New insights into the molecular mechanism behind mannitol and erythritol fructosylation by β-fructofuranosidase from Schwanniomyces occidentalis

Abstract The β-fructofuranosidase from Schwanniomyces occidentalis (Ffase) is a useful biotechnological tool for the fructosylation of different acceptors to produce fructooligosaccharides (FOS) and fructo-conjugates. In this work, the structural determinants of Ffase involved in the transfructosyla...

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Autores principales:	David Rodrigo-Frutos, Elena Jiménez-Ortega, David Piedrabuena, Mercedes Ramírez-Escudero, Noa Míguez, Francisco J. Plou, Julia Sanz-Aparicio, María Fernández-Lobato
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8e2197f6a80a4f17a0a1b8b746479e11
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spelling	oai:doaj.org-article:8e2197f6a80a4f17a0a1b8b746479e112021-12-02T18:17:42ZNew insights into the molecular mechanism behind mannitol and erythritol fructosylation by β-fructofuranosidase from Schwanniomyces occidentalis10.1038/s41598-021-86568-62045-2322https://doaj.org/article/8e2197f6a80a4f17a0a1b8b746479e112021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-86568-6https://doaj.org/toc/2045-2322Abstract The β-fructofuranosidase from Schwanniomyces occidentalis (Ffase) is a useful biotechnological tool for the fructosylation of different acceptors to produce fructooligosaccharides (FOS) and fructo-conjugates. In this work, the structural determinants of Ffase involved in the transfructosylating reaction of the alditols mannitol and erythritol have been studied in detail. Complexes with fructosyl-erythritol or sucrose were analyzed by crystallography and the effect of mutational changes in positions Gln-176, Gln-228, and Asn-254 studied to explore their role in modulating this biocatalytic process. Interestingly, N254T variant enhanced the wild-type protein production of fructosyl-erythritol and FOS by $$\sim$$ ∼ 30% and 48%, respectively. Moreover, it produced neokestose, which represented $$\sim$$ ∼ 27% of total FOS, and yielded 31.8 g l−1 blastose by using glucose as exclusive fructosyl-acceptor. Noteworthy, N254D and Q176E replacements turned the specificity of Ffase transferase activity towards the synthesis of the fructosylated polyols at the expense of FOS production, but without increasing the total reaction efficiency. The results presented here highlight the relevance of the pair Gln-228/Asn-254 for Ffase donor-sucrose binding and opens new windows of opportunity for optimizing the generation of fructosyl-derivatives by this enzyme enhancing its biotechnological applicability.David Rodrigo-FrutosElena Jiménez-OrtegaDavid PiedrabuenaMercedes Ramírez-EscuderoNoa MíguezFrancisco J. PlouJulia Sanz-AparicioMaría Fernández-LobatoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q David Rodrigo-Frutos Elena Jiménez-Ortega David Piedrabuena Mercedes Ramírez-Escudero Noa Míguez Francisco J. Plou Julia Sanz-Aparicio María Fernández-Lobato New insights into the molecular mechanism behind mannitol and erythritol fructosylation by β-fructofuranosidase from Schwanniomyces occidentalis
description	Abstract The β-fructofuranosidase from Schwanniomyces occidentalis (Ffase) is a useful biotechnological tool for the fructosylation of different acceptors to produce fructooligosaccharides (FOS) and fructo-conjugates. In this work, the structural determinants of Ffase involved in the transfructosylating reaction of the alditols mannitol and erythritol have been studied in detail. Complexes with fructosyl-erythritol or sucrose were analyzed by crystallography and the effect of mutational changes in positions Gln-176, Gln-228, and Asn-254 studied to explore their role in modulating this biocatalytic process. Interestingly, N254T variant enhanced the wild-type protein production of fructosyl-erythritol and FOS by $$\sim$$ ∼ 30% and 48%, respectively. Moreover, it produced neokestose, which represented $$\sim$$ ∼ 27% of total FOS, and yielded 31.8 g l−1 blastose by using glucose as exclusive fructosyl-acceptor. Noteworthy, N254D and Q176E replacements turned the specificity of Ffase transferase activity towards the synthesis of the fructosylated polyols at the expense of FOS production, but without increasing the total reaction efficiency. The results presented here highlight the relevance of the pair Gln-228/Asn-254 for Ffase donor-sucrose binding and opens new windows of opportunity for optimizing the generation of fructosyl-derivatives by this enzyme enhancing its biotechnological applicability.
format	article
author	David Rodrigo-Frutos Elena Jiménez-Ortega David Piedrabuena Mercedes Ramírez-Escudero Noa Míguez Francisco J. Plou Julia Sanz-Aparicio María Fernández-Lobato
author_facet	David Rodrigo-Frutos Elena Jiménez-Ortega David Piedrabuena Mercedes Ramírez-Escudero Noa Míguez Francisco J. Plou Julia Sanz-Aparicio María Fernández-Lobato
author_sort	David Rodrigo-Frutos
title	New insights into the molecular mechanism behind mannitol and erythritol fructosylation by β-fructofuranosidase from Schwanniomyces occidentalis
title_short	New insights into the molecular mechanism behind mannitol and erythritol fructosylation by β-fructofuranosidase from Schwanniomyces occidentalis
title_full	New insights into the molecular mechanism behind mannitol and erythritol fructosylation by β-fructofuranosidase from Schwanniomyces occidentalis
title_fullStr	New insights into the molecular mechanism behind mannitol and erythritol fructosylation by β-fructofuranosidase from Schwanniomyces occidentalis
title_full_unstemmed	New insights into the molecular mechanism behind mannitol and erythritol fructosylation by β-fructofuranosidase from Schwanniomyces occidentalis
title_sort	new insights into the molecular mechanism behind mannitol and erythritol fructosylation by β-fructofuranosidase from schwanniomyces occidentalis
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/8e2197f6a80a4f17a0a1b8b746479e11
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New insights into the molecular mechanism behind mannitol and erythritol fructosylation by β-fructofuranosidase from Schwanniomyces occidentalis

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