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四种乳铁传递蛋白肽在无细胞蛋白质合成系统中的从头表达及抗菌潜力

De novo expression and antibacterial potential of four lactoferricin peptides in cell-free protein synthesis system.

作者信息

El-Baky Nawal Abd, Elkhawaga Maie Ahmed, Abdelkhalek Eman Shawky, Sharaf Mona Mohammed, Redwan Elrashdy Mustafa, Kholef Hoda Reda

机构信息

Therapeutic and Protective Proteins Laboratory, Protein Research Department, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, New Borg EL-Arab, 21934, Alexandria, Egypt.

Botany and Microbiology Department, Faculty of Science, Al-Azhar University, Cairo, Egypt.

出版信息

Biotechnol Rep (Amst). 2020 Dec 26;29:e00583. doi: 10.1016/j.btre.2020.e00583. eCollection 2021 Mar.

Abstract

For the first time, we produced four lactoferricin (LFcin) peptides by a cell-free () method. These short antimicrobial peptides were expressed in an cell-free protein synthesis (CFPS) system and the bioactivity of the produced peptides was demonstrated. Additionally, we designed a novel synthetic consensus peptide (ConLFcin). The genes of bovine Lfcin (bLFcin), human Lfcin (hLFcin), camel Lfcin (cLFcin), and ConLFcin were cloned into pET101/D-TOPO vector then peptides were synthesized by CFPS system. The antibacterial activity of these synthesized peptides was evaluated against , and methicillin-resistant (MRSA). The four cell-free synthesized peptides showed significant antibacterial potency at minimum inhibitory concentration (MIC) values between 1.25 and 10 μg/mL. cLFcin and ConLFcin showed higher antibacterial effects than bLFcin and hLFcin. Thus, cell-free expression system is an ideal system for rapid expression of functionally active short bioactive peptides.

摘要

我们首次通过无细胞方法制备了四种乳铁蛋白肽(LFcin)。这些短抗菌肽在无细胞蛋白质合成(CFPS)系统中表达,并证明了所产生肽的生物活性。此外,我们设计了一种新型合成共有肽(ConLFcin)。将牛Lfcin(bLFcin)、人Lfcin(hLFcin)、骆驼Lfcin(cLFcin)和ConLFcin的基因克隆到pET101/D-TOPO载体中,然后通过CFPS系统合成肽。评估了这些合成肽对大肠杆菌、金黄色葡萄球菌和耐甲氧西林金黄色葡萄球菌(MRSA)的抗菌活性。四种无细胞合成肽在1.25至10μg/mL的最低抑菌浓度(MIC)值下显示出显著的抗菌效力。cLFcin和ConLFcin比bLFcin和hLFcin表现出更高的抗菌效果。因此,无细胞表达系统是快速表达功能活性短生物活性肽的理想系统。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1fac/7779732/a50a660c3dcb/gr1.jpg

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