fa بررسی بیان آنزیم رامنوزیل ‌ترانسفراز در مسیر تولید بیوسورفکتانت باکتری سودوموناس آئروژینوزا درحضور نانوساختار آهن برپایه SDS میکروبیولوژی Microbiology مقاله پژوهشی Research Article <span style="font-size:10pt"><span style="background:white"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span lang="AR-SA" style="font-size:16.0pt"><span b="" nazanin="" style="font-family:">سابقه و هدف:</span></span></b> <span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">رامنولیپیدها از</span></span> <span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">مهم&shy;ترین</span></span> <span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">بیوسورفکتانت&shy;های گلیکولیپیدی تولید شده توسط باکتری <i>سودوموناس&shy;آئروژینوزا</i></span></span><i> </i><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">هستند. در این مطالعه بیان پروتئین&shy;های تولیدکننده بیوسورفکتانت باکتری <i>سودوموناس آئروژینوزا</i> در حضور نانوساختار</span></span> <span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">آهن با پوشش سدیم&shy;دودسیل سولفات (</span></span><span dir="LTR" style="font-size:11.0pt">Fe/SDS</span><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">) مورد ارزیابی قرار گرفت.</span></span></span></span></span></span></span><br> <span style="font-size:10pt"><span style="background:white"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span lang="AR-SA" style="font-size:16.0pt"><span b="" nazanin="" style="font-family:">مواد و روش &shy;ها</span></span></b><b><span dir="LTR" style="font-size:11.0pt">:</span></b> <span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">بیان آنزیم <i>رامنوزیل </i></span></span><i><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">ترانسفراز</span></span></i><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> (</span></span><span dir="LTR" style="font-size:11.0pt">Surfactant Protein A</span><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">) توسط</span></span><span dir="LTR" style="font-size:11.0pt">SDS-PAGE </span><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">&nbsp;و وسترن&shy;بلات بررسی گردید. آنزیم <i>رامنوزیل ترانسفراز</i>، روی ژل</span></span><span dir="LTR" style="font-size:11.0pt"> SDS-PAGE </span><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">آنالیز شد و توسط وسترن&shy;بلات تأیید گردید. وجود نانوذرات</span></span> <span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">برسطح باکتری توسط </span></span>&nbsp;<span dir="LTR" style="font-size:11.0pt">TEM</span><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">و مورفولوژی نانوذرات توسط </span></span><span dir="LTR" style="font-size:11.0pt">SEM</span><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> بررسی گردید. </span></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:normal"><span style="tab-stops:131.05pt"><span style="direction:rtl"><span style="unicode-bidi:embed"><span calibri="" style="font-family:"><b><span lang="AR-SA" style="font-size:16.0pt"><span b="" nazanin="" style="font-family:">یافته &shy;ها</span></span></b><b>:</b> <span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">مطابق </span></span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span dir="LTR" style="font-size:10.0pt">SEM</span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">، اندازه نانوذرات</span></span></span></span> <span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">20 نانومتر و کروی شکل دیده شد. اتصال نانوذرات </span></span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span dir="LTR" style="font-size:10.0pt">Fe/SDS</span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> به بیوسورفکتانت &shy;توسط </span></span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span dir="LTR" style="font-size:10.0pt">TEM</span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> تأیید شد. آنزیم رامنوزیل ترانسفراز با وزن مولکولی40 کیلودالتون در نمونه آزمون بیان شد. بررسی</span></span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> نتایج بیان پروتئین نشان داد، </span></span></span></span><span b="" nazanin="" style="font-family:">حضور</span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> آنزیم رامنوزیل ترانسفراز تحت تأثیر نانوذرات </span></span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span