Flocculation ofSaccharomyces cerevisiae is induced by transformation with theGAP1 gene fromKluyveromyces marxianus
✍ Scribed by Moreira, R. Falc�o; Ferreira-da-Silva, F.; Fernandes, P. A.; Moradas-Ferreira, P.
- Book ID
- 101224496
- Publisher
- John Wiley and Sons
- Year
- 2000
- Tongue
- English
- Weight
- 133 KB
- Volume
- 16
- Category
- Article
- ISSN
- 0749-503X
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✦ Synopsis
A non-¯occulent strain of Saccharomyces cerevisiae was transformed with the GAP1 gene which encodes p37, a GAPDH-like protein present in the cell wall of Kluyveromyces marxianus ¯occulent cells. The transformed cells were characterized with respect to ¯occulation behaviour, morphology, growth, cell wall integrity and GAPDH activity. A ¯occulent phenotype was acquired by the transformed cells, showing a behaviour in respect to ¯occulation/de¯occulation very similar to that of K. marxianus. The presence of p37 in the cell wall was assessed by immunoprecipitation of biotinylated cell wall proteins and an accumulation of p37 was evident in the cell wall of transformed cells. This result was con®rmed by studies using a chimeric protein resulting from fusing the p37 with a yeast-enhanced green ¯uorescent protein, yEGFP. The recombinant protein was localized mainly in the cell wall of the transformed strain, although the presence of p37 in the cytosol was indicated by an increase in GAPDH activity. Calco¯uor white sensitivity tests indicated that the cell wall structure is affected by the accumulation of p37. These results provided further evidence of p37 function regarding ¯occulation and that although lacking a N-terminal signal peptide p37 is targeted to the cell wall.