- ISSN: 1999-7086
- E-ISSN: 1999-7094
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oa Production and purification of protease produced by UV-mutated Pseudomonas aeruginosa
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المؤلفون:
Hussein A. Hussein1, Bushra H. Saleh1 and Nedhaal S. Zbar1
- Source: Journal of Emergency Medicine, Trauma and Acute Care, Volume 2024, Issue 5 - The 12th international scientific conference of Al-Nahrain University, نوفمبر ٢٠٢٤, 5
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- ٠٢ مايو ٢٠٢٤
- ٢٧ يونيو ٢٠٢٤
- ١٣ نوفمبر ٢٠٢٤
ملخص
Background: The enzyme protease accounts for approximately two-thirds to one-third of all enzymes used in different industries. Protease is produced in large quantities as a crude enzyme for food industries.
Objectives: The aim of this study was to investigate the effect of UV radiation on the protease produced by Pseudomonas aeruginosa.
Methods: Of a total of 149 samples collected from various clinical cases (wounds, burns, otitis media, and urinary tract infections) in four hospitals in Baghdad for the isolation of P. aeruginosa, 90 isolates were identified as belonging to P. aeruginosa after isolation on a specific selective media, and the Vitek 2 system was later used for final identification. Thirty-six P. aeruginosa isolates were screened for protease production on a skim milk medium.
Results: This study found that most of the P. aeruginosa isolates were protease producers and the isolate p3 was the best, which had a hydrolysis zone of up to 29 mm and their specific activity was 0.134 units (U)/mg protein in the culture filtrate. Therefore, this isolate was selected to study the effect of UV light on the production of the protease enzyme. The results showed that protease was produced at high levels from mutant p3 isolates after exposure to UV irradiation for different durations (15, 25, 35, 45, 55, 65, 75, 85, and 95 seconds). The specific activity of the enzyme in a crude filtrate was 0.65 U/mg protein compared with the wild type (0.134 U/mg protein). Protease produced from the mutated isolate p3 was later purified in three purification steps: first, its precipitate with 70% ammonium sulfate, second through a DEAE-cellulose column, the specific activity of which reached 7 U/mg protein, and in the final step through a Sephadex G-200 column. The purified enzyme protease had a specific activity that gradually increased to 10.5 U/mg with an 18.23-fold purification and 84.1% enzyme recovery.
Conclusion: UV radiation was an efficient method to induce the production of protease enzymes.
