OptoLacI: optogenetically engineered lactose operon repressor LacI responsive to light instead of IPTG.

Publisher: Oxford University Press Country of Publication: England NLM ID: 0411011 Publication Model: Print Cited Medium: Internet ISSN: 1362-4962 (Electronic) Linking ISSN: 03051048 NLM ISO Abbreviation: Nucleic Acids Res Subsets: MEDLINE

Publication: 1992- : Oxford : Oxford University Press
Original Publication: London, Information Retrieval ltd.

Isopropyl Thiogalactoside*/pharmacology ; Lac Repressors*/metabolism ; Lac Repressors*/genetics ; Escherichia coli*/genetics ; Escherichia coli*/metabolism ; Escherichia coli*/radiation effects ; Light* ; Optogenetics*/methods ; Lac Operon*; Gene Expression Regulation, Bacterial/radiation effects ; Escherichia coli Proteins/genetics ; Escherichia coli Proteins/metabolism ; Protein Engineering/methods ; Avena/genetics ; Avena/metabolism ; Avena/radiation effects

Optogenetics' advancement has made light induction attractive for controlling biological processes due to its advantages of fine-tunability, reversibility, and low toxicity. The lactose operon induction system, commonly used in Escherichia coli, relies on the binding of lactose or isopropyl β-d-1-thiogalactopyranoside (IPTG) to the lactose repressor protein LacI, playing a pivotal role in controlling the lactose operon. Here, we harnessed the light-responsive light-oxygen-voltage 2 (LOV2) domain from Avena sativa phototropin 1 as a tool for light control and engineered LacI into two light-responsive variants, OptoLacIL and OptoLacID. These variants exhibit direct responsiveness to light and darkness, respectively, eliminating the need for IPTG. Building upon OptoLacI, we constructed two light-controlled E. coli gene expression systems, OptoE.coliLight system and OptoE.coliDark system. These systems enable bifunctional gene expression regulation in E. coli through light manipulation and show superior controllability compared to IPTG-induced systems. We applied the OptoE.coliDark system to protein production and metabolic flux control. Protein production levels are comparable to those induced by IPTG. Notably, the titers of dark-induced production of 1,3-propanediol (1,3-PDO) and ergothioneine exceeded 110% and 60% of those induced by IPTG, respectively. The development of OptoLacI will contribute to the advancement of the field of optogenetic protein engineering, holding substantial potential applications across various fields.
(© The Author(s) 2024. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Nat Commun. 2020 May 8;11(1):2262. (PMID: 32385264)
Acta Biochim Biophys Sin (Shanghai). 2021 Apr 15;53(5):620-627. (PMID: 33764372)
Nutr Res Rev. 2020 Dec;33(2):190-217. (PMID: 32051057)
ACS Synth Biol. 2022 Oct 21;11(10):3354-3367. (PMID: 35998606)
J Mol Biol. 1961 Jun;3:318-56. (PMID: 13718526)
Science. 1996 Mar 1;271(5253):1247-54. (PMID: 8638105)
Curr Opin Cell Biol. 2020 Oct;66:1-10. (PMID: 32371345)
Bioresour Bioprocess. 2022 Aug 13;9(1):82. (PMID: 38647602)
Nat Commun. 2018 Nov 21;9(1):4917. (PMID: 30464241)
Biochemistry. 2007 Dec 11;46(49):14001-9. (PMID: 18001137)
EMBO J. 1990 Apr;9(4):973-9. (PMID: 2182324)
Curr Opin Physiol. 2020 Oct;17:187-196. (PMID: 33184610)
J Mol Biol. 2021 Jul 23;433(15):167107. (PMID: 34146595)
Nat Commun. 2022 Aug 2;13(1):4491. (PMID: 35918338)
Biotechnol Bioeng. 2020 May;117(5):1513-1524. (PMID: 32022248)
J Agric Food Chem. 2023 Jan 11;71(1):671-679. (PMID: 36571834)
Microb Biotechnol. 2015 Mar;8(2):253-65. (PMID: 25488698)
Cell Calcium. 2018 Nov;75:79-88. (PMID: 30199756)
Org Lett. 2014 Oct 17;16(20):5382-5. (PMID: 25275953)
Biotechnol J. 2022 Sep;17(9):e2200129. (PMID: 35661423)
Science. 1995 Jun 23;268(5218):1721-7. (PMID: 7792597)
Nat Protoc. 2019 Jun;14(6):1863-1883. (PMID: 31076662)
J Mol Biol. 2001 Oct 12;313(1):99-109. (PMID: 11601849)
J Biol Chem. 1993 Aug 15;268(23):17613-22. (PMID: 8349640)
Nat Chem Biol. 2021 Jan;17(1):71-79. (PMID: 32895498)
Elife. 2021 Feb 23;10:. (PMID: 33620312)
Nucleic Acids Res. 2020 Apr 6;48(6):e33. (PMID: 31989175)
Science. 2011 Sep 2;333(6047):1307-11. (PMID: 21885784)
Bioresour Bioprocess. 2022 May 18;9(1):56. (PMID: 38647747)
Nucleic Acids Res. 2016 Aug 19;44(14):6994-7005. (PMID: 27353329)
Microb Cell Fact. 2013 Jul 08;12:67. (PMID: 23834731)
Science. 2003 Sep 12;301(5639):1541-4. (PMID: 12970567)
Sci Adv. 2022 Feb 11;8(6):eabl5166. (PMID: 35148182)
Biochemistry. 2007 Mar 20;46(11):3129-37. (PMID: 17311415)
J Mol Biol. 2011 Jan 14;405(2):315-24. (PMID: 21035461)
Cell. 2007 Jul 27;130(2):363-72. (PMID: 17662949)
Nature. 2018 Mar 29;555(7698):683-687. (PMID: 29562237)
Bioresour Technol. 2018 Nov;267:599-607. (PMID: 30056370)
Nat Commun. 2021 Jun 23;12(1):3859. (PMID: 34162879)
Biotechnol J. 2023 Sep;18(9):e2300027. (PMID: 37265188)
Trends Genet. 2022 Dec;38(12):1253-1270. (PMID: 35738948)
Nat Methods. 2012 Jul;9(7):671-5. (PMID: 22930834)
Nat Protoc. 2009;4(2):206-23. (PMID: 19180090)
ACS Synth Biol. 2023 Nov 17;12(11):3328-3339. (PMID: 37885173)
Nat Methods. 2009 May;6(5):343-5. (PMID: 19363495)
Nat Chem Biol. 2021 Jul;17(7):817-827. (PMID: 33903769)
Cell Res. 2016 Jul;26(7):854-7. (PMID: 27311594)

2020YFA0908300 National Key Research and Development Program of China; TSBICIP-CXRC-028 Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project; 22HHSWSS00002 Major Project of Haihe Laboratory of Synthetic Biology

367-93-1 (Isopropyl Thiogalactoside)
0 (Lac Repressors)
0 (Escherichia coli Proteins)
0 (LacI protein, E coli)

Date Created: 20240611 Date Completed: 20240721 Latest Revision: 20240821

20240821

PMC11260447

10.1093/nar/gkae479

38860425