title: Fine-Tuning of the Fatty Acid Pathway by Synthetic Antisense RNA for Enhanced (2S)-Naringenin Production from l-Tyrosine in Escherichia coli.
abstract: Malonyl coenzyme A (malonyl-CoA) is an important precursor for the synthesis of natural products, such as polyketides and flavonoids. The majority of this cofactor often is consumed for producing fatty acids and phospholipids, leaving only a small amount of cellular malonyl-CoA available for producing the target compound. The tuning of malonyl-CoA into heterologous pathways yields significant phenotypic effects, such as growth retardation and even cell death. In this study, fine-tuning of the fatty acid pathway in Escherichia coli with antisense RNA (asRNA) to balance the demands on malonyl-CoA for target-product synthesis and cell health was proposed. To establish an efficient asRNA system, the relationship between sequence and function for asRNA was explored. It was demonstrated that the gene-silencing effect of asRNA could be tuned by directing asRNA to different positions in the 5'-UTR (untranslated region) of the target gene. Based on this principle, the activity of asRNA was quantitatively tailored to balance the need for malonyl-CoA in cell growth and the production of the main flavonoid precursor, (2S)-naringenin. Appropriate inhibitory efficiency of the anti-fabB/fabF asRNA improved the production titer by 431% (391 mg/liter). Therefore, the strategy presented in this study provided a useful tool for the fine-tuning of endogenous gene expression in bacteria.
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China Synergetic Innovation Center of Food Safety and Nutrition, Wuxi, Jiangsu, China.
authors.forename: Junjun
authors.initials: J
authors.lastname: Wu
authors.affiliation: Donald Danforth Plant Science Center, St. Louis, Missouri, USA.
authors.forename: Oliver
authors.initials: O
authors.lastname: Yu
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China Synergetic Innovation Center of Food Safety and Nutrition, Wuxi, Jiangsu, China.
authors.forename: Guocheng
authors.initials: G
authors.lastname: Du
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China Synergetic Innovation Center of Food Safety and Nutrition, Wuxi, Jiangsu, China zhoujw1982@jiangnan.edu.cn jchen@jiangnan.edu.cn.
authors.forename: Jingwen
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, Jiangsu, China Synergetic Innovation Center of Food Safety and Nutrition, Wuxi, Jiangsu, China zhoujw1982@jiangnan.edu.cn jchen@jiangnan.edu.cn.
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
mesh_terms: D003600:Cytosol / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D044950:Flavanones / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D008316:Malonyl Coenzyme A / Q000378:metabolism; D060847:Metabolic Engineering / Q000379:methods*; D010743:Phospholipids / Q000032:analysis; D016372:RNA, Antisense / Q000235:genetics / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism*
chemical_list: D044950:Flavanones; D010743:Phospholipids; D016372:RNA, Antisense; D014443:Tyrosine; D008316:Malonyl Coenzyme A; C005273:naringenin