A system for the expression and release of heterologous proteins from the core of Bacillus subtilis spores.


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Type
Article
Change log
Authors
Mohamed, Mohamed YH 
Abstract

Methodologies that exploit the durability of Bacillus subtilis spores by displaying heterologous proteins or antigenic molecules on the spore surface for mucosal vaccine delivery and other applications are well established. Here we extend the concept by engineering spores intended as oral delivery vehicles for therapeutic proteins. The method is exemplified by the expression and deposition of human growth hormone in the developing spore core, where the protein is shielded from physicochemical and biological degradation by the protective spore structure. Lysates from physically disrupted spores are shown to stimulate differentiation of a pre-adipocyte cell line to mature adipocyte cells, indicating that the spore-core located human growth hormone is folded correctly and functional. We also introduce a methodology for controlled release of heterologous proteins from the spore core, which utilises components of the PBSX prophage to lyse spores during germination and outgrowth. With further development, spore core expression, coupled with an engineered autolytic germination mechanism, may permit the use of spores as oral delivery carriers of therapeutic proteins.

Description
Keywords
3T3 Cells, Adipocytes, Animals, Bacillus subtilis, Bacteriolysis, Biological Products, Cell Differentiation, Cell Surface Display Techniques, Drug Delivery Systems, Gene Expression, Human Growth Hormone, Humans, Lysogeny, Mice, Prophages, Recombinant Proteins, Spores, Bacterial
Journal Title
FEMS Microbiol Lett
Conference Name
Journal ISSN
0378-1097
1574-6968
Volume Title
365
Publisher
Oxford University Press (OUP)
Sponsorship
MYHM was the recipient of a Yousef Jameel scholarship, administered by the Cambridge Commonwealth, European & International Trust.