Harnessing β-estradiol inducible expression system to overproduce nervonic acid in Saccharomyces cerevisiae |
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| Institution: | 1. School of Life Sciences, Ludong University, Yantai 264025, China;2. School of Agriculture, Ludong University, Yantai 264025, China;1. School of Ecology and Environment, Zhengzhou University, Zhengzhou 450001, China;2. School of Chemical Engineering, Zhengzhou University, Zhengzhou 450001, China;3. Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008, China;1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai Institute of Biomanufacturing Technology and Collaborative Innovation Center, Shanghai 200237, China;2. Department of Biotechnology, Changshu Institute of Technology, Jiangsu, China;1. School of Food and Biological Engineering, Jiangsu University, 301 Xuefu Road, Zhenjiang, 212013, China;2. Department of Biosystems Engineering and Soil Science, The University of Tennessee, 2506 E.J. Chapman Drive, Knoxville, TN, 37996, USA |
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| Abstract: | As a very long chain monounsaturated fatty acid, nervonic acid (NA) holds great application potential for brain diseases. 3-ketoacyl-CoA synthase (KCS) with substrate specificity plays a key role in the carbon chain elongation cycle for NA biosynthesis. A tunable and sensitive single-gene expression regulation is necessary to overproduce NA in yeast cell. Herein, β-estradiol inducible expression system (EIES) was employed to augment NA biosynthesis in Saccharomyces cerevisiae. EIES consisted of two recombinant vectors: pRS416-PGal1-GFP-TCYC1 and YEplac112-PTEF1-ZEV-TCYC1. The following was the optimized induction conditions of EIES in SD medium: 0.1 μM β-estradiol, 15 min of heat induction at 33 ℃ every 8 h within first 32 h. Using EIES to enhance KCS and ELOVL1 expression, NA contents in the recombinants increased by 332 % and 101 % respectively, compared with the control strain using strong PGK promoter. NA levels were further improved by knockout of elo2 in the recombinants expressing KCS or ELOVL1. This study suggested that EIES could be a promising tool for enhancing the biosynthesis of desired metabolites. |
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| Keywords: | β-estradiol inducible Expression system Nervonic acid 3-ketoacyl-CoA synthase Heat induction |
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