Efficient production of endotoxin depleted bioactive α-hemolysin of uropathogenic Escherichia coli. Issue 6 (3rd July 2019)

• Record Type: Journal Article
• Authors: Verma, Vivek; Gupta, Surbhi; Kumar, Parveen; Rawat, Ankita; Singh Dhanda, Rakesh; Yadav, Manisha
• Is Part Of: Preparative biochemistry & biotechnology. Volume 49:Issue 6(2019)
• Page Start: 616
• Page End: 622
• Publication Date: 2019-07-03
• Subjects: HlyA -- HlyC -- α-hemolysin -- affinity chromatography -- hemolysis and E. coli; Biochemistry -- Technique -- Periodicals; Biotechnology -- Technique -- Periodicals; Biochemistry -- methods -- Periodicals; Biotechnology -- methods -- Periodicals; 660.6
• DOI: 10.1080/10826068.2019.1591993
• Languages: English
• ISSNs: 1082-6068

Abstract

Abstract: Uropathogenic E. coli (UPEC), especially associated with severe urinary tract infections (UTI) pathologies, harbors an important virulence factor known as α-hemolysin (110 kDa). Hemolytic activity of α-hemolysin (HlyA) requires modification (acylation) of two lysine residues of HlyA by HlyC, part of operon hlyCABD . Most of the previous studies had used whole operon hlyCABD and gene tolC cloning for the production of active α-hemolysin. Studies involving α-hemolysin are limited due to the cumbersome and manual method of purification for this toxin. Here, we report a simple method for production of both active and inactive recombinant α-hemolysin by cloning only hlyA and hlyC genes of operon hlyCABD. Presence of both active and inactive α-hemolysin would be advantageous for functional characterization. After translation, the yield of the purified α-hemolysin was 1 mg/200 ml. Functionality of the recombinant α-hemolysin protein was confirmed using hemolytic assay. This is the first report of the production of active and inactive recombinant α-hemolysin for functional studies.