Strategic single point mutation yields a solvent- and salt-stable transaminase from Virgibacillus sp. in soluble form

Abstract A new transaminase (VbTA) was identified from the genome of the halotolerant marine bacterium Virgibacillus 21D. Following heterologous expression in Escherichia coli, it was located entirely in the insoluble fraction. After a single mutation, identified via sequence homology analyses, the...

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Autores principales:	Benedetta Guidi, Matteo Planchestainer, Martina Letizia Contente, Tommaso Laurenzi, Ivano Eberini, Louise J. Gourlay, Diego Romano, Francesca Paradisi, Francesco Molinari
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Virgibacillus Mutation T215F Hydration Free Energy Deep Hypersaline Anoxic Basins Acceptor Amino Medicine R Science Q
Acceso en línea:	https://doaj.org/article/77bae478128847fb89ddb990cced95fd
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spelling	oai:doaj.org-article:77bae478128847fb89ddb990cced95fd2021-12-02T15:09:11ZStrategic single point mutation yields a solvent- and salt-stable transaminase from Virgibacillus sp. in soluble form10.1038/s41598-018-34434-32045-2322https://doaj.org/article/77bae478128847fb89ddb990cced95fd2018-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-34434-3https://doaj.org/toc/2045-2322Abstract A new transaminase (VbTA) was identified from the genome of the halotolerant marine bacterium Virgibacillus 21D. Following heterologous expression in Escherichia coli, it was located entirely in the insoluble fraction. After a single mutation, identified via sequence homology analyses, the VbTA T16F mutant was successfully expressed in soluble form and characterised. VbTA T16F showed high stability towards polar organic solvents and salt exposure, accepting mainly hydrophobic aromatic amine and carbonyl substrates. The 2.0 Å resolution crystal structure of VbTA T16F is here reported, and together with computational calculations, revealed that this mutation is crucial for correct dimerisation and thus correct folding, leading to soluble protein expression.Benedetta GuidiMatteo PlanchestainerMartina Letizia ContenteTommaso LaurenziIvano EberiniLouise J. GourlayDiego RomanoFrancesca ParadisiFrancesco MolinariNature PortfolioarticleVirgibacillusMutation T215FHydration Free EnergyDeep Hypersaline Anoxic BasinsAcceptor AminoMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-11 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Virgibacillus Mutation T215F Hydration Free Energy Deep Hypersaline Anoxic Basins Acceptor Amino Medicine R Science Q
spellingShingle	Virgibacillus Mutation T215F Hydration Free Energy Deep Hypersaline Anoxic Basins Acceptor Amino Medicine R Science Q Benedetta Guidi Matteo Planchestainer Martina Letizia Contente Tommaso Laurenzi Ivano Eberini Louise J. Gourlay Diego Romano Francesca Paradisi Francesco Molinari Strategic single point mutation yields a solvent- and salt-stable transaminase from Virgibacillus sp. in soluble form
description	Abstract A new transaminase (VbTA) was identified from the genome of the halotolerant marine bacterium Virgibacillus 21D. Following heterologous expression in Escherichia coli, it was located entirely in the insoluble fraction. After a single mutation, identified via sequence homology analyses, the VbTA T16F mutant was successfully expressed in soluble form and characterised. VbTA T16F showed high stability towards polar organic solvents and salt exposure, accepting mainly hydrophobic aromatic amine and carbonyl substrates. The 2.0 Å resolution crystal structure of VbTA T16F is here reported, and together with computational calculations, revealed that this mutation is crucial for correct dimerisation and thus correct folding, leading to soluble protein expression.
format	article
author	Benedetta Guidi Matteo Planchestainer Martina Letizia Contente Tommaso Laurenzi Ivano Eberini Louise J. Gourlay Diego Romano Francesca Paradisi Francesco Molinari
author_facet	Benedetta Guidi Matteo Planchestainer Martina Letizia Contente Tommaso Laurenzi Ivano Eberini Louise J. Gourlay Diego Romano Francesca Paradisi Francesco Molinari
author_sort	Benedetta Guidi
title	Strategic single point mutation yields a solvent- and salt-stable transaminase from Virgibacillus sp. in soluble form
title_short	Strategic single point mutation yields a solvent- and salt-stable transaminase from Virgibacillus sp. in soluble form
title_full	Strategic single point mutation yields a solvent- and salt-stable transaminase from Virgibacillus sp. in soluble form
title_fullStr	Strategic single point mutation yields a solvent- and salt-stable transaminase from Virgibacillus sp. in soluble form
title_full_unstemmed	Strategic single point mutation yields a solvent- and salt-stable transaminase from Virgibacillus sp. in soluble form
title_sort	strategic single point mutation yields a solvent- and salt-stable transaminase from virgibacillus sp. in soluble form
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/77bae478128847fb89ddb990cced95fd
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Strategic single point mutation yields a solvent- and salt-stable transaminase from Virgibacillus sp. in soluble form

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