification, Characterization and Bactericidal Action of Lysozyme, Isolated from Bacillus subtillis BSN314: A Disintegrating Effect of Lysozyme on Gram-Positive and Gram-Negative Bacteria

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