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Improving genetic immobilization of a cellulase on yeast cell surface for bioethanol production using cellulose
Author(s) -
Yang Jinying,
Dang Hongyue,
Lu Jian Ren
Publication year - 2013
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.201100602
Subject(s) - cellulase , saccharomyces cerevisiae , yeast , biochemistry , cellulose , clostridium thermocellum , gene , bacillus subtilis , biology , fermentation , chemistry , genetics , bacteria
In this study, Saccharomyces cerevisiae was genetically engineered to harbor the capability of utilizing celluloses for bioethanol production by displaying active cellulolytic enzymes on the cell surface. An endo‐1,4‐β‐glucanase gene egX was cloned from Bacillus pumilus C‐9 and its expression products, the EGX cellulases, were displayed on the cell surface of S. cerevisiae by fusing egX with aga2 that encodes the binding subunit of the S. cerevisiae cell wall protein α‐agglutinin. To achieve high gene copies and stability, multicopy integration was obtained by integrating the fusion aga2 ‐ egX gene into the rDNA region of the S. cerevisiae chromosome. To achieve high expression and surface display efficiency, the aga2 ‐ egX gene was expressed under the control of a strong promoter. The presence of the enzymatically active cellulase fusion proteins on the S. cerevisiae cell surface was verified by carboxymethyl cellulase activity assay and immunofluorescence microscopy. This work presented a promising strategy to genetically engineer yeasts to perform efficient fermentation of cellulosic materials for bioethanol production.