Structural study and sub-cloning of a type of endolysin

  • سال انتشار: 1399
  • محل انتشار: بیست و یکمین کنگره ملی و نهمین کنگره بین المللی زیست شناسی ایران
  • کد COI اختصاصی: BIOCONF21_0726
  • زبان مقاله: انگلیسی
  • تعداد مشاهده: 198
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نویسندگان

Sepideh Yazdianpour

Faculty of biotechnology, campus of modern Science and technology, Semnan university

Shamsozoha Abolmaali

Faculty of biology, of basic sciences, Semnan University

Shakiba Darvishalipour astaneh

Faculty of biotechnology, campus of modern Science and technology, Semnan university

چکیده

Identification of new antibacterial compounds as a treatment is a way to solve the health care against antibiotic resistant infections. Lysines, are one of antibacterial agent against to remove antibiotic resistant infections.N-acetylmuramyl-L-alanine amidases, also known as Endolysins, which encode in phage’s genome are ableto break down the amide bonds in the peptidoglycan wall of bacterial. The physicochemical properties and secondary and tertiary structure of this enzyme were studied by bioinformatics softwares and the predicted- tertiary structures evaluated by q-mean software (۰.۶% confidence). The synthetic gene encoding N-acetyl muramyl-L-alanine amidase of SPP۱ phage was subcloned in pET۲۸a vector. The clone was confirmed by enzymatic digestion.The exoression of the recombinant protein was induced by IPTG ۱۰۰mM and purified by ۱۰۰mM imidazole gradient using a Nickel column chromatography. Toremove salts and imidazole, recombinant enzyme was dialyzed against PBS buffer for ۳ h. Bioinformatics studies showed that this protein has pI ۹.۶, molecular weight ۳۰ kDa, alpha index ۸۳.۸۷ and a polarity index of -۰.۴۹. The secondary structure of this protein includes of ۳۴.۳۲% of alpha helix, ۶.۶۴% of beta sheets, ۲۳% of extended strands, and ۳۵% of random coils. The ۳۳kDa band enzyme observed in SDS-PAGE ۱۲%. Surveying the enzymatic reactions for the antibacterial effect on E.coli and S. aureus is under processing

کلیدواژه ها

Endolysin,Phage SSP۱, N-acetylmuramyl-L-alanine amidases, recombinant Gene

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