Immobilization Of Enzyme On Yeast Cell Surface

To immobilize protein on the cell surface of S. cerevisiae, molecular information of the native cell wall-anchored protein, «-agglutinin, was utilized. «-Agglutinin (1) is a mannoprotein involved in the sexual adhesion of mating-type « S. cerevisiae cells with mating-type a S. cerevisiae cells. «-Agglutinin has a glycosylphospha-tidylinositol (GPI) anchor attachment signal (Fig. 1; Table 1), which is involved in anchoring cell wall proteins. This anchoring signal was combined with the

Figure 1 Transportation system of cell surface proteins in S. cerevisiae and GPI anchor structure. ER, endoplasmic reticulum; PI-PLC, phosphatidylinositol-specific phospholipase C; GPI, glycosylphosphatidylinositol; Man, mannose; GlcNH2, glucosamine.

signal of the secreted enzymes using genetic engineering techniques. Figure 2 shows the general structure of the gene for cell surface immobilization of an enzyme (2). 3'-Half of «-agglutinin contains a glycosylphosphatidyl-inositol (GPI) anchor attachment signal at the C-term-inal end, like other cell surface proteins.

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