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A reliable approach for pure Yeast Cell Wall protein isolation from Saccharomyces cerevisiae yeast cells

ABSTRACT: Saccharomyces cerevisiae yeast is a fungus presenting a peripheral organelle called the cell wall. The cell wall protects the yeast cell from stress and provides means for communication with the surrounding environment. It has a complex molecular structure, composed of an internal part of cross-linked polysaccharides and an external part of mannoproteins. These latter are very interesting owing to their functional properties, dependent of their molecular features with massive mannosylations. Therefore, the molecular characterization of mannoproteins is a must relying on the optimal isolation and preparation of the cell wall fraction. Multiple methods are well reported for yeast cell wall isolation. The most ap-plied one consists of yeast cell lysis by mechanical disruption. However, applying this classical approach to S288C yeast cells showed a considerable contamination with non-cell wall proteins, mainly comprising mitochondrial proteins. Here-in, we tried to further purify the yeast cell wall preparation by two means: ultracentrifugation and Triton X-100 addition. While the first strategy showed limited outcomes in mitochondrial proteins removal, the second strategy showed optimal results when Triton X-100 was added at 5%, allowing the identification of more mannoproteins and enriching significant-ly their amounts. This promising method could be reliably implemented in lab-scale and industrial processes for “pure” cell wall isolation.

INSTRUMENT(S):

ORGANISM(S): Saccharomyces Cerevisiae (baker's Yeast)

TISSUE(S): Cell Suspension Culture

SUBMITTER: Marie YAMMINE

LAB HEAD: Christian Rolando

PROVIDER: PXD032386 | Pride | 2022-10-15

REPOSITORIES: Pride

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Reliable Approach for Pure Yeast Cell Wall Protein Isolation from <i>Saccharomyces cerevisiae</i> Yeast Cells.

Yammine Marie M Bray Fabrice F Flament Stéphanie S Picavet Antoine A Lacroix Jean-Marie JM Poilpré Emmanuel E Mouly Isabelle I Rolando Christian C

ACS omega 20220815 34

<i>Saccharomyces cerevisiae</i> yeast is a fungus presenting a peripheral organelle called the cell wall. The cell wall protects the yeast cell from stress and provides means for communication with the surrounding environment. It has a complex molecular structure, composed of an internal part of cross-linked polysaccharides and an external part of mannoproteins. These latter are very interesting owing to their functional properties, dependent on their molecular features with massive mannosylatio ...[more]

PMID: 36061670

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			Action	DRS
	BSA.fasta	Fasta
	DP-Replicate1.msf	Msf
	DP-Replicate1.raw	Raw
	DP-Replicate2.msf	Msf
	DP-Replicate2.raw	Raw