文献类型：期刊文献

英文题名：Naringenin-responsive riboswitch-based fluorescent biosensor module for Escherichia coli co-cultures

作者：Xiu, Yu[1,2,3];Jang, Sungho[4];Jones, J. Andrew[3,5];Zill, Nicholas A.[3];Linhardt, Robert J.[3];Yuan, Qipeng[1];Jung, Gyoo Yeol[4,6];Koffas, Mattheos A. G.[3]

第一作者：Xiu, Yu

通讯作者：Koffas, MAG[1]

机构：[1]Beijing Univ Chem Technol, State Key Lab Chem Resource Engn, Beijing, Peoples R China;[2]Beijing Union Univ, Beijing Key Lab Bioact Subst & Funct Food, Beijing, Peoples R China;[3]Rensselaer Polytech Inst, Dept Chem & Biol Engn, Ctr Biotechnol & Interdisciplinary Studies, Troy, NY 12180 USA;[4]Pohang Univ Sci & Technol, Dept Chem Engn, Pohang, Gyeongbuk, South Korea;[5]Hamilton Coll, Dept Chem, Clinton, NY 13323 USA;[6]Pohang Univ Sci & Technol, Sch Interdisciplinary Biosci & Bioengn, Pohang, Gyeongbuk, South Korea

第一机构：Beijing Univ Chem Technol, State Key Lab Chem Resource Engn, Beijing, Peoples R China

通讯机构：[1]corresponding author), Rensselaer Polytech Inst, Dept Chem & Biol Engn, Ctr Biotechnol & Interdisciplinary Studies, Troy, NY 12180 USA.

年份：2017

卷号：114

期号：10

起止页码：2235-2244

外文期刊名：BIOTECHNOLOGY AND BIOENGINEERING

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000408777100008)】；

基金：Contract grant sponsor: National Research Foundation of Korea; Contract grant numbers: NRF-2015R1A2A1A10056126; NRF-2016K1A1A2912829; Contract grant sponsor: National Natural Science Foundation of China; Contract grant number: 21376017

语种：英文

外文关键词：riboswitch; co-culture; RNA aptamer; naringenin; flow cytometry; flavonoids; biosensor design

摘要：The ability to design and construct combinatorial synthetic metabolic pathways has far exceeded our capacity for efficient screening and selection of the resulting microbial strains. The need for high-throughput rapid screening techniques is of upmost importance for the future of synthetic biology and metabolic engineering. Here we describe the development of an RNA riboswitch-based biosensor module with dual fluorescent reporters, and demonstrate a high-throughput flow cytometry-based screening method for identification of naringenin over producing Escherichia coli strains in co-culture. Our efforts helped identify a number of key operating parameters that affect biosensor performance, including the selection of promoter and linker elements within the sensor-actuator domain, and the effect of host strain, fermentation time, and growth medium on sensor dynamic range. The resulting biosensor demonstrates a high correlation between specific fluorescence of the biosensor strain and naringenin titer produced by the second member of the synthetic co-culture system. This technique represents a novel application for synthetic microbial co-cultures and can be expanded from naringenin to any metabolite if a suitable riboswitch is identified. The co-culture technique presented here can be applied to a variety of target metabolites in combination with the SELEX approach for aptamer design. Due to the compartmentalization of the two genetic constructs responsible for production and detection into separate cells and application as independent modules of a synthetic microbial co-culture we have subsequently reduced the need for re-optimization of the producer module when the biosensor is replaced or removed. Biotechnol. Bioeng. 2017;114: 2235-2244. (c) 2017 Wiley Periodicals, Inc.