Biotechnological products and process engineering

Applied Microbiology and Biotechnology

, Volume 98, Issue 3, pp 1087-1094

Analysis of cellodextrin transporters from Neurospora crassa in Saccharomyces cerevisiae for cellobiose fermentation

  • Heejin KimAffiliated withDepartment of Food Science and Human Nutrition, University of Illinois at Urbana-ChampaignInstitute for Genomic Biology, University of Illinois at Urbana-Champaign
  • , Won-Heong LeeAffiliated withDepartment of Food Science and Human Nutrition, University of Illinois at Urbana-ChampaignInstitute for Genomic Biology, University of Illinois at Urbana-Champaign
  • , Jonathan M. GalazkaAffiliated withDepartment of Molecular and Cell Biology, University of California at BerkeleyPhysical Biosciences Division, Lawrence Berkeley National Laboratory
  • , Jamie H. D. CateAffiliated withDepartment of Molecular and Cell Biology, University of California at BerkeleyPhysical Biosciences Division, Lawrence Berkeley National Laboratory
  • , Yong-Su JinAffiliated withDepartment of Food Science and Human Nutrition, University of Illinois at Urbana-ChampaignInstitute for Genomic Biology, University of Illinois at Urbana-Champaign Email author 

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Abstract

Saccharomyces cerevisiae can be engineered to ferment cellodextrins produced by cellulases as a product of cellulose hydrolysis. Direct fermentation of cellodextrins instead of glucose is advantageous because glucose inhibits cellulase activity and represses the fermentation of non-glucose sugars present in cellulosic hydrolyzates. To facilitate cellodextrin utilization by S. cerevisiae, a fungal cellodextrin-utilizing pathway from Neurospora crassa consisting of a cellodextrin transporter and a cellodextrin hydrolase has been introduced into S. cerevisiae. Two cellodextrin transporters (CDT-1 and CDT-2) were previously identified in N. crassa, but their kinetic properties and efficiency for cellobiose fermentation have not been studied in detail. In this study, CDT-1 and CDT-2, which are hypothesized to transport cellodextrin with distinct mechanisms, were introduced into S. cerevisiae along with an intracellular β-glucosidase (GH1-1). Cellobiose transport assays with the resulting strains indicated that CDT-1 is a proton symporter while CDT-2 is a simple facilitator. A strain expressing CDT-1 and GH1-1 (DCDT-1G) showed faster cellobiose fermentation than the strain expressing CDT-2 and GH1-1 (DCDT-2G) under various culture conditions with different medium compositions and aeration levels. While CDT-2 is expected to have energetic benefits, the expression levels and kinetic properties of CDT-1 in S. cerevisiae appears to be optimum for cellobiose fermentation. These results suggest CDT-1 is a more effective cellobiose transporter than CDT-2 for engineering S. cerevisiae to ferment cellobiose.

Keywords

Cellulosic ethanol Cellodextrin transporters Intracellular β-glucosidase Engineered S. cerevisiae