Author

Ruoyu Yan

Date of Award

2011

Degree Type

Thesis

Degree Name

Master of Science

Program

Biology

Supervisor

Dr. Rima Menassa

Second Advisor

Dr. Susanne Kohalmi

Abstract

Lack of effective biomass pretreatments has been one of the main limiting factors for cellulosic ethanol production. The addition of plant-made hemicellulases to the pretreatment procedure is an alternative method of obtaining fermentable sugars. Four putative or characterized xylanases out of 73 hydrolases from A. niger were found by bioinformatics analysis. Proteins of interest were targeted to the ER, and transiently expressed in N. benthamiana plants accumulating up to 38% of TSP. A simple surfactant- based aqueous two-phase system was used to purify xylanase-HFBI fusions with a recovery up to 83%. An endoxylanase and a P-xylosidase were characterized, and their combined action enhanced xylose production. This study presents a very promising expression and purification system for fungal hemicellulases to further improve cellulosic ethanol production.

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