Downloads
Abstract
Listeriolysin O (LLO) is an important extracellular toxin of Listeria monocytogenes which degrades the phospholipid membranes of the host cells’ phagosomes at low pH during the intracellular survival. In contrast to the natural form, the mutant LLO carrying two replacements of amino acid E247M and D320K possesses stable activity at pH 7.4. In this study, we have constructed vectors that carry the mhlyA gene encoding double-mutated LLOE247M/D320K express in B. subtilis 1012. The target gene was fused to the sequence of LysRN-6xHis-TEV site to enhance the recombinant protein expression in B. subtilis and to ease the acquisition of protein by Ni2+-based affinity chromatography. As results, we have obtained the purified protein LLOE247M/D320K with high purity. The activity measurement with 3 HU hemolytic toxins in the pH range from 5.0 to 8.5 suggested that the activity of LLOE247M/D320K was much better than that of natural LLO at neutral pH and slightly alkaline. These results not only provided important scientific foundations for mutant LLO bases expression in B. subtilis but also supplied purified materials for researche and medical applications based on this protein.
Issue: Vol 19 No 3 (2016)
Page No.: 20-31
Published: Sep 30, 2016
Section: Natural Sciences - Research article
DOI: https://doi.org/10.32508/stdj.v19i3.470
Download PDF = 1876 times
Total = 1876 times
Most read articles by the same author(s)
- Thinh Duc Nguyen, Thu Thi Nguyet Nguyen, Diem Ngoc Duong, Son Pham Ngoc Chu, Thuoc Linh Tran, Preparation of an immunoaffinity chromatography column for chloramphenicol binding , Science and Technology Development Journal: Vol 19 No 4 (2016)
- Thinh Duc Nguyen, Thu Thi Nguyet Nguyen, Diem Ngoc Duong, Son Pham Ngoc Chu, Thuoc Linh Tran, Preparation of anti-chloramphenicol rabbit antibody , Science and Technology Development Journal: Vol 19 No 4 (2016)
- Phuong Thi Kim Huynh, Thanh Kieu Huynh, Nam Tri Vo, Anh Le Tuan Nguyen, Hoang Duc Nguyen, Trang Thi Phuong Phan, Thong Van Nguyen, Giang Ha Pham, Establishment of a Multiplex – PCR protocol for detection of Y chromosome microdeletion in Azoospermia male patients , Science and Technology Development Journal: Vol 18 No 4 (2015)