Kobayashi O, Hayashi N, Kuroki R, Sone H
Central Laboratories for Key Technology, Kirin Brewery Co., Ltd., 1-13-5 Fukuura, Kanazawa-ku, Yokohama-shi, Kanagawa 236-0004, Japan.
J Bacteriol. 1998 Dec;180(24):6503-10. doi: 10.1128/JB.180.24.6503-6510.1998.
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.
酵母絮凝是一种据信由类凝集素蛋白与位于酵母细胞表面的甘露糖链之间的相互作用导致的现象。编码细胞壁蛋白的FLO1基因被认为在酵母絮凝中起重要作用,这种絮凝被甘露糖抑制但不被葡萄糖抑制(甘露糖特异性絮凝)。从一种絮凝性底部发酵酵母菌株中分离出了一种新的FLO1同源物,命名为Lg - FLO1,在该菌株中絮凝被甘露糖和葡萄糖都抑制(甘露糖/葡萄糖特异性絮凝)。为了证实FLO1和Lg - FLO1都参与酵母絮凝现象,将甘露糖特异性絮凝菌株中的FLO1基因替换为Lg - FLO1基因。整合了Lg - FLO1基因的转化体表现出与甘露糖/葡萄糖特异性絮凝菌株相同的絮凝表型,这表明FLO1和Lg - FLO1基因分别编码甘露糖特异性和甘露糖/葡萄糖特异性的类凝集素蛋白。此外,通过表达嵌合FLO1和Lg - FLO1基因鉴定了这些糖的糖识别位点。发现这两种基因产物从氨基酸196到氨基酸240的区域对絮凝表型很重要。对该区域的进一步突变分析表明,Lg - Flo1蛋白中的苏氨酸 - 202和Flo1蛋白中的色氨酸 - 228参与糖识别。