title: Cell wall structure suitable for surface display of proteins in Saccharomyces cerevisiae.
abstract: A display system for adding new protein functions to the cell surfaces of microorganisms has been developed, and applications of the system to various fields have been proposed. With the aim of constructing a cell surface environment suitable for protein display in Saccharomyces cerevisiae, the cell surface structures of cell wall mutants were investigated. Four cell wall mutant strains were selected by analyses using a GFP display system via a GPI anchor. β-Glucosidase and endoglucanase II were displayed on the cell surface in the four mutants, and their activities were evaluated. mnn2 deletion strain exhibited the highest activity for both the enzymes. In particular, endoglucanase II activity using carboxymethylcellulose as a substrate in the mutant strain was 1.9-fold higher than that of the wild-type strain. In addition, the activity of endoglucanase II released from the mnn2 deletion strain by Zymolyase 20T treatment was higher than that from the wild-type strain. The results of green fluorescent protein (GFP) and endoglucanase displays suggest that the amounts of enzyme displayed on the cell surface were increased by the mnn2 deletion. The enzyme activity of the mnn2 deletion strain was compared with that of the wild-type strain. The relative value (mnn2 deletion mutant/wild-type strain) of endoglucanase II activity using carboxymethylcellulose as a substrate was higher than that of β-glucosidase activity using p-nitrophenyl-β-glucopyranoside as a substrate, suggesting that the cell surface environment of the mnn2 deletion strain facilitates the binding of high-molecular-weight substrates to the active sites of the displayed enzymes.
authors.affiliation: Department of Applied Chemistry and Bioengineering, Graduate School of Engineering, Osaka City University, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Matsuoka
authors.forename: Kazuya
authors.initials: K
authors.lastname: Hashimoto
authors.forename: Aki
authors.initials: A
authors.lastname: Saijo
authors.forename: Yuki
authors.initials: Y
authors.lastname: Takada
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kondo
authors.forename: Mitsuyoshi
authors.initials: M
authors.lastname: Ueda
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ooshima
authors.forename: Taro
authors.initials: T
authors.lastname: Tachibana
authors.forename: Masayuki
authors.initials: M
authors.lastname: Azuma
mesh_terms: D002473:Cell Wall / Q000737:chemistry* / Q000201:enzymology* / Q000235:genetics; D002480:Cellulase / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D008364:Mannosyltransferases / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000201:enzymology / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D001617:beta-Glucosidase / Q000235:genetics / Q000378:metabolism*
chemical_list: D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D049452:Green Fluorescent Proteins; C090871:MNN2 protein, S cerevisiae; D008364:Mannosyltransferases; C056373:endoglucanase 2; D001617:beta-Glucosidase; D002480:Cellulase
keywords: MNN2; cell surface; cell wall; glucan; mannan