Details
| Original language | English |
|---|---|
| Pages (from-to) | 140-147 |
| Number of pages | 8 |
| Journal | Cytometry Part A |
| Volume | 75 |
| Issue number | 2 |
| Publication status | Published - 12 Dec 2008 |
Abstract
The ability of brewing yeast to flocculate is an important feature for brewing of qualitatively good beer. Flocculation involves two main cell wall structures, which are the flocculation proteins (flocculins) and mannans, to which these flocculins bind. Unfortunately, in practice, the flocculation ability may get lost after several repitches. Flow cytometry was employed to analyze glucose and mannose structures of the cell surface by application of fluorescent lectins. Validation of the expression of the flocculin genes Lg-FLOl, FLO1, FLO5, and FLO9 was carried out using microarray techniques. SDS-PAGE, western blot, and ESI-MS/MS analyses served to isolate and determine yeast cell flocculins. Mannose and glucose labeling with fluorescent lectins allowed differentiating powdery and flocculent yeast cells under laboratory conditions. Using microarray techniques and proteomics, the four flocculation genes Lg-FLOl, FLO1, FLO5, FLO9, and the protein Lg-Flolp were identified as factors of major importance for flocculation. The expression of the genes was several times higher in flocculent yeast cells than in powdery ones. Flow cytometry is a fast and simple method to quantify the proportions of powdery and flocculent yeast cells in suspensions under defined cultivation conditions. However, differentiation under industrial conditions will require mRNA and protein expression profiling.
Keywords
- Flocculation, Lg-flolp, Microarray, Microbial flow cytometry, Proteome, Saccharomyces cerevisiae
ASJC Scopus subject areas
- Medicine(all)
- Pathology and Forensic Medicine
- Medicine(all)
- Histology
- Biochemistry, Genetics and Molecular Biology(all)
- Cell Biology
Cite this
- Standard
- Harvard
- Apa
- Vancouver
- BibTeX
- RIS
In: Cytometry Part A, Vol. 75, No. 2, 12.12.2008, p. 140-147.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Prediction of flocculation ability of brewing yeast inoculates by flow cytometry, proteome analysis, and mRNA profiling
AU - Heine, Franziska
AU - Stahl, Frank
AU - Sträuber, Heike
AU - Wiacek, Claudia
AU - Benndorf, Dirk
AU - Repenning, Cornelia
AU - Schmidt, Frank
AU - Scheper, Thomas
AU - Von Bergen, Martin
AU - Harms, Hauke
AU - Müller, Susann
PY - 2008/12/12
Y1 - 2008/12/12
N2 - The ability of brewing yeast to flocculate is an important feature for brewing of qualitatively good beer. Flocculation involves two main cell wall structures, which are the flocculation proteins (flocculins) and mannans, to which these flocculins bind. Unfortunately, in practice, the flocculation ability may get lost after several repitches. Flow cytometry was employed to analyze glucose and mannose structures of the cell surface by application of fluorescent lectins. Validation of the expression of the flocculin genes Lg-FLOl, FLO1, FLO5, and FLO9 was carried out using microarray techniques. SDS-PAGE, western blot, and ESI-MS/MS analyses served to isolate and determine yeast cell flocculins. Mannose and glucose labeling with fluorescent lectins allowed differentiating powdery and flocculent yeast cells under laboratory conditions. Using microarray techniques and proteomics, the four flocculation genes Lg-FLOl, FLO1, FLO5, FLO9, and the protein Lg-Flolp were identified as factors of major importance for flocculation. The expression of the genes was several times higher in flocculent yeast cells than in powdery ones. Flow cytometry is a fast and simple method to quantify the proportions of powdery and flocculent yeast cells in suspensions under defined cultivation conditions. However, differentiation under industrial conditions will require mRNA and protein expression profiling.
AB - The ability of brewing yeast to flocculate is an important feature for brewing of qualitatively good beer. Flocculation involves two main cell wall structures, which are the flocculation proteins (flocculins) and mannans, to which these flocculins bind. Unfortunately, in practice, the flocculation ability may get lost after several repitches. Flow cytometry was employed to analyze glucose and mannose structures of the cell surface by application of fluorescent lectins. Validation of the expression of the flocculin genes Lg-FLOl, FLO1, FLO5, and FLO9 was carried out using microarray techniques. SDS-PAGE, western blot, and ESI-MS/MS analyses served to isolate and determine yeast cell flocculins. Mannose and glucose labeling with fluorescent lectins allowed differentiating powdery and flocculent yeast cells under laboratory conditions. Using microarray techniques and proteomics, the four flocculation genes Lg-FLOl, FLO1, FLO5, FLO9, and the protein Lg-Flolp were identified as factors of major importance for flocculation. The expression of the genes was several times higher in flocculent yeast cells than in powdery ones. Flow cytometry is a fast and simple method to quantify the proportions of powdery and flocculent yeast cells in suspensions under defined cultivation conditions. However, differentiation under industrial conditions will require mRNA and protein expression profiling.
KW - Flocculation
KW - Lg-flolp
KW - Microarray
KW - Microbial flow cytometry
KW - Proteome
KW - Saccharomyces cerevisiae
UR - http://www.scopus.com/inward/record.url?scp=60849100218&partnerID=8YFLogxK
U2 - 10.1002/cyto.a.20661
DO - 10.1002/cyto.a.20661
M3 - Article
C2 - 19072835
AN - SCOPUS:60849100218
VL - 75
SP - 140
EP - 147
JO - Cytometry Part A
JF - Cytometry Part A
SN - 1552-4922
IS - 2
ER -