dir="LTR" style="font-size:10.0pt">Fe/SDS</span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> افزایش پیدا کرد و نسبت&shy;به نانوذرات دیگر و نمونه کنترل بالاترین</span></span></span></span><span b="" nazanin="" style="font-family:"> حضور</span> <span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">را نشان داد، درحالی&shy;که نانوذرات </span></span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span dir="LTR" style="font-size:10.0pt">SDS</span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> و </span></span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span dir="LTR" style="font-size:10.0pt">Fe</span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> به&shy;تنهایی در تأیید پروتئین</span></span></span></span> <span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">مذکور</span></span></span></span> <span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">مؤثر واقع نشده و کمابیش مشابه نمونه کنترل است. نتایج وسترن</span></span></span></span> <span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">بلاتینگ </span></span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span dir="LTR" style="font-size:10.0pt">SDS-PAGE</span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> تأییدی بر آنزیم رامنوزیل ترانسفراز تحت تأثیر نانوذرات </span></span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span dir="LTR" style="font-size:10.0pt">Fe/SDS</span></span></span><span class="Char" style="background:white"><span new="" roman="" style="font-family:" times=""><span style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> بود.</span></span></span></span> <b><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"><span style="color:black"></span></span></span></b></span></span></span></span></span></span><br> <span style="font-size:10pt"><span style="background:white"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><b><span lang="AR-SA" style="font-size:16.0pt"><span b="" nazanin="" style="font-family:">نتیجه &shy;گیری</span></span></b><b><span dir="LTR" style="font-size:11.0pt">:</span></b> <span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">نتایج پژوهش&shy; حاضر بیانگر </span></span><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">اهمیت نانوذرات </span></span><span dir="LTR" style="font-size:11.0pt">Fe/SDS</span><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> در تأیید </span></span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">آنزیم <i>رامنوزیل&shy;ترانسفراز</i></span></span><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:"> در مسیر تولید </span></span><span lang="AR-SA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">بیوسورفکتانت از <i>سودوموناس آئروژینوزا </i></span></span><span lang="FA" style="font-size:12.0pt"><span b="" nazanin="" style="font-family:">است.</span></span></span></span></span></span></span><br> <span style="font-size:10pt"><span style="background:white"><span style="direction:rtl"><span style="unicode-bidi:embed"><span new="" roman="" style="font-family:" times=""><span lang="AR-SA" style="font-size:12.0pt"><span style="font-family:"B Nazanin""></span></span></span></span></span></span></span> <span style="font-size:11pt"><span style="background:white"><span style="line-height:normal"><span style="tab-stops:45.8pt 91.6pt 137.4pt 183.2pt 229.0pt 274.8pt 320.6pt 366.4pt 412.2pt 458.0pt 503.8pt 549.6pt 595.4pt 641.2pt 687.0pt 732.8pt"><span calibri="" style="font-family:"><b><span new="" roman="" style="font-family:" times="">Aim and Background</span></b><span new="" roman="" style="font-family:" times="">:</span> <i><span style="background:white"><span new="" roman="" style="font-family:" times="">Rhamnolipids</span></span></i><span style="background:white"><span new="" roman="" style="font-family:" times=""> are one of the most important bio surfactants of glycolipids produced by Pseudomonas aeruginosa. In this study, the expression of bio surfactant producing proteins of Pseudomonas aeruginosa in the presence of sodium dodecyl sulfate coated iron nanostructure (Fe/SDS) were evaluated. </span></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="background:white"><span style="line-height:normal"><span style="tab-stops:45.8pt 91.6pt 137.4pt 