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Appl Microbiol Biotechnol (2011) 90:377–384 DOI 10.1007/s00253-011-3115-8 BIOENERGY AND BIOFUELS Direct ethanol production from cassava pulp using a surface-engineered yeast strain co-displaying two amylases, two cellulases, and β-glucosidase Waraporn Apiwatanapiwat Yoshinori Murata Akihiko Kosugi Ryosuke Yamada Akihiko Kondo Takamitsu Arai Prapassorn Rugthaworn Yutaka Mori Received: 30 October 2010 /Revised: 9 January 2011 /Accepted: 9 January 2011 /Published online: 16 February 2011 # Springer-Verlag 2011 Abstract In order to develop a method for producing fuel fiber substrates contained in the cassava pulp decreased. ethanol from cassava pulp using cell surface engineering The results clearly showed that the arming yeast was able to (arming) technology, an arming yeast co-displaying α- produce ethanol directly from cassava pulp without amylase (α-AM), glucoamylase, endoglucanase, cellobio- addition of any hydrolytic enzymes. hydrase, and β-glucosidase on the surface of the yeast cells was constructed. The novel yeast strain, possessing the Keywords Arming yeast . Cell surface display . Ethanol . activities of all enzymes, was able to produce ethanol Amylases . Cassava pulp . Cellulases directly from soluble starch, barley β-glucan, and acid- treated Avicel. Cassava is a major crop in Southeast Asia and used mainly for starch production. In the starch Introduction manufacturing process, large amounts of solid wastes, called cassava pulp, are produced. The major components Fossil fuels have negative environmental effects because of cassava pulp are starch (approximately 60%) and the utilization of petroleum facilitates release of green cellulose fiber (approximately 30%). We attempted simul- h