Journal
METABOLIC ENGINEERING
Volume 14, Issue 4, Pages 401-411Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2012.03.004
Keywords
Sugar transport; Directed evolution; Yeast; Pentose metabolism; Lignocellulose conversion
Categories
Funding
- Camille and Henry Dreyfus New Faculty Award
- National Science Foundation Graduate Research Fellowship
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Traditional metabolic pathway engineering rarely considers the influence of molecular transport. Here, we describe the directed evolution of two heterologous transporters, Candida intermedia GXS1 and Scheffersomyces stipins XUT3. Growth rate on xylose was improved up to 70% by mutant transporter expression. Most mutants were found to exhibit vastly improved V-max values and display an increase in high cell density sugar consumption rates. Mixed glucose and xylose fermentations reveal that mutant transporters can alter the diauxic shift dynamics and the simultaneous sugar utilization capacity of the host strain. Analysis of mutations highlights several important residues influencing transporter function including point mutations at F40 of C. intermedia GXS1 and at E538 of S. stipitis XUT3. This work is the first to demonstrate that molecular transporter proteins can be improved for biotechnological applications through directed evolution in yeast. (C)) 2012 Elsevier Inc. All rights reserved.
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