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Single-step purification of recombinant anthrax lethal factor from periplasm of Escherichia coli

Paper ID Volume ID Publish Year Pages File Format Full-Text
25679 43590 2006 9 PDF Available
Title
Single-step purification of recombinant anthrax lethal factor from periplasm of Escherichia coli
Abstract

Lethal toxin (LT) that composed by protective antigen and lethal factor (LF) is the major virulence factor of Bacillus anthracis. The treatments of LT in animals could reproduce most manifestations of B. anthracis infections that greatly improves our knowledge in LT-mediated pathogenesis and facilitates anthrax-related researches without having to directly contact the hazardous bacterium B. anthracis. The recombinant protein of LF (rLF), however, still lacks a simple purification method. Herein, we developed single-step nickel affinity purification of rLF with yield up to 3 mg/l. By fusion to the leader sequence of outer membrane protein OmpA, rLF could easily be purified from the periplasm of Escherichia coli. To investigate whether the rLT is functional in our system, both wild type rLF and the catalytic mutant rLF that contains a single amino acid substitution at zinc-binding site (LFE687A), were subjected to macrophage cytotoxicity analysis. Our data showed that the rLT is fully functional, while the LFE687A fail to induce cell death of tested macrophage cells. These findings suggested that the purification protocol herein is a user-friendly method that allows researchers to obtain the functional rLF by single-step purification.

Keywords
Bacillus anthracis; Lethal factor; Recombinant protein; Single-step purification; Periplasm
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Single-step purification of recombinant anthrax lethal factor from periplasm of Escherichia coli
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 126, Issue 3, 10 November 2006, Pages 277–285
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us