Isolation of a putative virulence agent, cytotoxic serine-elastase, from a newly isolated Pseudomonas aeruginosa ZuhP13

Kotb, Essam; El-Zawahry, Y.A.; Saleh, Ghadeer E

doi:10.1007/s12038-018-9829-3

Cited by 5 publications

(11 citation statements)

References 21 publications

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“…The best reaction pH for gasm32 elastase was determined to be 8.0 (Fig 2 ), which is the same as that of P. aeruginosa [36] and the B. megaterium-TK1 strain isolated from saltwater [37]. This optimal pH was different from those obtained for other elastases in other studies [38], such as the alkaline elastase from Micrococcus luteus (pH 9.3, [26]), B. megaterium (pH 7.5, [29]), P. aeruginosa ZuhP13 (pH 7.5, [13]), and the alkalophilic Bacillus strain Ya-B (pH 11.75, [25]).…”

Section: Plos Onesupporting

confidence: 49%

“…Centrifugation was performed at 5,000 rpm for 20 min using a refrigerated centrifuge (Hermle Labortechnik GmbH, Z 326 K, Germany) at 4˚C to separate the bacterial cells. Supernatants were used for the determination of elastase productivity and protein concentration as described by Kotb et al [13] with slight modifications.…”

Section: Enzyme Production and Protein Measurementmentioning

confidence: 99%

“…Elastase is expected to have potential application in meat tenderization because of its specific hydrolysis toward elastin. Kotb et al [13] reported that among a total of 120 clinical specimens that were collected from the hospitals of Zagazig University (Zagazig, Egypt) from different sources, including pus, urine, wounds, sputum, eyes, blood, and ear infections, only 54 isolates of Pseudomonas aeruginosa ZuhP13 were positive for cytotoxic serine-elastase productivity. Zupetic et al [32] isolated Pseudomonas aeruginosa from 238 unique ICU patients belonging to two tertiary-care centers inside the University of Pittsburgh Medical Center health system, and 75% of the isolates produced elastase activity.…”

Section: Plos Onementioning

confidence: 99%

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A survey of elastase-producing bacteria and characteristics of the most potent producer, Priestia megaterium gasm32

et al. 2023

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Ninety-one elastase-producing bacterial isolates were recovered from different localities of the Eastern Province of Saudi Arabia. Elastase from the best isolate Priestia megaterium gasm32, from luncheon samples was purified to electrophoretic homogeneity using DEAE-Sepharose CL-6B and Sephadex G-100 chromatographic techniques. The recovery was 17.7%, the purification fold was 11.7x, and the molecular mass was 30 kDa. Enzymatic activity was highly repressed by Ba2+ and almost completely lost by EDTA, but it was greatly stimulated by Cu2+ ions, suggesting a metalloprotease type. The enzyme was stable at 45°C and pH 6.0–10.0 for 2 hours. Ca2+ ions considerably enhanced the stability of the heat-treated enzyme. The Vmax and Km against the synthetic substrate elastin–Congo red were 6.03 mg/mL, and 8.82 U/mg, respectively. Interestingly, the enzyme showed potent antibacterial activity against many bacterial pathogens. Under SEM, most bacterial cells showed loss of integrity, damage, and perforation. SEM micrographs also showed a time-dependent gradual breakdown of elastin fibers exposed to elastase. After 3 hours, intact elastin fibers disappeared, leaving irregular pieces. Given these good features, this elastase may be a promising candidate for treating damaged skin fibers with the inhibition of contaminating bacteria.

