Screening and production of thrombolytic enzymes from different soil environments in Egypt 1
Résumé
Abstract BACKGROUND: Thromboembolic disorders including Myocardial infarction[ MI] and stroke represent the leading causes of death worldwide. Fibrinolytic enzymes extracted from microbial sources have shown high efficacy in dissolving various types of blood clots. AIM OF THE STUDY: Screening and bacterial recombinant DNA manufacturing of bacterial fibrinolytic enzymes from different soil environments in Egypt. METHODOLOGY: A total of 50 soil samples were screened for fibrinolytic enzyme production. The potent fibrinolytic enzyme-producing bacterial isolates were evaluated for their thrombolytic activities using the plasma plate method. The characterization of the enzyme activity and thermostability was carried out; also, the determination of molecular mass using mass spectrometry and western blot technique was performed. The purification of the fibrinolyric enzymes was done through affinity chromatography. On the other hand the production of the purified bacterial fibrinolytic enzyme was achieved via bacterial recombinant DNA technology. Formulation of fibrinolytic enzyme in different dosage forms were tried. RESULTS: In the present study the purified bacterial fibrinolytic enzyme was approximately 83% pure through affinity chromatography and SDS-PAGE assay. The predominating bacterial isolate secreting fibrinolytic enzymes found to be Bacillus cereus. The fibrinolytic enzyme activities of 50 soil samples were found to be in the range of 0.167-1.853 U/ ml. The fibrinolytic activity of extracellular enzyme refined from the culture supernatant of Bacillus cereus was relatively stable over PH range from 7.4-10 for 18 hours and also it showed a stability with highest productivity at a temperature of 43 ̊C for nearly 60 minutes. The enzyme was called fibrin-proteinase[ FP] in the present study. The FP degraded the fibrin clots via direct thrombolysis. The specific activity and the molecular mass of the FP were estimated to be nearly 40.71 Units/ mg protein and nearly 30 KDa respectively. Protease activity was noticed to be slightly increased in presence of Fe+3; While it was inhibited in presence of EDTA. On the other hand, the FP fibrinolytic activity was enhanced greatly after addition of monovalent cations such as K+1 and Na +1. Enzyme was also active over temperature range 20- 65 ̊̊C with the highest activity was at 55 ̊C. The optimum temperature and pH of protease production was at 35-43 ̊̊C and pH 8.0-10. Conclusion: During the present study, the thermostable fibrinolytic enzyme FP purified from Bacillus cereus in different soil environments in Egypt was a promising fibrinolytic enzyme because it showed a broader efficacious fibrinolytic activity as well as less adverse effects than the current fibrinolytic enzymes globally. It is recommended in the future advanced exploration of the optimization of suitable formulation of bacterial fibrinolytic enzymes to aid in the dissolution of various blood clots and as a consequence overcoming the high rate of mortality globally.
Citer ce document
Accès au document
Texte intégral en lecture en ligne, réservé aux abonnés SPHAERO et aux membres de l'institution. Se connecter
Voir l'article sur le site de la revueAuteur(s)
Statistiques
Consultations : 1
Téléchargements : 0