Synthesis and assembly of human vault particles in yeast. Issue 12 (25th September 2018)

• Record Type: Journal Article
• Authors: Wang, Meng; Kickhoefer, Valerie A.; Rome, Leonard H.; Foellmer, Oliver K.; Mahendra, Shaily
• Is Part Of: Biotechnology and bioengineering. Volume 115:Issue 12(2018)
• Page Start: 2941
• Page End: 2950
• Publication Date: 2018-09-25
• Subjects: biodegradation -- constitutive expression -- immobilization -- nanoparticle -- peroxidase; Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.26825
• Languages: English
• ISSNs: 0006-3592

Abstract

Abstract: Vault particles are the largest naturally occurring ribonucleoprotein complexes found in the cytoplasm. In all 78 copies of major vault protein (MVP) assemble on polyribosome templates, forming recombinant vault particles, which are of great interest as encapsulation carriers for therapeutics delivery and enzyme stabilization. Baculovirus‐insect cell expression is the only system that has been developed for recombinant vault synthesis, but it has low scalability and slow production rate. In this study, we demonstrated the first use of yeast cells for the production of vault particles with full integrity and functionality solely by expressing the complementary DNA (cDNA) encoding MVP. Vaults synthesized in Pichia pastoris yeast cells are morphologically indistinguishable from recombinant vault particles produced in insect cells, and are able to package and stabilize enzymes resulting in improved longevity and catalytic efficiency. Thus, our results imply that the yeast system is an economical alternative to insect cells for the production of recombinant vaults. The consistency of vault morphology between yeast and insect cell systems also underlines that polyribosome templating may be conserved among eukaryotes, which promises the synthesis and assembly of recombinant human vault particles in other eukaryotic organisms. Abstract : Vault particles are the largest naturally occurring ribonucleoprotein complexes found in cells. Recombinant vault particles, assembled from 78 copies of major vault protein, are of great interest as encapsulation carriers for therapeutics delivery and enzyme stabilization. Here, we demonstrate the use of yeast cells for producing vault particles with full integrity and functionality solely by expressing the MVP cDNA. Vaults synthesized in Pichia pastoris yeast cells are able to package and stabilize enzymes resulting in improved longevity and catalytic efficiency.