Bacillus subtilis-based microbial cell factories

Trabelsi, Heykel; Dhali, Debarun; Yaseen, Yazen; Leclère, Valérie; Jacques, Philippe; Coutte, François

doi:10.1016/b978-0-12-821477-0.00002-7

Cited by 2 publications

(2 citation statements)

References 223 publications

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“…Among all bacteria, the genus Bacillus is considered the most suitable for the mass production of lysozyme [ 28 , 29 , 30 , 31 ], while among all Bacillus spp., Bacillus subtilis is the most popular species for higher lysozyme production. This is due to its excellent fermentation properties and tremendous ability to secrete proteins (lysozyme) directly into the growth medium, which offers great advantages in downstream processing, and it is nonpathogenic and free of endotoxins [ 29 , 32 , 33 ].…”

Section: Introductionmentioning

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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Section: Introductionmentioning

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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“…The RBS sequence, extending from the SD sequence to the first 5-6 codons of the coding sequence, plays a crucial role in translation efficiency and fidelity, thereby directly affecting protein abundance and quality (Asahara et al, 2021;Faure et al, 2016;Trabelsi et al, 2021). The SD sequence is particularly influential in this context (Asahara et al, 2021).…”

Section: Rpsu and Rpsb May Influence Sd Sequence Preferencementioning

confidence: 99%

Adaptive strategies of Listeria monocytogenes : Evolved variants with altered stress resistance and fitness

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Ribosomal mutations enable a switch between high fitness and high stress resistance in Listeria monocytogenes 3 A single point mutation in the Listeria monocytogenes ribosomal gene rpsU enables SigB activation independently of the stressosome and the anti-sigma factor antagonist RsbV 4 Stress resistant rpsU variants of Listeria monocytogenes can become underrepresented due to enrichment bias 5 Activation of a silent lactose utilization pathway in an evolved Listeria monocytogenes F2365 outbreak isolate

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Bacillus subtilis-based microbial cell factories

Cited by 2 publications

References 223 publications

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

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

Adaptive strategies of Listeria monocytogenes : Evolved variants with altered stress resistance and fitness

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