چاپ صفحه |  صفحه نخست سامانه |  نویسندگان |  اطلاعات تفضیلی |  دانلود مقاله | دانشگاه علوم پزشکی تبریز |
| نویسنده ثبت کننده مقاله | اعظم صفری |
| مرحله جاری مقاله | تایید نهایی |
| دانشکده/مرکز مربوطه | بیماری های بافت همبند |
| کد مقاله | 71878 |
| عنوان فارسی مقاله | Molecular cloning and soluble overexpression of recombinant glutaminase for ALL treatment |
| عنوان لاتین مقاله | Molecular cloning and soluble overexpression of recombinant glutaminase for ALL treatment |
| نوع ارائه | سخنرانی |
| عنوان کنگره / همایش | سومین کنگره بین المللی داروسازی نوین |
| نوع کنگره / همایش | بین المللی |
| کشور محل برگزاری کنگره/ همایش | Iran (Islamic Republic) |
| شهر محل برگزاری کنگره/ همایش | Tehran |
| سال انتشار/ ارائه شمسی | 1398 |
| سال انتشار/ارائه میلادی | 2020 |
| تاریخ شمسی شروع و خاتمه کنگره/همایش | 1398/11/16 الی 1398/11/18 |
| آدرس لینک مقاله/ همایش در شبکه اینترنت | https://www.pharmacyupdates.com/images/pdf/book98.pdf |
| آدرس علمی (Affiliation) نویسنده متقاضی | Connective Tissue Diseases Research Center, Tabriz University of Medical Sciences, Tabriz, Iran |
| نویسنده | نفر چندم مقاله |
|---|---|
| شایان سیمای | اول |
| اعظم صفری | دوم |
| ژاله برار | سوم |
| یداله امیدی | چهارم |
| عنوان | متن |
|---|---|
| خلاصه مقاله | Background: Due to the enormous burden of cancer on society worldwide, the development of novel therapeutic agents and strategies against cancer is one of the popular medical research fields. L-glutaminase (EC 3.5.1.2) is a member of the beta-lactamase superfamily that catalyzes the hydrolytic degradation of L-glutamine to L-glutamic acid. The depletion of the glutamine can starve the tumor cells and lead to activating apoptosis pathways, regulating proliferation rate, and stopping tumor growth. L-glutaminase is widely distributed among different microorganisms. The aims of this study are the identification of the L-glutaminase gene from a new halo-thermotolerant Bacillus, molecular cloning, and optimization of soluble overexpression in prokaryotic expression systems. Methods: In this study, the glutaminase gene (GlsA) from locally isolated Bacillus licheniformis SL-1 was identified and cloned into the pET22b+ expression vector. Recombinant glutaminase was overexpressed in modified Escherichia coli strains, Origami B and BL21. Enzyme production was optimized in different temperatures and IPTG concentrations in both expression systems. Then, extraction was conducted at 4˚C in a protease inhibitor-containing lysis buffer using sonication and freeze-thawing methods. The crude extracts from bacterial cells and expression efficacy were analyzed on 12% SDS-PAGE. The recombinant glutaminase was tagged with a polyhistidine tag at C-terminus and could be efficiently purified by nickel-sepharose beads using immobilized metal affinity chromatography (IMAC) method to apparent homogeneity. Results: From the results, the recombinant glutaminase was significantly overexpressed in the soluble fraction obtained from E. coli BL 21. The yield of the enzyme in E. coli BL21 showed significant improvement over the glutaminase produced in the Origami expression system. From SDS-PAGE analysis, the molecular weight of glutaminase monomers was detected around ∼39 kDa. The optimal condition for recombinant enzyme production was adjusted at 20 °C, 180 rpm, 1 mM IPTG, and OD: 0.7-0.9. Conclusion: The identified glutaminase from new halo-thermotolerant bacillus with high overexpression capacity in prokaryotic systems can be considered as a potential anti-cancer agent in ALL treatment. |
| کلمات کلیدی |
| نام فایل | تاریخ درج فایل | اندازه فایل | دانلود |
|---|---|---|---|
| Abstract.Glu..jpg | 1399/02/01 | 810721 | دانلود |
| R1.Abstract-final.pdf | 1399/02/01 | 231046 | دانلود |
