Yeast agglutination genes and yeast containing them

Chemistry: molecular biology and microbiology – Micro-organism – per se ; compositions thereof; proces of... – Fungi

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536 231, 536 232, 536 2374, 435 691, 435 711, 4351723, 43525411, 43525421, 4352551, 4352552, 435261, C12N 1531, C12N 1581, C12N 119

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055852718

DESCRIPTION:

BRIEF SUMMARY
TECHNICAL FIELD

The present invention relates to agglutination genes of agglutinative yeast, and to yeast which contains them.


BACKGROUND ART

In the fermentation industry, yeast agglutination is an industrially important phenomenon, and the use of agglutinative yeast is being studied while much research is being undertaken to discover the cause of its agglutination. Yeast agglutination is known to be controlled by a plurality of genes, a relatively well researched example thereof being the agglutination gene called FL01, which is mapped on the right arm of yeast chromosome I.
Regarding the structure of the agglutination gene FL01 derived from the yeast Saccharomyces cerevisiae, it had been completely unknown, but in 1989 it was cloned for the first time by the present inventors et al., and its restriction enzyme cleavage map has been determined (Watari et al., Agricultural and Biological Chemistry, Vol. 53, No. 3, p.901-903, 1989). (Nevertheless, it base sequence was unknown).
We the present inventors reported that it is possible to convert non-agglutinative industrial yeasts into agglutinative yeasts for practical use by the introduction of the agglutination gene FL01 into various industrial yeasts (Watari et al., Agricultural and Biological Chemistry, Vol. 55, No. 6, p.1547-1552, 1991); however, it was not always possible to impart strong and stable agglutinative properties to all of the industrial yeasts.
We the present inventors thereafter diligently pursued research on the FL01 gene, and discovered that the gene that we the present inventors et al. had reported as being the FL01 gene (Watari et al., Agricultural and Biological Chemistry, Vol. 53, No. 3, p. 901-903, 1989) was not the intact FL01 gene as present on chromosome I of the yeast Saccharomyces cerevisiae strain ABXL-1D, but was the FL01 gene with a portion thereof deleted during maintenance of the plasmid containing the intact FL01 gene in Escherichia coli strain K12 (hereunder, this gene shall be referred to as FL01S).


DISCLOSURE OF INVENTION

The object of the present invention is to establish the structure of the intact FL01 gene (hereunder, this gene shall be referred to as FL01L), and to provide a technique for imparting stronger and more stable agglutinative properties to various industrial yeasts.
Now, we the present inventors, as the result of varied research regarding the FL01 gene, have succeeded in isolating the intact FL01 gene, or FL01L gene, have determined the entire base sequence of the gene, and further have discovered that by using the FL01L gene, it is possible to breed various yeasts for practical use which have stronger and more stable agglutinative ability, compared with using the FL01S gene, and thus the present invention has been completed.
In other words, the present invention relates to an agglutination gene of 4.7.+-.0.2 kb in yeast which codes for a polypeptide which exhibits agglutinative activity, and specifically, it relates to the above mentioned agglutination gene which is derived from the yeast Saccharomyces cerevisiae and is defined by the restriction enzyme cleavage map in FIG. 1, and more specifically, it relates to the above mentioned agglutination gene which substantially codes for the amino acid sequence listed as Sequence No. 1 (SEQ ID No: 1).
The present invention also relates to an agglutination gene of 2.6.+-.0.2 kb in yeast which codes for a polypeptide which exhibits agglutinative activity, and specifically to an agglutination gene which is derived from the yeast Saccharomyces cerevisiae and is defined by the restriction enzyme cleavage map in FIG. 2, and more specifically, to an agglutination gene which substantially codes for the amino acid sequence listed as Sequence No. 3 (SEQ ID No: 3).
The present invention further relates to yeasts containing either of the above mentioned agglutination genes and having agglutinative properties.
"Agglutination gene" as mentioned in the present specification is used to mean a gene which controls agglutination of yeast.
As described above, the agglutinat

REFERENCES:
J. Warari et al. "Breeding of Flocculent Brewer's Yeast by Genetic Engineering", Proc. Congr. Eur. Brewery Convention, 23rd, 297-304 (1991).
A. W. Teunissen et al. "Northern Studies of the FL01 Gene of Saccharomyces cerevisiae", Yeast 8, Spec. Iss. S621 (1992).
Yeast, vol. 9, No. 4, Apr. 1993, A. W. R. H. Teunissen, et al., "Sequence of the Open Reading Frame of the FL01 Gene from Saccharomyces cerevisiae", pp. 423-427.
Agric. Biol. Chem., vol. 55, No. 6, 1991, pp. 1547-1552, Junji Watari, et al., "Breeding of Flocculent Industrial Saccharomyces cerevisiae Strains by Introducing the Flocculation Gene FL01".
Agricultural and Biological Chemistry, vol. 53, No. 3, Jan. 1989, Junji Watari, et al., "Molecular Cloning of a Flocculation Gene in Saccharomyces cerevisiae", pp. 901-903.
Yeast, vol. 9, No. 1, Jan. 1993, Aloys W. R. H. Teunissen, et al., "Physical Localization of the Flocculation Gene FL01 on Chromosome I of Saccharomyces cerevisiae", pp. 1-10.

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