Enhanced cell-surface display of a heterologous protein using &ITSED1&IT anchoring system in &ITSED1&IT-disrupted &ITSaccharomyces cerevisiae&IT strain

被引:19
作者
Bamba, Takahiro [1 ]
Inokuma, Kentaro [2 ]
Hasunuma, Tomohisa [2 ]
Kondo, Akihiko [2 ,3 ]
机构
[1] Kobe Univ, Grad Sch Engn, Dept Chem Sci & Engn, 1-1 Rokkodai, Kobe, Hyogo 6578501, Japan
[2] Kobe Univ, Grad Sch Sci Technol & Innovat, 1-1 Rokkodai, Kobe, Hyogo 6578501, Japan
[3] RIKEN, Biomass Engn Program, Tsurumi Ku, 1-7-22 Suehiro Cho, Yokohama, Kanagawa 2300045, Japan
关键词
Saccharomyces cerevisiae; Cell surface display; beta-Glucosidase; SED1; Mass spectrometry; Aspergillus aculeatus; SACCHAROMYCES-CEREVISIAE; ETHANOL-PRODUCTION; STATIONARY-PHASE; ALPHA-AMYLASE; YEAST; GLUCOAMYLASE; WALL; BIOSYNTHESIS; FERMENTATION; EXPRESSION;
D O I
10.1016/j.jbiosc.2017.09.013
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Yeast displaying enzymes on the cell surface are used for developing whole-cell biocatalysts. High enzyme activity on the cell surface is required in certain applications such as direct ethanol production from lignocellulosic materials. However, the cell surface enzyme activity is limited by several factors, one of which is the protein amount of the yeast cell wall. In this study, we attempted to improve the incorporation capacity of a displayed heterologous enzyme by disrupting a native cell-wall protein. beta-Glucosidase (BGL1) from Aspergillus aculeatus was fused with Saccharomyces cerevisiae Sedi and displayed on the cell surface of S. cerevisiae BY4741 strain and its SED1 disruptant. Sedi is one of the most abundant stationary phase yeast cell wall protein. A time course analysis revealed that BGLI activity of the control strain reached saturation after 48 h of cultivation. In contrast, the BGL1 activity of the SED1 disruptant increased until 72 h of cultivation and was 22% higher than that of the control strain. We also performed relative quantification of cell wall proteins of these strains by nanoscale ultra pressure liquid chromatography electrospray ionization quadrupole time-of-flight tandem mass spectrometry (nano-UPLC-MSE). The amount of the cell wall-associated BGL1 per unit dry cell-weight of the SED1 disruptant was 19% higher than that of the control strain. These results suggested that the incorporation capacity of the cell wall for BGL1 was increased by disruption of SED1. Disruption of SED1 would be a promising approach for improving display efficiency of heterologous protein fused with Sed1. (C) 2017, The Society for Biotechnology, Japan. All rights reserved.
引用
收藏
页码:306 / 310
页数:5
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