title: Display of active beta-glucosidase on the surface of Schizosaccharomyces pombe cells using novel anchor proteins.
abstract: Here, we demonstrate display of beta-glucosidase (BGL) on the surface of Schizosaccharomyces pombe cells using novel anchor proteins. A total of four candidate anchor proteins (SPBC21D10.06c, SPBC947.04, SPBC19C7.05, and SPBC359.04c) were selected from among almost all of S. pombe membrane proteins. The C-terminus of each anchor protein was genetically fused to the N-terminus of BGL, and the fusion protein was expressed using S. pombe as a host. The highest cell surface-associated BGL activity (107 U/10(5) cells was achieved with SPBC359.04c serving as the anchor, followed by SPBC947.04 (44 U/10(5) cells) and SPBC21D10.06c (38 U/10(5) cells). S. pombe displaying BGL with SPBC359.04c as an anchor showed the highest growth on 2 % cellobiose (10.7 × 10(7) cells/mL after 41 h of cultivation from an initial density of 0.1 × 10(7) cells/mL). Additionally, culturing BGL-displaying S. pombe in medium containing cellobiose as the sole carbon source did not affect protein expression, and ethanol fermentation from cellobiose was successfully demonstrated using BGL-displaying S. pombe. This is the first report describing a cell surface display system for the functionalization of S. pombe.
authors: Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1, Rokkodai-cho, Nada, Kobe 657-8501, Japan.
authors: Tsutomu
authors: T
authors: Tanaka
authors: Sayoko
authors: S
authors: Matsumoto
authors: Mari
authors: M
authors: Yamada
authors: Ryosuke
authors: R
authors: Yamada
authors: Fumio
authors: F
authors: Matsuda
authors: Akihiko
authors: A
authors: Kondo
mesh_terms: D001483:Base Sequence; D002475:Cellobiose / Q000378:metabolism; D017931:DNA Primers; D005285:Fermentation; D008969:Molecular Sequence Data; D010957:Plasmids; D012568:Schizosaccharomyces / Q000201:enzymology*; D001617:beta-Glucosidase / Q000378:metabolism*
chemical_list: D017931:DNA Primers; D002475:Cellobiose; D001617:beta-Glucosidase