183.2pt 229.0pt 274.8pt 320.6pt 366.4pt 412.2pt 458.0pt 503.8pt 549.6pt 595.4pt 641.2pt 687.0pt 732.8pt"><span calibri="" style="font-family:"><b><span new="" roman="" style="font-family:" times="">Materials and Methods:</span></b> <span style="background:white"><span new="" roman="" style="font-family:" times="">The expression of Surfactant Protein A enzyme was investigated by SDS-PAGE and Western blot. The enzyme rhamnosyl transferase was analyzed on SDS-PAGE gel and confirmed by Western blotting. The presence of nanoparticles on the surface of bacteria and the morphology of nanoparticles were investigated by TEM and SEM. </span></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="background:white"><span style="line-height:normal"><span style="tab-stops:45.8pt 91.6pt 137.4pt 183.2pt 229.0pt 274.8pt 320.6pt 366.4pt 412.2pt 458.0pt 503.8pt 549.6pt 595.4pt 641.2pt 687.0pt 732.8pt"><span calibri="" style="font-family:"><b><span new="" roman="" style="font-family:" times="">Results:</span></b><span new="" roman="" style="font-family:" times=""> According to SEM, nanoparticles with a size of 20 nm and a spherical shape were seen. </span><span style="background:white"><span new="" roman="" style="font-family:" times="">Binding of Fe / SDS nanoparticles to bio surfactant was confirmed by TEM. The enzyme rhamnosyl transferase with a molecular weight of 40 kDa was expressed in the test sample. The results of protein expression showed that the confirmed of rhamnosyl transferase enzyme increased significantly under the influence of Fe/SDS nanoparticles and showed the confirmed compared to other nanoparticles and the control sample. While SDS and Fe nanoparticles alone are not effective in confirmed this protein and are almost identical to the control sample. Western blotting results of SDS-PAGE confirmed the expression of <i>rhamnolipid</i> transferase under the influence of Fe/SDS nanoparticles. </span></span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="background:white"><span style="line-height:normal"><span style="tab-stops:45.8pt 91.6pt 137.4pt 183.2pt 229.0pt 274.8pt 320.6pt 366.4pt 412.2pt 458.0pt 503.8pt 549.6pt 595.4pt 641.2pt 687.0pt 732.8pt"><span calibri="" style="font-family:"><b><span new="" roman="" style="font-family:" times="">Conclusion:</span></b><span style="background:white"><span new="" roman="" style="font-family:" times=""> The results of the present study indicate the importance of Fe/SDS nanoparticles in increasing the expression of <i>rhamnolipid</i> transferase enzyme in the production of bio surfactants from Pseudomonas aeruginosa.</span></span></span></span></span></span></span><br> &nbsp; بیوسورفکتانت, Fe/SDS, سودوموناس آئروژینوزا, وسترن بلات, SDS-PAGE, Iau Science Bio surfactant, Fe/SDS nanoparticle, Pseudomonas aeruginosa, Western blot, SDS-PAGE, Iau Science 35 44 http://ncmbjpiau.ir/browse.php?a_code=A-10-1788-1&slc_lang=fa&sid=1 Yaser Ahsani Arani یاسر احسنی ارانی ahsani.yaser@gmail.com 100319475328460013653 100319475328460013653 No Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran گروه زیست شناسی، واحد علوم و تحقیقات تهران، دانشگاه آزاد اسلامی، تهران، ایران Zahra Noormohammadi زهرا نورمحمدی z-nouri@srbiau.ac.ir 100319475328460013654 100319475328460013654 No Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran گروه زیست شناسی، واحد علوم و تحقیقات تهران، دانشگاه آزاد اسلامی، تهران، ایران Behnam Rasekh بهنام راسخ rasekhb@ripi.ir 100319475328460013655 100319475328460013655 Yes Director Environment & Biotechnology Research Division, Research Institute of Petroleum Industry (RIPI), Tehran, Iran گروه پژوهش میکروبیولوژی و بیوتکنولوژی، پژوهشگاه صنعت نفت تهران، تهران، ایران Fatemeh Yazdian فاطمه یزدیان yazdian@ut.ac.ir 100319475328460013656 100319475328460013656 No Department of Life Science Engineering, Faculty of New Science and Technologies, University of Tehran, Tehran, Iran گروه مهندسی علوم زیستی، دانشکده علوم و فنون نوین، دانشگاه تهران، تهران، ایران Hojjat Kazemi حجت کاظمی Hojjatkazemi4@yahoo.com 100319475328460013657 100319475328460013657 No Analytical Chemistry Research Group, Research Institute of Petroleum Industry (RIPI), Tehran, Iran گروه پژوهش شیمی تجزیه، پژوهشگاه صنعت نفت تهران، تهران، ایران