A Comparison of Molecular Investigation of the Gene Encoding the Protease Enzyme Extracted from the Commercially Available and Locally Produced Agaricus Bisporus Fungus

Document Type : New and original researches in the field of Microbiology.

Authors

1 Department of Biology, College of Education for Pure Sciences, Tikrit University, Iraq

2 Enviromental Biotechnology Department, Biotechnology Research Center, Al-Nahrain University, Iraq

Abstract

Background: A protease enzyme extracted from the fungus Agaricus bisporus enhances its use against certain types of cancer, such as breast cancer. Objective : This study compared the molecular investigation of the gene encoding the protease enzyme extracted from both commercially available and locally produced Agaricus bisporus fungus. Methodology: The current study extracted the protease enzyme from the Iraqi Agaricus bisporus fungus, readily available in local markets. The extraction used ethyl alcohol to break down the yeast cell wall. The enzyme was then extracted and purified through several steps outlined in a previous study. The genetic material (DNA) was extracted from Agaricus bisporus using the Promega Genomic DNA Purification Kit to identify the gene encoding the protease enzyme and determine its nucleotide sequences using sequencing technology and the SANGER method. Results: The study results showed that the enzyme's specific activity in the crude extract was 2.67 units/mg. Subsequent purification steps, including precipitation with ammonium sulfate at 80% saturation, raised the specific activity to 5.59 units/mg of protein. The purification process achieved a 2.08-fold increase, with an enzyme yield of 15.81. Conclusion: A. bisporus protease showed increased activity (2.08-fold purification, 5.59 U/mg) with successful gene sequencing, confirming its potential as a biotechnologically promising anticancer agent.

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