Conflict of interest: none to declare.
Tunable nano-oleosomes derived from engineered Yarrowia lipolytica†
Article first published online: 5 NOV 2012
Copyright © 2012 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 110, Issue 3, pages 702–710, March 2013
How to Cite
Han, Z., Madzak, C. and Su, W. W. (2013), Tunable nano-oleosomes derived from engineered Yarrowia lipolytica. Biotechnol. Bioeng., 110: 702–710. doi: 10.1002/bit.24761
- Issue published online: 18 JAN 2013
- Article first published online: 5 NOV 2012
- Accepted manuscript online: 23 OCT 2012 01:25PM EST
- Manuscript Accepted: 11 OCT 2012
- Manuscript Revised: 3 OCT 2012
- Manuscript Received: 10 JUL 2012
- USDA NIFA. Grant Number: 2010-65504-20349
- Hawaii Community Foundation. Grant Numbers: 44272, 11ADVC-49237
- USDA TSTAR Research Program. Grant Number: 2008-34135-19407
- Yarrowia lipolytica;
- surface display;
- self assembly
Oleosomes are discrete organelles filled with neutral lipids surrounded by a protein-embedded phospholipid monolayer. Their simple yet robust structure, as well as their amenability to biological, chemical, and physical processing, can be exploited for various biotechnology applications. In this study, we report facile biosynthesis of functionalized oleosomes within oleaginous yeast Yarrowia lipolytica, through expression of oleosin fusion proteins. By fusing a cDNA clone of a sesame oleosin with either the coding sequence of a red fluorescent protein mCherry or a cellulosomal scaffolding protein cohesin from Clostridium cellulolyticum, these oleosin-fusion proteins were efficiently expressed and specifically targeted to and anchored on the surface of the oleosomes within the Y. lipolytica cells. The engineered oleosomes can be easily separated from the Y. lipolytica cell extract via floating centrifugation and both mCherry and cohesin domains are shown to be functional. Upon sonication, the engineered Yarrowia oleosomes exhibit a mean diameter of 200–300 nm and are found to be highly stable. The feasibility of co-displaying multiple proteins on the Yarrowia oleosomes was demonstrated by incubating cohesin-displaying oleosomes with different dockerin-fusion proteins. Based on this strategy, engineered oleosomes with both cell-targeting and reporting activities were created and shown to be functional. Taken together, the Yarrowia oleosome surface display system in which oleosin serves as an efficient membrane anchor motif shows great promise as a simple platform for creating tunable nanoparticles. Biotechnol. Bioeng. 2013; 110: 702–710. © 2012 Wiley Periodicals, Inc.