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Please use this identifier to cite or link to this item: http://hdl.handle.net/10204/5519

Title: Extracellular secretion of a recombinant therapeutic peptide by Bacillus halodurans utilizing a modified flagellin type III secretion system
Authors: Berger, E
Crampton, MC
Nxumalo, NP
Louw, ME
Keywords: Bacillus halodurans
Microbial cells
Issue Date: Aug-2011
Publisher: Biomed Central
Citation: Berger, E, Crampton, MC, Nxumalo, NP et al. 2011. Extracellular secretion of a recombinant therapeutic peptide by Bacillus halodurans utilizing a modified flagellin type III secretion system. Microbial Cell Factories, Vol. 10(62), pp 1-10
Series/Report no.: Workflow request;7183
Abstract: Through modification of the flagellin type III secretion pathway of Bacillus halodurans heterologous peptides could be secreted into the medium as flagellin fusion monomers. The stability of the secreted monomers was significantly enhanced through gene-targeted inactivation of host cell extracellular proteases. In evaluating the biotechnological potential of this extracellular secretion system an anti-viral therapeutic peptide, Enfuvirtide, was chosen. Currently, Enfuvirtide is synthesised utilizing 106 chemical steps. The authors used Enfuvirtide as a model system in an effort to develop a more cost-effective biological process for therapeutic peptide production. Results: An attempt was made to increase the levels of the fusion peptide by two strategies, namely strain improvement through gene-targeted knock-outs, as well as vector and cassette optimization. Both approaches proved to be successful. Through chromosomal inactivation of the spo0A, lytC and lytE genes, giving rise to strain B. halodurans BhFDL05S, the secretion of recombinant peptide fusions was increased 10-fold. Cassette optimization, incorporating an expression vector pNW33N and the N- and C-terminal regions of the flagellin monomer as an inframe peptide fusion, resulted in a further 3.5-fold increase in the secretion of recombinant peptide fusions. Conclusions: The type III flagellar secretion system of B. halodurans has been shown to successfully secrete a therapeutic peptide as a heterologous flagellin fusion. Improvements to both the strain and expression cassette led to increased levels of recombinant peptide, showing promise for a biotechnological application.
Description: Copyright: 2011 Berger et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
URI: http://www.microbialcellfactories.com/content/10/1/62
ISSN: 1475-2859
Appears in Collections:Microbial expression systems
General science, engineering & technology

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