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Journal of Bacteriology, December 1998, p. 6503-6510, Vol. 180, No. 24
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Region of Flo1 Proteins Responsible for Sugar Recognition

Osamu Kobayashi,1,* Nobuyuki Hayashi,2 Ryota Kuroki,1 and Hidetaka Sone1

Central Laboratories for Key Technology, Kirin Brewery Co., Ltd., 1-13-5 Fukuura, Kanazawa-ku, Yokohama-shi, Kanagawa 236-0004,1 and Research Laboratory for Brewing, Kirin Brewery Co., Ltd., 1-17-1, Namamugi, Tsurumi-ku, Yokohama-shi, Kanagawa 230-8628,2 Japan

Received 29 May 1998/Accepted 16 October 1998

Yeast flocculation is a phenomenon which is believed to result from an interaction between a lectin-like protein and a mannose chain located on the yeast cell surface. The FLO1 gene, which encodes a cell wall protein, is considered to play an important role in yeast flocculation, which is inhibited by mannose but not by glucose (mannose-specific flocculation). A new homologue of FLO1, named Lg-FLO1, was isolated from a flocculent bottom-fermenting yeast strain in which flocculation is inhibited by both mannose and glucose (mannose/glucose-specific flocculation). In order to confirm that both FLO1 and Lg-FLO1 are involved in the yeast flocculation phenomenon, the FLO1 gene in the mannose-specific flocculation strain was replaced by the Lg-FLO1 gene. The transformant in which the Lg-FLO1 gene was incorporated showed the same flocculation phenotype as the mannose/glucose-specific flocculation strain, suggesting that the FLO1 and Lg-FLO1 genes encode mannose-specific and mannose/glucose-specific lectin-like proteins, respectively. Moreover, the sugar recognition sites for these sugars were identified by expressing chimeric FLO1 and Lg-FLO1 genes. It was found that the region from amino acid 196 to amino acid 240 of both gene products is important for flocculation phenotypes. Further mutational analysis of this region suggested that Thr-202 in the Lg-Flo1 protein and Trp-228 in the Flo1 protein are involved in sugar recognition.

* Corresponding author. Mailing address: Central Laboratories for Key Technology, Kirin Brewery Co., Ltd., 1-13-5 Fukuura, Kanazawa-ku, Yokohama-shi, Kanagawa 236-0004, Japan. Phone: 81-45-788-7215. Fax: 81-45-788-4042. E-mail: osamuk{at}kirin.co.jp.

Journal of Bacteriology, December 1998, p. 6503-6510, Vol. 180, No. 24
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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