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Appl Microbiol Biotechnol

Title:		Genetic immobilization of cellulase on the cell surface of Saccharomyces cerevisiae
Author(s):		Murai T; Ueda M; Atomi H; Shibasaki Y; Kamasawa N; Osumi M; Kawaguchi T; Arai M; Tanaka A;
Address:		"Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Japan"
Journal Title:		Appl Microbiol Biotechnol
Year:		1997
Volume:		48
Issue:		4
Page Number:		499 - 503
DOI:		10.1007/s002530051086
ISSN/ISBN:		0175-7598 (Print) 0175-7598 (Linking)
Abstract:		"We tried genetically to immobilize cellulase protein on the cell surface of the yeast Saccharomyces cerevisiae in its active form. A cDNA encoding FI-carboxymethylcellulase (CMCase) of the fungus Aspergillus aculeatus, with its secretion signal peptide, was fused with the gene encoding the C-terminal half (320 amino acid residues from the C terminus) of yeast alpha-agglutinin a protein involved in mating and covalently anchored to the cell wall. The plasmid constructed containing this fusion gene was introduced into S. cerevisiae and expressed under the control of the glyceraldehyde-3-phosphate dehydrogenase promoter from S. cerevisiae. The CMCase activity was detected in the cell pellet fraction. The CMCase protein was solubilized from the cell wall fraction by glucanase treatment but not by sodium dodecyl sulphate treatment, indicating the covalent binding of the fusion protein to the cell wall. The appearance of the fused protein on the cell surface was further confirmed by immunofluorescence microscopy and immunoelectron microscopy. These results proved that the CMCase was anchored on the cell wall in its active form"
Keywords:		"Aspergillus/*enzymology Cell Wall/*enzymology Cellulase/genetics/*metabolism *Enzymes, Immobilized Fungal Proteins/genetics/*metabolism Genes, Synthetic Glyceraldehyde-3-Phosphate Dehydrogenases/genetics Mating Factor Microscopy, Fluorescence Microscopy, ;"
Notes:		"MedlineMurai, T Ueda, M Atomi, H Shibasaki, Y Kamasawa, N Osumi, M Kawaguchi, T Arai, M Tanaka, A eng Germany 1997/12/09 Appl Microbiol Biotechnol. 1997 Oct; 48(4):499-503. doi: 10.1007/s002530051086"