Gene conferring flocculating property on yeast and gene product

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...

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435 41, 435 691, 4351723, 43525421, 4353201, 536 231, 536 237, 530300, 530324, 530327, 530350, C12N 1531, C12N 1581, C07K 700, C07K 14395

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058663746

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BRIEF SUMMARY
This is the U.S. national stage, filed under 35 U.S.C. 371, of PCT/JP96/00183 filed Jan. 31, 1996.
1. Field of the Invention
The present invention relates to a yeast flocculation gene and the use thereof. More specifically, the present invention relates to a protein having an activity of conferring on yeast brewer's yeast-type flocculating property, a DNA coding for the protein, a plasmid containing the DNA, a method for producing a yeast strain wherein brewer's yeast-type flocculating property has been conferred or enhanced with the use of the DNA, a method for producing a yeast strain wherein brewer's yeast-type flocculating property has been eliminated or reduced with the use of the DNA, and a method for eliminating or reducing brewer's yeast-type flocculating property of yeast by inhibiting the expression of the DNA.
The present invention also relates to a yeast strain wherein brewer's yeast-type flocculating property has been conferred, enhanced, eliminated or reduced by one of the above-mentioned methods.
Further, the present invention relates to a method for producing brewed products comprising culturing the above yeast strain as well as brewed products obtained by the method.
2. Background Art
It is a well known fact that, in alcoholic drinks such as beer and wine, the flocculating property of yeast used in fermentation is important not only because it determines the flavor and taste of the resultant products but also because it influences the workability in the fermentation process. The yeast which is used in the brewing of lager beer widely produced in Germany, Japan and many other countries has a characteristic that cells flocculate and sediment to the bottom of the fermented wort near the end of fermentation; such yeast is particularly called bottom fermenting yeast. In beer brewing, there is a characteristic feature in the process not found in other brewing, that is, yeast cells sedimented at the end of fermentation are recovered and reused in the subsequent fermentation. Accordingly, the characteristic of bottom fermenting yeast that it sediments at the late stage of fermentation has an especially big meaning for beer brewing.
Since brewer's yeast is one of the major factors that determine the quality of the final beer product, the breeding of excellent yeast strains is an important subject for beer brewers. In the breeding of bottom fermenting yeast, the conferring of a suitable flocculating property on yeast has a significant meaning. This is because a yeast strain too strong in flocculating property precipitates in the fermented wort during the fermentation, which leads to the termination of the fermentation, and, on the other hand, a yeast strain lacking flocculating property remains suspending even at the late stage of the fermentation and an operation such as centrifugation is needed to remove yeast cells from fermented wort. Therefore, a desirable yeast strain for the current production method is a strain which is dispersed at the beginning of the fermentation and precipitates well at the late stage of the fermentation. If the production method is different, needless to say, a yeast strain having a flocculating property suitable for the method is required.
In spite of the numerous studies concerning the yeast flocculating property which is industrially significant, the mechanism of yeast flocculation has not been elucidated yet. It is hard to say that control of flocculating property by improving a yeast strain per se is successful. As a result of years of genetic researches, the existence of yeast genes such as FLO1, flo3, FLO5, FLO8, sfl1, fsu1, fsu2, tup1, cyc8, cka2, FMC1 as well as the genes oli1 and oxi2 in mitochondrial DNA have been confirmed as genes involved in the flocculating property of yeast. As a study of these genes involved in the flocculating property of yeast at the molecular level, the (1993) and Yeast, 10, 211 (1994)!. Also, the isolation and analysis of the FLO5 gene has been reported. This report has shown that, although the location of the FLO5 gene is different

REFERENCES:
Teunissen et al., Sequence of the Open Reading Frame of the FLO1 Gene From Saccharomyces Cerevisiae, Yeast, (1993), V. 9, pp. 423-427.
Watari et al., Construction of Flocculent Brewers's Yeast by Chromosomal Integration of the Yeast Flocculation Gene FLO1, J. Inst. Brew., (1994), V. 100, pp. 73-77.
Teunissen et al., The Dominant Flocculation Genes of Saccharomyces Cerervisiae Constitute a New Subtelomeric Gene Family, Yeast, (1995), V. 11, pp. 1001-1013.
Murakami, Analysis of the Nucleotide Sequence of Chromosome VI From Saccharomyces Cerevisiae, Nature Genetics, (1995), V. 10, pp. 261-268.
Yamashita, Mating Signals Control Expression of Both Starch Fermentation Genes and a Novel Flocculation Gene FLO8 in the Yeast Saccharomyces, Agric. Biol. Chem., (1983), V. 47 (12), pp. 2889-2896.
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Watari, Breeding of Flocculent Industrial Saccharomyces Cerevisiae Strains by Introducing the Flocculation Gene FLO1, Agric. Biol. Chem., (1991), vol. 55 (6), pp. 1547-1552.
Watari, Molecular Cloning and Analysis of the Yeast Flocculation Gene FLO1, Yeast, (1994), V. 10, pp. 211-225.
Bidard, Cloning and Analysis of a FLO5 Flocculation Gene From S. Cerevisiae, Curr Genet, (1994), V. 25, pp. 196-201.
Sieiro, Flocculation of Industrial and Laboratory Strains of Saccharomyces Cerevisiae, J. Industrial Microbiology, (1995), V. 14, pp. 461-466.
Teunissen, Transcriptional Regulation of Flocculation Genes in Saccharomyces Cerevisiae, Yeast, (1995), V. 11, pp. 435-446.
By Brewing Society of Japan "Journal by Brewing Society of Japan", Sep. 15, 1993, Brewing Society of Japan, pp. 665-670.
Stewart, Can a Genetically Manipulated Yeast Strain Produce Palatable Beer?, J. Am. Soc. Brew. Chem., (1977), V. 35 (4), pp. 168-178.
Barney, Use of Genetic Transformation for the Introduction of Flocculence Into Yeast, J. Am. Soc. Brew. Chem., (1980), V. 38(2) pp. 71-74.

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