caa a22 4500 605503699 CHVBK 20210128100612.0 cr unu---uuuuu 210128e20150201xx s 000 0 eng 10.1007/s00253-014-6250-1 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6250-1 Efficient co-displaying and artificial ratio control of α-amylase and glucoamylase on the yeast cell surface by using combinations of different anchoring domains [Elektronische Daten] [Kentaro Inokuma, Takanobu Yoshida, Jun Ishii, Tomohisa Hasunuma, Akihiko Kondo] Recombinant yeast strains that display heterologous amylolytic enzymes on their cell surface via the glycosylphosphatidylinositol (GPI)-anchoring system are considered as promising biocatalysts for direct ethanol production from starchy materials. For the effective hydrolysis of these materials, the ratio optimization of multienzyme activity displayed on the cell surface is important. In this study, we have presented a ratio control system of multienzymes displayed on the yeast cell surface by using different GPI-anchoring domains. The novel gene cassettes for the cell-surface display of Streptococcus bovis α-amylase and Rhizopus oryzae glucoamylase were constructed using the Saccharomyces cerevisiae SED1 promoter and two different GPI-anchoring regions derived from Saccharomyces cerevisiae SED1 or SAG1. These gene cassettes were integrated into the Saccharomyces cerevisiae genome in different combinations. Then, the cell-surface α-amylase and glucoamylase activities and ethanol productivity of these recombinant strains were evaluated. The combinations of the gene cassettes of these enzymes affected the ratio of cell-surface α-amylase and glucoamylase activities and ethanol productivity of the recombinant strains. The highest ethanol productivity from raw starch was achieved by the strain harboring one α-amylase gene cassette carrying the SED1-anchoring region and two glucoamylase gene cassettes carrying the SED1-anchoring region (BY-AASS/GASS/GASS). This strain yielded 22.5 ± 0.6g/L of ethanol from 100g/L of raw starch in 120h of fermentation. Springer-Verlag Berlin Heidelberg, 2014 Saccharomyces cerevisiae nationallicence Cell-surface display nationallicence α-Amylase nationallicence Glucoamylase nationallicence Ratio control nationallicence Inokuma Kentaro Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, 657-8501, Kobe, Japan aut Yoshida Takanobu Technology Research Association of Highly Efficient Gene Design, Integrated Research Center of Kobe University 108, 7-1-48 Minatojimaminamimachi, Chuo-ku, 650-0047, Kobe, Japan aut Ishii Jun Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, 657-8501, Kobe, Japan aut Hasunuma Tomohisa Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, 657-8501, Kobe, Japan aut Kondo Akihiko Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, 657-8501, Kobe, Japan aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/4(2015-02-01), 1655-1663 0175-7598 99:4<1655 2015 99 253 https://doi.org/10.1007/s00253-014-6250-1 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6250-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Inokuma Kentaro Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, 657-8501, Kobe, Japan aut NATIONALLICENCE 700 1- Yoshida Takanobu Technology Research Association of Highly Efficient Gene Design, Integrated Research Center of Kobe University 108, 7-1-48 Minatojimaminamimachi, Chuo-ku, 650-0047, Kobe, Japan aut NATIONALLICENCE 700 1- Ishii Jun Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, 657-8501, Kobe, Japan aut NATIONALLICENCE 700 1- Hasunuma Tomohisa Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodai, Nada, 657-8501, Kobe, Japan aut NATIONALLICENCE 700 1- Kondo Akihiko Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, 657-8501, Kobe, Japan aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/4(2015-02-01), 1655-1663 0175-7598 99:4<1655 2015 99 253