Self-assembled amyloid-like oligomeric-cohesin Scaffoldin for augmented protein display on the saccharomyces cerevisiae cell surface.

Applied and Environmental Microbiology
Zhenlin HanWei Wen Su

Abstract

In this study, a molecular self-assembly strategy to develop a novel protein scaffold for amplifying the extent and variety of proteins displayed on the surface of Saccharomyces cerevisiae is presented. The cellulosomal scaffolding protein cohesin and its upstream hydrophilic domain (HD) were genetically fused with the yeast Ure2p N-terminal fibrillogenic domain consisting of residues 1 to 80 (Ure2p(1-80)). The resulting Ure2p(1-80)-HD-cohesin fusion protein was successfully expressed in Escherichia coli to produce self-assembled supramolecular nanofibrils that serve as a novel protein scaffold displaying multiple copies of functional cohesin domains. The amyloid-like property of the nanofibrils was confirmed via thioflavin T staining and atomic force microscopy. These cohesin nanofibrils attached themselves, via a green fluorescent protein (GFP)-dockerin fusion protein, to the cell surface of S. cerevisiae engineered to display a GFP-nanobody. The excess cohesin units on the nanofibrils provide ample sites for binding to dockerin fusion proteins, as exemplified using an mCherry-dockerin fusion protein as well as the Clostridium cellulolyticum CelA endoglucanase. More than a 24-fold increase in mCherry fluorescence and an 8-fol...Continue Reading

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Citations

Oct 26, 2012·Biotechnology and Bioengineering·Zhenlin HanWei Wen Su
Dec 25, 2017·Applied Microbiology and Biotechnology·Zhenlin Han, Wei Wen Su
Jan 4, 2021·Bioprocess and Biosystems Engineering·Mengqi YeGuanjun Chen
Aug 16, 2019·Journal of Biotechnology·Bei ZhangWei Wen Su
Nov 27, 2020·Frontiers in Cellular Neuroscience·Marit A de Beer, Ben N G Giepmans
Mar 1, 2018·ACS Applied Materials & Interfaces·Hyeon-Yeol ChoJeong-Woo Choi
Apr 29, 2015·Industrial & Engineering Chemistry Research·Mason R SmithFei Wen

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