TY - JOUR
T1 - Optogenetic control of the lac operon for bacterial chemical and protein production
AU - Lalwani, Makoto A.
AU - Ip, Samantha S.
AU - Carrasco-López, César
AU - Day, Catherine
AU - Zhao, Evan M.
AU - Kawabe, Hinako
AU - Avalos, José L.
N1 - Funding Information:
We thank M. Brynildsen for strain MG1655 ΔlacI::FRT-KanR-FRT, A. Möglich for plasmids pDusk and pDawn, J. Keasling for plasmid pMevT and J. Liao for plasmids pSA65 and pSA69. We are very grateful to W. Mok and M. Brynildsen for advice and troubleshooting regarding E. coli protocols. We thank C. DeCoste, K. Rittenbach and the Princeton Molecular Biology Flow Cytometry Resource Center for assistance with flow cytometry experiments. J.L.A. is supported by the US Department of Energy, Office of Science, Office of Biological and Environmental Research award no. DE-SC0019363, the NSF CAREER Award CBET-1751840, The Pew Charitable Trusts, The Eric and Wendy Schmidt Transformative Technology Fund Award and the Camille Dreyfus Teacher-Scholar Award.
Publisher Copyright:
© 2020, The Author(s), under exclusive licence to Springer Nature America, Inc.
PY - 2021/1
Y1 - 2021/1
N2 - Control of the lac operon with isopropyl β-d-1-thiogalactopyranoside (IPTG) has been used to regulate gene expression in Escherichia coli for countless applications, including metabolic engineering and recombinant protein production. However, optogenetics offers unique capabilities, such as easy tunability, reversibility, dynamic induction strength and spatial control, that are difficult to obtain with chemical inducers. We have developed a series of circuits for optogenetic regulation of the lac operon, which we call OptoLAC, to control gene expression from various IPTG-inducible promoters using only blue light. Applying them to metabolic engineering improves mevalonate and isobutanol production by 24% and 27% respectively, compared to IPTG induction, in light-controlled fermentations scalable to at least two-litre bioreactors. Furthermore, OptoLAC circuits enable control of recombinant protein production, reaching yields comparable to IPTG induction but with easier tunability of expression. OptoLAC circuits are potentially useful to confer light control over other cell functions originally designed to be IPTG-inducible. [Figure not available: see fulltext.]
AB - Control of the lac operon with isopropyl β-d-1-thiogalactopyranoside (IPTG) has been used to regulate gene expression in Escherichia coli for countless applications, including metabolic engineering and recombinant protein production. However, optogenetics offers unique capabilities, such as easy tunability, reversibility, dynamic induction strength and spatial control, that are difficult to obtain with chemical inducers. We have developed a series of circuits for optogenetic regulation of the lac operon, which we call OptoLAC, to control gene expression from various IPTG-inducible promoters using only blue light. Applying them to metabolic engineering improves mevalonate and isobutanol production by 24% and 27% respectively, compared to IPTG induction, in light-controlled fermentations scalable to at least two-litre bioreactors. Furthermore, OptoLAC circuits enable control of recombinant protein production, reaching yields comparable to IPTG induction but with easier tunability of expression. OptoLAC circuits are potentially useful to confer light control over other cell functions originally designed to be IPTG-inducible. [Figure not available: see fulltext.]
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U2 - 10.1038/s41589-020-0639-1
DO - 10.1038/s41589-020-0639-1
M3 - Article
C2 - 32895498
AN - SCOPUS:85090310024
SN - 1552-4450
VL - 17
SP - 71
EP - 79
JO - Nature Chemical Biology
JF - Nature Chemical Biology
IS - 1
ER -