Directed evolution of a cellodextrin transporter for improved biofuel production under anaerobic conditions in Saccharomyces cerevisiae
Article first published online: 11 MAR 2014
© 2014 Wiley Periodicals, Inc.
Biotechnology and Bioengineering
Volume 111, Issue 8, pages 1521–1531, August 2014
How to Cite
Lian, J., Li, Y., HamediRad, M. and Zhao, H. (2014), Directed evolution of a cellodextrin transporter for improved biofuel production under anaerobic conditions in Saccharomyces cerevisiae. Biotechnol. Bioeng., 111: 1521–1531. doi: 10.1002/bit.25214
- Issue published online: 25 JUN 2014
- Article first published online: 11 MAR 2014
- Accepted manuscript online: 12 FEB 2014 06:38AM EST
- Manuscript Accepted: 3 FEB 2014
- Manuscript Revised: 6 JAN 2014
- Manuscript Received: 13 OCT 2013
- Energy Biosciences Institute
Additional supporting information may be found in the online version of this article at the publisher's web-site.
Figure S1. Vector map of the helper plasmid for CDT2 engineering, CDT-Eng-H.
Figure S2. A general scheme to engineer sugar transporters, using the colony size based screening method.
Figure S3. Characterization of the role of Q207H/F209I/N311H (HIH)mutations in cellobiose fermentation performance by creating all possible combinations, including Q207H, F209I, N311H, Q207H/F209I (HI), Q207H/N311H (HH), and F209I/N311H (IH).
Figure S4. Anaerobic cellobiose fermentation profiles of CDT2 mutants obtained in the first(HIH), second (HH), and third (HHT) round of directed evolution using the library screening conditions in rich media.
Figure S5. Mutant analysis by site-directed mutagenesis.
Figure S6. Cellobiose fermentation profiles of CDT2 on single copy plasmid (CEN/ARS Ori) and multiple copy plasmid (2µ Ori), respectively.
Figure S7. Aerobic cellobiose fermentation profiles of CDT1, CDT2, and CDT2 mutant (HHT).
Table SI. Oligonucleotides used in this study.
Table SII. Comparison of cellobiose fermentation performance of CDT1, CDT2, and CDT2 mutants obtained in the first(HIH), second(HH), and third(HHT) round of directed evolution using high cell density fermentation.
Table SIII. Saturation mutagenesis of CDT2 at the sites of Q207, N311, and I505.
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