Optimization of ammonium sulfate precipitation of a chimeric peptidoglycan hidrolysing Enzme
Publish place: Twenty-second Iranian Congress of Microbiology (Virtual)
Publish Year: 1400
نوع سند: مقاله کنفرانسی
زبان: English
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شناسه ملی سند علمی:
MEDISM22_275
تاریخ نمایه سازی: 8 مهر 1400
Abstract:
Background and Aim : Endolysins are essential bacteriophage encoded enzymes in the lytic phage life cycle, which hydrolyze the host cell wall and are a good antibiotic substituent against Antibacterial resistance. The endolysins of Gram-positive bacteria infecting bacteriophages contain catalytic domain(s) and binding domain. Histidine-dependent Amidohydrolase Peptidase (CHAP) domain is one of the most active domains, which has Anti-Staphylococcus aureus activity.Methods : The chimeric protein that contains the CHAP domain and binding domain was expressed in BL۲۱(Gold). Bacterial cells were sonicated and the chimeric protein ammonium sulfate precipitation was optimized. Results : We confirmed the level of precipitated chimeric protein using SDS-PAGE. The highest level of chimeric protein precipitate was observed in ۱۰% ammonium sulfate concentration.Conclusion : In this study, ammonium sulfate precipitation of a new chimeric peptidoglycan hydrolase was done to prepare crude lysate for the next step of purification. Ammonium sulfate precipitation is a useful technique as the first step in protein purification because you can have quick, bulk precipitation of cellular proteins.
Keywords:
Histidine dependent Amidohydrolase Peptidase (CHAP) , Chimeric protein , Ammonium sulfate precipitation , Endolysin
Authors
Yasaman Ahmadbeigi
Department of microbiology and microbial biotechnology, Faculty of life sciences and biotechnology, Shahid Beheshti University, Tehran, Iran.
Neda soleimani
Department of microbiology and microbial biotechnology, Faculty of life sciences and biotechnology, Shahid Beheshti University, Tehran, Iran.
Farzaneh Azizmohseni
Department of Biotechnology, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran
Zahra Amini-bayat
Department of Biotechnology, Iranian Research Organization for Science and Technology (IROST), Tehran, Iran