Enhanced brightness of bacterial luciferase by bioluminescence resonance energy transfer

Abstract Using the lux operon (luxCDABE) of bacterial bioluminescence system as an autonomous luminous reporter has been demonstrated in bacteria, plant and mammalian cells. However, applications of bacterial bioluminescence-based imaging have been limited because of its low brightness. Here, we eng...

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Autores principales:	Tomomi Kaku, Kazunori Sugiura, Tetsuyuki Entani, Kenji Osabe, Takeharu Nagai
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/80f0e38839c24c86955243024ee2d956
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spelling	oai:doaj.org-article:80f0e38839c24c86955243024ee2d9562021-12-02T17:03:49ZEnhanced brightness of bacterial luciferase by bioluminescence resonance energy transfer10.1038/s41598-021-94551-42045-2322https://doaj.org/article/80f0e38839c24c86955243024ee2d9562021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-94551-4https://doaj.org/toc/2045-2322Abstract Using the lux operon (luxCDABE) of bacterial bioluminescence system as an autonomous luminous reporter has been demonstrated in bacteria, plant and mammalian cells. However, applications of bacterial bioluminescence-based imaging have been limited because of its low brightness. Here, we engineered the bacterial luciferase (heterodimer of luxA and luxB) by fusion with Venus, a bright variant of yellow fluorescent protein, to induce bioluminescence resonance energy transfer (BRET). By using decanal as an externally added substrate, color change and ten-times enhancement of brightness was achieved in Escherichia coli when circularly permuted Venus was fused to the C-terminus of luxB. Expression of the Venus-fused luciferase in human embryonic kidney cell lines (HEK293T) or in Nicotiana benthamiana leaves together with the substrate biosynthesis-related genes (luxC, luxD and luxE) enhanced the autonomous bioluminescence. We believe the improved luciferase will forge the way towards the potential development of autobioluminescent reporter system allowing spatiotemporal imaging in live cells.Tomomi KakuKazunori SugiuraTetsuyuki EntaniKenji OsabeTakeharu NagaiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Tomomi Kaku Kazunori Sugiura Tetsuyuki Entani Kenji Osabe Takeharu Nagai Enhanced brightness of bacterial luciferase by bioluminescence resonance energy transfer
description	Abstract Using the lux operon (luxCDABE) of bacterial bioluminescence system as an autonomous luminous reporter has been demonstrated in bacteria, plant and mammalian cells. However, applications of bacterial bioluminescence-based imaging have been limited because of its low brightness. Here, we engineered the bacterial luciferase (heterodimer of luxA and luxB) by fusion with Venus, a bright variant of yellow fluorescent protein, to induce bioluminescence resonance energy transfer (BRET). By using decanal as an externally added substrate, color change and ten-times enhancement of brightness was achieved in Escherichia coli when circularly permuted Venus was fused to the C-terminus of luxB. Expression of the Venus-fused luciferase in human embryonic kidney cell lines (HEK293T) or in Nicotiana benthamiana leaves together with the substrate biosynthesis-related genes (luxC, luxD and luxE) enhanced the autonomous bioluminescence. We believe the improved luciferase will forge the way towards the potential development of autobioluminescent reporter system allowing spatiotemporal imaging in live cells.
format	article
author	Tomomi Kaku Kazunori Sugiura Tetsuyuki Entani Kenji Osabe Takeharu Nagai
author_facet	Tomomi Kaku Kazunori Sugiura Tetsuyuki Entani Kenji Osabe Takeharu Nagai
author_sort	Tomomi Kaku
title	Enhanced brightness of bacterial luciferase by bioluminescence resonance energy transfer
title_short	Enhanced brightness of bacterial luciferase by bioluminescence resonance energy transfer
title_full	Enhanced brightness of bacterial luciferase by bioluminescence resonance energy transfer
title_fullStr	Enhanced brightness of bacterial luciferase by bioluminescence resonance energy transfer
title_full_unstemmed	Enhanced brightness of bacterial luciferase by bioluminescence resonance energy transfer
title_sort	enhanced brightness of bacterial luciferase by bioluminescence resonance energy transfer
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/80f0e38839c24c86955243024ee2d956
work_keys_str_mv	AT tomomikaku enhancedbrightnessofbacterialluciferasebybioluminescenceresonanceenergytransfer AT kazunorisugiura enhancedbrightnessofbacterialluciferasebybioluminescenceresonanceenergytransfer AT tetsuyukientani enhancedbrightnessofbacterialluciferasebybioluminescenceresonanceenergytransfer AT kenjiosabe enhancedbrightnessofbacterialluciferasebybioluminescenceresonanceenergytransfer AT takeharunagai enhancedbrightnessofbacterialluciferasebybioluminescenceresonanceenergytransfer
_version_	1718381915496513536

Enhanced brightness of bacterial luciferase by bioluminescence resonance energy transfer

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