TY - JOUR
T1 - Use of PCR-DHPLC (polymerase chain reaction-denaturing high performance liquid chromatography) for the rapid differentiation of industrial Saccharomyces pastorianus and Saccharomyces cerevisiae strains
AU - Hutzler, M.
AU - Geiger, E.
AU - Jacob, F.
PY - 2010
Y1 - 2010
N2 - Strain specific detection and control of Saccharomyces pastorianus and Saccharomyces cerevisiae starter cultures is of great importance for the fermentation industry. The preconditions of strain specific fermentation characteristics can be ensured by periodic analysis and confirmation of the strain identity. With regard to industrial S. pastorianus and S. cerevisiae strains and a focus on brewing strains, the differentiation methods most available are time-consuming and not very discriminative. In this work PCR-DHPLC analysis was investigated as a novel approach for the differentiation of industrially used S. pastorianus and S. cerevisiae strains. The PCR-DHPLC-system was specific for S. cerevisiae strains and S. pastorianus hybrid strains that contain IGS2 rDNA, which originates from the S. cerevisiae ancestor. For the DNA of 177 strains of 41 non-target species, which are typical for beverage and fermentation surroundings, the absence of PCR-amplificates could be confirmed by DHPLC analysis. It was shown that single strains of S. cerevisiae and S. pastorianus could be differentiated. A strain specific differentiation within the group of top-fermenting Saccharomyces cerevisiae strains could also be performed. For the group of bottom fermenting S. pastorianus brewing strains, strain-to-strain specific differences in the DHPLC chromatograms could be observed which can be used to differentiate and to compare two single strains with each other, although the comparison of chromatograms of an unknown S. pastorianus strain with a set of known S. pastorianus chromatograms could only reveal tendencies towards grouping into types. The differential DHPLC chromatogram characteristics (fluorescence intensities, number of peaks/side-peaks/peak-shoulders) within S. pastorianus are present, but not as distinctive as for S. cerevisiae. Additionally PCR-DHPLC has advantages compared to other differentiation methods, such as species specificity, speed (2.5 h for one sample) and precision with the described limits.
AB - Strain specific detection and control of Saccharomyces pastorianus and Saccharomyces cerevisiae starter cultures is of great importance for the fermentation industry. The preconditions of strain specific fermentation characteristics can be ensured by periodic analysis and confirmation of the strain identity. With regard to industrial S. pastorianus and S. cerevisiae strains and a focus on brewing strains, the differentiation methods most available are time-consuming and not very discriminative. In this work PCR-DHPLC analysis was investigated as a novel approach for the differentiation of industrially used S. pastorianus and S. cerevisiae strains. The PCR-DHPLC-system was specific for S. cerevisiae strains and S. pastorianus hybrid strains that contain IGS2 rDNA, which originates from the S. cerevisiae ancestor. For the DNA of 177 strains of 41 non-target species, which are typical for beverage and fermentation surroundings, the absence of PCR-amplificates could be confirmed by DHPLC analysis. It was shown that single strains of S. cerevisiae and S. pastorianus could be differentiated. A strain specific differentiation within the group of top-fermenting Saccharomyces cerevisiae strains could also be performed. For the group of bottom fermenting S. pastorianus brewing strains, strain-to-strain specific differences in the DHPLC chromatograms could be observed which can be used to differentiate and to compare two single strains with each other, although the comparison of chromatograms of an unknown S. pastorianus strain with a set of known S. pastorianus chromatograms could only reveal tendencies towards grouping into types. The differential DHPLC chromatogram characteristics (fluorescence intensities, number of peaks/side-peaks/peak-shoulders) within S. pastorianus are present, but not as distinctive as for S. cerevisiae. Additionally PCR-DHPLC has advantages compared to other differentiation methods, such as species specificity, speed (2.5 h for one sample) and precision with the described limits.
UR - http://www.scopus.com/inward/record.url?scp=79952497837&partnerID=8YFLogxK
U2 - 10.1002/j.2050-0416.2010.tb00798.x
DO - 10.1002/j.2050-0416.2010.tb00798.x
M3 - Article
AN - SCOPUS:79952497837
SN - 0046-9750
VL - 116
SP - 464
EP - 474
JO - Journal of the Institute of Brewing
JF - Journal of the Institute of Brewing
IS - 4
ER -