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Published on: October 24, 2016
Transporter engineering for cellobiose fermentation under lower pH conditions by engineered Saccharomyces cerevisiae
Eun Joong Oh1, Suryang Kwak1, Heejin Kim1
1Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Abstract:
The aim of this study was to engineer cellodextrin transporter 2 (CDT-2) from Neurospora crassa for improved cellobiose fermentation under lower pH conditions by Saccharomyces cerevisiae. Through directed evolution, a mutant CDT-2 capable of facilitating cellobiose fermentation under lower pH conditions was obtained. Specifically, a library of CDT-2 mutants with GFP fusion was screened by flow cytometry and then serial subcultured to isolate a CDT-2 mutant capable of transporting cellobiose under acidic conditions. The engineered S. cerevisiae expressing the isolated mutant CDT-2 (I96N/T487A) produced ethanol with a specific cellobiose consumption rate of 0.069g/gcell/h, which was 51% and 55% higher than those of the strains harboring wild-type CDT-1 and CDT-2 in a minimal medium with 2g/L of acetic acid.
