@article{Yu:2022:nar/gkac476,
author = {Yu, W and Jin, K and Wu, Y and Zhang, Q and Liu, Y and Li, J and Du, G and Chen, J and Lv, X and Ledesma-Amaro, R and Liu, L},
doi = {nar/gkac476},
journal = {Nucleic Acids Research},
pages = {6587--6600},
title = {A pathway independent multi-modular ordered control system based on thermosensors and CRISPRi improves bioproduction in Bacillus subtilis},
url = {http://dx.doi.org/10.1093/nar/gkac476},
volume = {50},
year = {2022}
}

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TY  - JOUR
AB  - Dynamic regulation is an effective strategy for control of gene expression in microbial cell factories. In some pathway contexts, several metabolic modules must be controlled in a time dependent or ordered manner to maximize production, while the creation of genetic circuits with ordered regulation capacity still remains a great challenge. In this work, we develop a pathway independent and programmable system that enables multi-modular ordered control of metabolism in Bacillus subtilis. First, a series of thermosensors were created and engineered to expand their thresholds. Then we designed single-input-multi-output circuits for ordered control based on the use of thermosensors with different transition points. Meanwhile, a repression circuit was constructed by combining CRISPRi-based NOT gates. As a proof-of-concept, these genetic circuits were applied for multi-modular ordered control of 2′-fucosyllactose (2′-FL) biosynthesis, resulting in a production of 1839.7 mg/l in shake flask, which is 5.16-times that of the parental strain. In a 5-l bioreactor, the 2′-FL titer reached 28.2 g/l with down-regulation of autolysis. Taken together, this work provides programmable and versatile thermosensitive genetic toolkits for dynamic regulation in B. subtilis and a multi-modular ordered control framework that can be used to improve metabolic modules in other chassis cells and for other compounds.
AU  - Yu,W
AU  - Jin,K
AU  - Wu,Y
AU  - Zhang,Q
AU  - Liu,Y
AU  - Li,J
AU  - Du,G
AU  - Chen,J
AU  - Lv,X
AU  - Ledesma-Amaro,R
AU  - Liu,L
DO  - nar/gkac476
EP  - 6600
PY  - 2022///
SN  - 0305-1048
SP  - 6587
TI  - A pathway independent multi-modular ordered control system based on thermosensors and CRISPRi improves bioproduction in Bacillus subtilis
T2  - Nucleic Acids Research
UR  - http://dx.doi.org/10.1093/nar/gkac476
UR  - http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000807082600001&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
UR  - https://academic.oup.com/nar/article/50/11/6587/6603658
UR  - http://hdl.handle.net/10044/1/98862
VL  - 50
ER  - 

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