文献类型：期刊文献

英文题名：Yeast Surface Display of Trifunctional Minicellulosomes for Simultaneous Saccharification and Fermentation of Cellulose to Ethanol

作者：Wen, Fei[1];Sun, Jie[1,2];Zhao, Huimin[1,3,4,5]

机构：[1]Univ Illinois, Dept Chem & Biomol Engn, Urbana, IL 61801 USA;[2]E China Univ Sci & Technol, Dept Biol Engn, Shanghai 200237, Peoples R China;[3]Univ Illinois, Ctr Biophys & Computat Biol, Inst Genom Biol, Dept Chem, Urbana, IL 61801 USA;[4]Univ Illinois, Ctr Biophys & Computat Biol, Inst Genom Biol, Dept Biochem, Urbana, IL 61801 USA;[5]Univ Illinois, Ctr Biophys & Computat Biol, Inst Genom Biol, Dept Bioengn, Urbana, IL 61801 USA

年份：2010

卷号：76

期号：4

起止页码：1251

外文期刊名：APPLIED AND ENVIRONMENTAL MICROBIOLOGY

收录：;EI(收录号：20100712718974);WOS:【SCI-EXPANDED(收录号:WOS:000274328900033)】；

基金：We thank Nikhil U. Nair for helpful discussions and members of the Zhao laboratory for critical comments on the manuscript. We also thank Barbara Pilas and Bernard Montez at the Biotechnology Center and Alexander Ulanov at the Metabolomics Center of the University of Illinois for technical assistance.This work was supported by the Centennial Endowed Chair Fund of the Department of Chemical and Biomolecular Engineering at the University of Illinois at Urbana-Champaign.

语种：英文

外文关键词：Ethanol - Fermentation - Yeast - Cytology - Cell membranes - Cellulose - Saccharification

摘要：By combining cellulase production, cellulose hydrolysis, and sugar fermentation into a single step, consolidated bioprocessing (CBP) represents a promising technology for biofuel production. Here we report engineering of Saccharomyces cerevisiae strains displaying a series of uni-, bi-, and trifunctional minicellulosomes. These minicellulosomes consist of (i) a miniscaffoldin containing a cellulose-binding domain and three cohesin modules, which was tethered to the cell surface through the yeast a-agglutinin adhesion receptor, and (ii) up to three types of cellulases, an endoglucanase, a cellobiohydrolase, and a beta-glucosidase, each bearing a C-terminal dockerin. Cell surface assembly of the minicellulosomes was dependent on expression of the miniscaffoldin, indicating that formation of the complex was dictated by the high-affinity interactions between cohesins and dockerins. Compared to the unifunctional and bifunctional minicellulosomes, the quaternary trifunctional complexes showed enhanced enzyme-enzyme synergy and enzyme proximity synergy. More importantly, surface display of the trifunctional minicellulosomes gave yeast cells the ability to simultaneously break down and ferment phosphoric acid-swollen cellulose to ethanol with a titer of similar to 1.8 g/liter. To our knowledge, this is the first report of a recombinant yeast strain capable of producing cell-associated trifunctional minicellulosomes. The strain reported here represents a useful engineering platform for developing CBP-enabling microorganisms and elucidating principles of cellulosome construction and mode of action.