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Periplasmic production of the recombinant eugenol synthase from Ocimum basilicum | ||
| Trends in Pharmaceutical Sciences | ||
| مقاله 2، دوره 5، شماره 3، آذر 2019، صفحه 131-136 اصل مقاله (487.56 K) | ||
| نوع مقاله: Original Article | ||
| شناسه دیجیتال (DOI): 10.30476/tips.2019.83489.1027 | ||
| نویسندگان | ||
| Neda Mazarei1؛ Anahita Moradi1؛ Seyyed Soheil Rahmatabadi1؛ Shiva Hemmati* 2، 3، 4 | ||
| 1Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran. | ||
| 2Department of Pharmaceutical Biotechnology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran. | ||
| 3Pharmaceutical Sciences Research Center, Shiraz University of Medical Sciences, Shiraz, Iran. | ||
| 4Biotechnology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran. | ||
| چکیده | ||
| Eugenol, the main bioactive compound of clove oil, is a simple molecule with considerable pharmacological properties, including analgesic, antimicrobial, and anti-inflammatory effects. Eugenol is synthesized in several plant species such as Ocimum basilicum (basil) through the activity of eugenol synthase enzyme. Regarding the valuable applications of eugenol as well as notable advantages of the secretory production of recombinant proteins, the excretory production of eugenol synthase has been targeted in this study. Eugenol synthase coding sequence was inserted after pelB peptide in the pET22b vector and expressed in E. coli to investigate if the secretory production of the recombinant protein is possible. Additionally, enzyme purification by Ni-NTA affinity chromatography, extraction of the enzyme from periplasmic fraction, and salting out of the enzyme secreted into the medium by ammonium sulfate precipitation were carried out. Our results showed the presence of eugenol synthase in the cytosol, periplasm, and culture medium with concentrations of about 208.8 µgml-1, 212.5 µgml-1, and 332.0 µgml-1, respectively. The results of this study are useful for further studies to produce large-scale eugenol synthase as an unlimited source of the eugenol compound. | ||
| کلیدواژه‌ها | ||
| Signal peptide؛ Secretory protein؛ Extracellular protein | ||
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