Project 2A - A platform for lignocellulose active enzyme and accessory protein production
PI - Reg Stroms
Objectives: This project focuses on the production of lignocellulose-active enzymes and accessory proteins for biorefinery applications. The objectives are to improve the expression level and purity of lignocellulose active enzyme and accessory protein production by Aspergillus niger, a filamentous fungus widely used for secreted protein production. To achieve this objective we will pursue the following three complementary approaches: 1) use transcription-profiling data to develop rational strategies to improve the protein secretion capabilities of A. niger, 2) develop expression vectors that improve target protein production through the use of new promoters and secretion signals, and 3) create clean expression strains that do not produce "contaminating secreted proteins" that interfere with the production or activity of the expressed heterologous lignocellulose-active proteins. The successful combination of these three approaches will enable A. niger to express essentially pure lignocellulose active enzymes and accessory proteins at levels compatible with industrial processing without the need for costly and time consuming post expression protein purification.
José Humberto Tambor, Huanan Ren, Sophia Ushinsky, Yun Zheng, Anja Riemens, Christopher St-Francois, Adrian Tsang, Justin Powlowski and Reginald Storms. Recombinant expression, activity screening and functional characterization identifies three novel endo-1,4-beta-glucanases that efficiently hydrolyse cellulosic substrates Applied Microbiology and Biotechnology published 2012, 93:203-214.
Caroline Wilde, Nicholas Gold, Nancy Bawa, Jose Humberto Tambor, Lina Mougharbel, Reginald Storms and Vincent Martin. Expression of a library of beta-glucosidases in Saccharomyces cerevisiae for the development of a biomass fermenting strain. Applied Microbiology and Biotechnology accepted 2012 95:647-659.
Berka RM, Grigoriev IV, Otillar R,, Salamov A, Grimwood J, Reid I, Ishmael N, John T, Darmond C, Moisan M-C, Henrissat B, Coutinho PM, Lombard V, Natvig DO, Lindquist E2, Schmutz J, Lucas S, Harris P1, Powlowski J, Bellemare A, Taylor D, Butler G, de Vries RP, Allijn IE, van den Brink J, Ushinsky S, Storms R, Powell AJ, Paulsen IT, Elbourne LDH, Baker SE, Magnuson J, LaBoissiere S, Clutterbuck AJ, Martinez D, Wogulis M, de Leon AL, Michael W. Rey MW, and Tsang A. Comparative Genomic Analysis of the Thermophilic Biomass-Degrading Fungi Myceliophthora thermophila and Thielavia terrestris Nature Biotechnology 2011 Oct 2;29(10):922-927.