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Section: Plos Onesupporting

confidence: 49%

Section: Enzyme Production and Protein Measurementmentioning

confidence: 99%

Section: Plos Onementioning

confidence: 99%

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A survey of elastase-producing bacteria and characteristics of the most potent producer, Priestia megaterium gasm32

et al. 2023

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“…Bacterial proteolytic enzymes, especially those produced by non-pathogenic bacteria, have a special concern in many medical and non-medical applications [25]. This…”

Section: Discussionmentioning

confidence: 99%

“…Bacterial proteolytic enzymes, especially those produced by non-pathogenic bacteria, have a special concern in many medical and non-medical applications [ 25 ]. This study evaluated the potentiality of a serine protease from the non-pathogenic bacterium B. subtilis on the healing of burn wounds in mice.…”

Section: Discussionmentioning

confidence: 99%

Histological Studies on a Newly Isolated Bacillus subtilis D10 Protease in the Debridement of Burn Wound Eschars Using Mouse Model

et al. 2021

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Background: Proteases are among the most important industrial enzymes, playing a critical role in the physiological, biochemical, and regulatory processes of all living organisms. This study evaluated the histological effects of a Bacillus subtilis D10 protease in combination with the antibacterial ointment silver sulfadiazine (SSD) on the burned skin of mice. Materials and Methods: The bacterial proteolytic enzyme was produced and purified through DEAE-Sepharose CL-6B and Sephadex G-100 FF. The in vitro protease specificity was then determined. The dorsal skin of albino mice was burned with 80% HCl solution, then treated under three conditions: cold cream, SSD, and SSD combined with the tested protease. After 15 days of daily treatment, the mice were sacrificed and skin tissue samples were histopathologically examined using hematoxylin eosin, and Masson trichrome staining. Results: The D10 protease hydrolyzed the proteinaceous components of eschars (fibrin, normal collagen, and denatured collagen) in vitro. Mice skins treated with protease and SSD mixture showed promising results, with more rapid healing than the other treatments. This group regenerated epidermis and dermis with newly formed granulated follicles, fibroblasts and blood capillaries in the dermis, and collagen fibers in the hypodermis. Conclusions: These results suggest that the serine protease produced by B. subtilis D10 promotes wound healing of mice skin burnt with HCl and restores the normal architectural pattern in a shorter time than the standard treatments.

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A putative cytotoxic serine protease from Salmonella typhimurium UcB5 recovered from undercooked burger

Kotb

El-Nogoumy

Alqahtani

et al. 2023

Sci Rep

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A putative virulence exoprotease designated as UcB5 was successfully purified from the bacterium Salmonella typhimurium to the electrophoretic homogeneity with 13.2-fold and 17.1% recovery by hydrophobic, ion-exchange, and gel permeation chromatography using Phenyl-Sepharose 6FF, DEAE-Sepharose CL-6B, and Sephadex G-75, respectively. By applying SDS-PAGE, the molecular weight was confirmed at 35 kDa. The optimal temperature, pH, and isoelectric point were 35 °C, 8.0, 5.6 ± 0.2, respectively. UcB5 was found to have a broad substrate specificity against almost all the tested chromogenic substrates with maximal affinity against N-Succ-Ala-Ala-Pro-Phe-pNA achieving Km of 0.16 mM, Kcat/Km of 3.01 × 105 S−1 M−1, and amidolytic activity of 28.9 µmol min−1 L−1. It was drastically inhibited by TLCK, PMSF, SBTI, and aprotinin while, DTT, β-mercaptoethanol, 2,2′-bipyridine, o-phenanthroline, EDTA, and EGTA had no effect, which suggested a serine protease-type. Also, it has shown a broad substrate specificity against a broad range of natural proteins including serum proteins. A cytotoxicity and electron microscopy study revealed that UcB5 could cause subcellular proteolysis that finally led to liver necrosis. For this, future research should focus on using a combination of external antiproteases and antimicrobial agents for the treatment of microbial diseases instead of using drugs alone.

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Isolation of a putative virulence agent, cytotoxic serine-elastase, from a newly isolated Pseudomonas aeruginosa ZuhP13

Cited by 5 publications

References 21 publications

A survey of elastase-producing bacteria and characteristics of the most potent producer, Priestia megaterium gasm32

A survey of elastase-producing bacteria and characteristics of the most potent producer, Priestia megaterium gasm32

Histological Studies on a Newly Isolated Bacillus subtilis D10 Protease in the Debridement of Burn Wound Eschars Using Mouse Model

A putative cytotoxic serine protease from Salmonella typhimurium UcB5 recovered from undercooked burger

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