Yeast-Expressed Bacteriophage-Like Particles for the Packaging of Nanomaterials

• Published in: Molecular biotechnology Vol. 56; no. 2; pp. 102 - 110
• Main Authors: Freivalds, Janis, Kotelovica, Svetlana, Voronkova, Tatyana, Ose, Velta, Tars, Kaspars, Kazaks, Andris
• Format: Journal Article
• Language: English
• Published: Boston Springer-Verlag 01.02.2014; Springer US; Springer Nature B.V
• Subjects: Acinetobacter; Amino Acid Sequence; Antiporters - metabolism; Atoms & subatomic particles; Bacteria; Bacteriophages - physiology; Biochemistry; Biological Techniques; Biotechnology; calcium; Capsid Proteins - metabolism; Caulobacter; Cell Biology; Chemistry; Chemistry and Materials Science; coat proteins; coliphages; DNA; Gene expression; gold; Human Genetics; ions; Nanomaterials; nanoparticles; Nanoparticles - chemistry; Nanotechnology; Nucleic acids; Packaging; Peptides; Phage hcb5; Phage pp7; Pichia - classification; Pichia - virology; Pichia pastoris; Protein Science; Pseudomonas; Ribonucleic acid; RNA; RNA, Viral - genetics; Saccharomyces cerevisiae; Saccharomyces cerevisiae - virology; structural genes; vaccine development; Vaccines, Virus-Like Particle - metabolism; Virus Assembly; virus-like particles; Yeast; Yeasts; Packaging; RNA bacteriophages; Nanomaterials; Yeast; Reassembly; Virus-like particles
• ISSN: 1073-6085; 1559-0305
• DOI: 10.1007/s12033-013-9686-0

Virus-like particles (VLPs) generated by heterologous expression of viral structural genes have become powerful tools in vaccine development. Recently, we and others have reported on the assembly of VLPs of the RNA bacteriophages MS2, Qβ, and GA in yeast. Here, we investigate the formation of VLPs of five additional phages in the yeasts Saccharomyces cerevisiae and Pichia pastoris, namely, the coliphages SP and fr, Acinetobacter phage AP205, Pseudomonas phage PP7, and Caulobacter phage φCb5. In all cases except SP, particle formation was detected, although VLP outcome varied from 0.2 to 8 mg from 1 g of wet cells. We have found that phage φCb5 VLPs easily dissociate into coat protein dimers when applied to strong anion exchangers. Upon salt removal and the addition of nucleic acid or its mimics and calcium ions, the dimers re-assemble into VLPs with high efficiency. A variety of compounds, including RNA, DNA, and gold nanoparticles can be packaged inside φCb5 VLPs. The ease with which phage φCb5 coat protein dimers can be purified in high quantities and re-assembled into VLPs makes them attractive for downstream applications including the internal packaging of nanomaterials and the chemical coupling of peptides of interest on the surface.