BioChem - DOS

Biosensor-Based Evolution and Elucidation of a Biosynthetic Pathway in Escherichia coli

ACS Synthetic biology

Liu, Yongfei; Zhuang, Yinyin; Ding, Dongqin; Xu, Yiran; Sun, Jibin; Zhang, Dawei

<P>The successful evolution of metabolite-producing microbes requires a high-throughput screening method to obtain the desired properties within a short time. In this study, we developed a transcription-factor-driven device that combines a metabolite-responsive element and a selection module. This device was able to specifically sense intracellular <SMALL>l</SMALL>-phenylalanine (<SMALL>l</SMALL>-Phe) and convert this signal into an observable phenotype. Applying this device, we successfully improved <SMALL>l</SMALL>-Phe production by screening hyperproducing phenotypes from a ribonucleotide binding site library and a random mutagenesis library. In addition, several site mutations introduced by random mutagenesis were identified and elucidated to facilitate the improvement of <SMALL>l</SMALL>-Phe production. Our results present a paradigm for screening of compounds that are not easily observable to raise the yield of targeted compounds from a large candidate library. This approach may guide further applications in rewiring metabolic circuits and facilitate the directed evolution of recombinant strains.</P><P><B>Graphic Abstract</B><BR><IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/asbcd6/2017/asbcd6.2017.6.issue-5/acssynbio.6b00328/production/images/medium/sb-2016-00328e_0009.gif'></P><P><A href='http://pubs.acs.org/doi/suppl/10.1021/sb6b00328'>ACS Electronic Supporting Info</A></P>

2017

American Chemical Society

2161-5063

6

5

pp.837-848

10.1021/acssynbio.6b00328.s001

http://click.ndsl.kr/servlet/OpenAPIDetailView?keyValue=05787966&target=NART&cn=NART77881067

https://doi.org/10.1021/acssynbio.6b00328.s001

biosensor; transcription factor; l-phenylalanine; high-throughput screening; TyrR