RRC ID 48069
Author Inokuma K, Bamba T, Ishii J, Ito Y, Hasunuma T, Kondo A.
Title Enhanced cell-surface display and secretory production of cellulolytic enzymes with Saccharomyces cerevisiae Sed1 signal peptide.
Journal Biotechnol Bioeng
Abstract Recombinant yeast strains displaying aheterologous cellulolytic enzymes on their cell surfaces using a glycosylphosphatidylinositol (GPI) anchoring system are a promising strategy for bioethanol production from lignocellulosic materials. A crucial step for cell wall localization of the enzymes is the intracellular transport of proteins in yeast cells. Therefore, the addition of a highly efficient secretion signal sequence is important to increase the amount of the enzymes on the yeast cell surface. In this study, we demonstrated the effectiveness of a novel signal peptide (SP) sequence derived from the Saccharomyces cerevisiae SED1 gene for cell-surface display and secretory production of cellulolytic enzymes. Gene cassettes with SP sequences derived from S. cerevisiae SED1 (SED1SP), Rhizopus oryzae glucoamylase (GLUASP), and S. cerevisiae α-mating pheromone (MFα1SP) were constructed for cell-surface display of Aspergillus aculeatus β-glucosidase (BGL1) and Trichoderma reesei endoglucanase II (EGII). These gene cassettes were integrated into the S. cerevisiae genome. The recombinant strains with the SED1SP showed higher cell-surface BGL and EG activities than those with the conventional SP sequences (GLUASP and MFα1SP). The novel SP sequence also improved the secretory production of BGL and EG in S. cerevisiae. The extracellular BGL activity of the recombinant strains with the SED1SP was 1.3- and 1.9-fold higher than the GLUASP and MFα1SP strains, respectively. Moreover, the utilization of SED1SP successfully enhanced the secretory production of BGL in Pichia pastoris. The utilization of the novel SP sequence is a promising option for highly efficient cell-surface display and secretory production of heterologous proteins in various yeast species. Biotechnol. Bioeng. 2016;113: 2358-2366. © 2016 Wiley Periodicals, Inc.
Volume 113(11)
Pages 2358-66
Published 2016-11-1
DOI 10.1002/bit.26008
PMID 27183011
MeSH Cell Membrane / metabolism* Cellulase / metabolism* Genetic Enhancement / methods* Membrane Glycoproteins / genetics* Membrane Glycoproteins / metabolism Protein Engineering / methods Protein Transport / genetics Recombinant Proteins / biosynthesis* Recombinant Proteins / genetics Saccharomyces cerevisiae / physiology* Saccharomyces cerevisiae Proteins / genetics* Saccharomyces cerevisiae Proteins / metabolism
IF 4.002
Times Cited 18
Yeast pGK403