Journal
BIOTECHNOLOGY AND BIOENGINEERING
Volume 109, Issue 8, Pages 2082-2092Publisher
WILEY
DOI: 10.1002/bit.24481
Keywords
yeast promoters; xylanases; zeaxanthin; consolidated bioprocessing; synthetic biology
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Funding
- University of Illinois at Urbana-Champaign
- China Scholarship Council [2008674002]
- Centennial Chair Professorship in the Department of Chemical and Biomolecular Engineering at the University of Illinois at Urbana-Champaign
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Saccharomyces cerevisiae is an important platform organism for synthesis of chemicals and fuels. However, the promoters used in most pathway engineering studies in S. cerevisiae have not been characterized and compared in parallel under multiple conditions that are routinely operated in laboratory and the number of known promoters is rather limited for the construction of large biochemical pathways. Here a total of 14 constitutive promoters from S. cerevisiae were cloned and characterized using a green fluorescent protein (GFP) as a reporter in a 2?mu vector pRS426, under varying glucose and oxygen concentrations. The strengths of these promoters varied no more than sixfold in the mean fluorescence intensity of GFP, with promoter TEF1p being the strongest and promoter PGI1p the weakest. As an example of application for these promoters in metabolic engineering, the genes involved in xylan degradation and zeaxanthin biosynthesis were subsequently cloned under the control of promoters with medium to high strength and assembled into a single pathway. The corresponding construct was transformed to a S. cerevisiae strain integrated with a D-xylose utilizing pathway. The resulting strain produced zeaxanthin with a titer of 0.74?+/-?0.02?mg/L directly from birchwood xylan. Biotechnol. Bioeng. 2012; 109:20822092. (c) 2012 Wiley Periodicals, Inc.
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