Biosensor-based isolation of amino acid-producing Vibrio natriegens strains

The marine bacterium Vibrio natriegens has recently been demonstrated to be a promising new host for molecular biology and next generation bioprocesses. V. natriegens is a Gram-negative, non-pathogenic slight-halophilic bacterium, with a high nutrient versatility and a reported doubling time of unde...

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Autores principales:	Roberto Giuseppe Stella, Philipp Baumann, Sophia Lorke, Felix Münstermann, Astrid Wirtz, Johanna Wiechert, Jan Marienhagen, Julia Frunzke
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Transcription factor-based biosensors Metabolic engineering Amino acid production High-throughput screening Biotechnology TP248.13-248.65 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/ed0fad5b3ae54cc8a959009aaabaf611
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spelling	oai:doaj.org-article:ed0fad5b3ae54cc8a959009aaabaf6112021-11-14T04:32:54ZBiosensor-based isolation of amino acid-producing Vibrio natriegens strains2214-030110.1016/j.mec.2021.e00187https://doaj.org/article/ed0fad5b3ae54cc8a959009aaabaf6112021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2214030121000274https://doaj.org/toc/2214-0301The marine bacterium Vibrio natriegens has recently been demonstrated to be a promising new host for molecular biology and next generation bioprocesses. V. natriegens is a Gram-negative, non-pathogenic slight-halophilic bacterium, with a high nutrient versatility and a reported doubling time of under 10 min. However, V. natriegens is not an established model organism yet, and further research is required to promote its transformation into a microbial workhorse.In this work, the potential of V. natriegens as an amino acid producer was investigated. First, the transcription factor-based biosensor LysG, from Corynebacterium glutamicum, was adapted for expression in V. natriegens to facilitate the detection of positively charged amino acids. A set of different biosensor variants were constructed and characterized, using the expression of a fluorescent protein as sensor output. After random mutagenesis, one of the LysG-based sensors was used to screen for amino acid producer strains. Here, fluorescence-activated cell sorting enabled the selective sorting of highly fluorescent cells, i.e. potential producer cells. Using this approach, individual L-lysine, L-arginine and L-histidine producers could be obtained producing up to 1 mM of the effector amino acid, extracellularly. Genome sequencing of the producer strains provided insight into the amino acid production metabolism of V. natriegens.This work demonstrates the successful expression and application of transcription factor-based biosensors in V. natriegens and provides insight into the underlying physiology, forming a solid basis for further development of this promising microbe.Roberto Giuseppe StellaPhilipp BaumannSophia LorkeFelix MünstermannAstrid WirtzJohanna WiechertJan MarienhagenJulia FrunzkeElsevierarticleTranscription factor-based biosensorsMetabolic engineeringAmino acid productionHigh-throughput screeningBiotechnologyTP248.13-248.65Biology (General)QH301-705.5ENMetabolic Engineering Communications, Vol 13, Iss , Pp e00187- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Transcription factor-based biosensors Metabolic engineering Amino acid production High-throughput screening Biotechnology TP248.13-248.65 Biology (General) QH301-705.5
spellingShingle	Transcription factor-based biosensors Metabolic engineering Amino acid production High-throughput screening Biotechnology TP248.13-248.65 Biology (General) QH301-705.5 Roberto Giuseppe Stella Philipp Baumann Sophia Lorke Felix Münstermann Astrid Wirtz Johanna Wiechert Jan Marienhagen Julia Frunzke Biosensor-based isolation of amino acid-producing Vibrio natriegens strains
description	The marine bacterium Vibrio natriegens has recently been demonstrated to be a promising new host for molecular biology and next generation bioprocesses. V. natriegens is a Gram-negative, non-pathogenic slight-halophilic bacterium, with a high nutrient versatility and a reported doubling time of under 10 min. However, V. natriegens is not an established model organism yet, and further research is required to promote its transformation into a microbial workhorse.In this work, the potential of V. natriegens as an amino acid producer was investigated. First, the transcription factor-based biosensor LysG, from Corynebacterium glutamicum, was adapted for expression in V. natriegens to facilitate the detection of positively charged amino acids. A set of different biosensor variants were constructed and characterized, using the expression of a fluorescent protein as sensor output. After random mutagenesis, one of the LysG-based sensors was used to screen for amino acid producer strains. Here, fluorescence-activated cell sorting enabled the selective sorting of highly fluorescent cells, i.e. potential producer cells. Using this approach, individual L-lysine, L-arginine and L-histidine producers could be obtained producing up to 1 mM of the effector amino acid, extracellularly. Genome sequencing of the producer strains provided insight into the amino acid production metabolism of V. natriegens.This work demonstrates the successful expression and application of transcription factor-based biosensors in V. natriegens and provides insight into the underlying physiology, forming a solid basis for further development of this promising microbe.
format	article
author	Roberto Giuseppe Stella Philipp Baumann Sophia Lorke Felix Münstermann Astrid Wirtz Johanna Wiechert Jan Marienhagen Julia Frunzke
author_facet	Roberto Giuseppe Stella Philipp Baumann Sophia Lorke Felix Münstermann Astrid Wirtz Johanna Wiechert Jan Marienhagen Julia Frunzke
author_sort	Roberto Giuseppe Stella
title	Biosensor-based isolation of amino acid-producing Vibrio natriegens strains
title_short	Biosensor-based isolation of amino acid-producing Vibrio natriegens strains
title_full	Biosensor-based isolation of amino acid-producing Vibrio natriegens strains
title_fullStr	Biosensor-based isolation of amino acid-producing Vibrio natriegens strains
title_full_unstemmed	Biosensor-based isolation of amino acid-producing Vibrio natriegens strains
title_sort	biosensor-based isolation of amino acid-producing vibrio natriegens strains
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/ed0fad5b3ae54cc8a959009aaabaf611
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Biosensor-based isolation of amino acid-producing Vibrio natriegens strains

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