文献类型：期刊文献

英文题名：Cloning and characterization of a panel of constitutive promoters for applications in pathway engineering in Saccharomyces cerevisiae

作者：Sun, Jie[1,2];Shao, Zengyi[1];Zhao, Hua[1];Nair, Nikhil[1];Wen, Fei[1];Xu, Jian-He[2];Zhao, Huimin[1,3]

机构：[1]Univ Illinois, Dept Chem & Biomol Engn, Urbana, IL 61801 USA;[2]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[3]Univ Illinois, Dept Chem Biochem & Bioengn, Inst Genom Biol, Ctr Biophys & Computat Biol, Urbana, IL 61801 USA

年份：2012

卷号：109

期号：8

起止页码：2082

外文期刊名：BIOTECHNOLOGY AND BIOENGINEERING

收录：;EI(收录号：20122615180006);WOS:【SCI-EXPANDED(收录号:WOS:000305451400021)】；

基金：Contract grant sponsor: University of Illinois at Urbana-ChampaignContract grant sponsor: China Scholarship CouncilContract grant number: 2008674002This work was supported by the Centennial Chair Professorship in the Department of Chemical and Biomolecular Engineering at the University of Illinois at Urbana-Champaign and the China Scholarship Council (File No. 2008674002).

语种：英文

外文关键词：yeast promoters; xylanases; zeaxanthin; consolidated bioprocessing; synthetic biology

摘要：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.