Article
Cloning and characterization of a panel of constitutive promoters for applications in pathway engineering in Saccharomyces cerevisiae
Biotechnology and Bioengineering
(2012)
Abstract
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. cerevisiaewere cloned and characterized using a green fluorescent protein (GFP) as a reporter in a 2 µ 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.
Keywords
- yeast promoters,
- xylanases,
- zeaxanthin,
- consolidated bioproccessing,
- synthetic biology
Disciplines
Publication Date
2012
DOI
10.1002/bit.24481
Publisher Statement
This is the peer reviewed version of the following article: Sun, Jie, Zengyi Shao, Hua Zhao, Nikhil Nair, Fei Wen, Jian‐He Xu, and Huimin Zhao. "Cloning and characterization of a panel of constitutive promoters for applications in pathway engineering in Saccharomyces cerevisiae." Biotechnology and bioengineering 109, no. 8 (2012): 2082-2092., which has been published in final form at DOI: 10.1002/bit.24481. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.
Citation Information
Jie Sun, Zengyi Shao, Hua Zhao, Nikhil Nair, et al.. "Cloning and characterization of a panel of constitutive promoters for applications in pathway engineering in Saccharomyces cerevisiae" Biotechnology and Bioengineering Vol. 109 Iss. 8 (2012) p. 2082 - 2092 Available at: http://works.bepress.com/zengyi_shao/9/