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Research Journal of Pharmacy and Technology
Year : 2022, Volume : 15, Issue : 12
First page : ( 5415) Last page : ( 5420)
Print ISSN : 0974-3618. Online ISSN : 0974-360X.
Article DOI : 10.52711/0974-360X.2022.00912

Biological Study of protease produced by clinical isolates of Staphylococcus aureus

Abdulrazaq Raghad Abdulatif1, Mahmood Waqas Saadi2, Alwan Baydaa3, Saleh Tahreer Hadi4, Hashim Saba T.5, Al-Rubaii Bahaa Abdullah Laftaah6,*

1Department of Biology, College of Science, Mustansiriyah University, Baghdad, Iraq.

2Tikrit University, College of Science, Biology Department.

3Department of Biology, College of Science, Mustansiriyah University, Baghdad, Iraq.

4Department of Biology, College of Science, Mustansiriyah University, Baghdad, Iraq.

5Department of Biology, College of Science, Mustansiriyah University, Baghdad, Iraq.

6Department of Biology, College of Science, University of Baghdad, Baghdad, Iraq.

*Corresponding Author E-mail: bahaa974@gmail.com, bahaa.abdullah@sc.uobaghdad.edu.iq

Online Published on 06 March, 2023.

Abstract

The bacterial isolates were obtained from Al-Kindi Hospital were diagnosed by the Vitek-2 system and re confirm by 16srRNA gene as S. aurous, the results were shown 20 isolates (66.7%) out of 30 isolates were positive to protease production. All bacterial isolates (100%) were sensitive to Gentamicin and Levofloxacin. but resistant (100%) to aztreonam. The best temperature for enzyme production from bacteria was 37 °C, and the best pH for enzyme production was 7. Partial purification of the bacterial enzyme (protease) was carried out using short steps included ammonium sulfate 65% saturation, ion exchange using DEAE- cellulose column and then applied on gel filtration chromatography using Sephadex G-200 column. The enzymatic activity was determined for each purification step. The specific fold and yield of the purified enzyme were 5.91 and 38.3 % respectively. The molecular weight of the purified enzyme was 37 kDa , it was determined by SDS-PAGE. After being exposed to high concentrations of the protease enzyme (800-1000 µg/ml), the proliferation of a breast cancer cell line (MCF-7) was seen to be suppressed, however the inhibitory effect gradually diminished as the concentration of the enzyme decreased. 200–400 µg/ml is the recommended concentration.

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Keywords

Biological study, Staphylococcus aureus, Protease, Purification, Anticancer.

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