Glucansucrase (mutant) enzymes from Lactobacillus reuteri 180 efficiently transglucosylate Stevia component rebaudioside A, resulting in a superior taste

Abstract Steviol glycosides from the leaves of the plant Stevia rebaudiana are high-potency natural sweeteners but suffer from a lingering bitterness. The Lactobacillus reuteri 180 wild-type glucansucrase Gtf180-ΔN, and in particular its Q1140E-mutant, efficiently α-glucosylated rebaudioside A (RebA...

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Autores principales:	Evelien M. te Poele, Tim Devlamynck, Manuel Jäger, Gerrit J. Gerwig, Davy Van de Walle, Koen Dewettinck, Anna K. H. Hirsch, Johannis P. Kamerling, Wim Soetaert, Lubbert Dijkhuizen
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/5e79b1d8bf0d4f5a884762ce3f6ab06c
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spelling	oai:doaj.org-article:5e79b1d8bf0d4f5a884762ce3f6ab06c2021-12-02T15:08:48ZGlucansucrase (mutant) enzymes from Lactobacillus reuteri 180 efficiently transglucosylate Stevia component rebaudioside A, resulting in a superior taste10.1038/s41598-018-19622-52045-2322https://doaj.org/article/5e79b1d8bf0d4f5a884762ce3f6ab06c2018-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-19622-5https://doaj.org/toc/2045-2322Abstract Steviol glycosides from the leaves of the plant Stevia rebaudiana are high-potency natural sweeteners but suffer from a lingering bitterness. The Lactobacillus reuteri 180 wild-type glucansucrase Gtf180-ΔN, and in particular its Q1140E-mutant, efficiently α-glucosylated rebaudioside A (RebA), using sucrose as donor substrate. Structural analysis of the products by MALDI-TOF mass spectrometry, methylation analysis and NMR spectroscopy showed that both enzymes exclusively glucosylate the Glc(β1→C-19 residue of RebA, with the initial formation of an (α1→6) linkage. Docking of RebA in the active site of the enzyme revealed that only the steviol C-19 β-D-glucosyl moiety is available for glucosylation. Response surface methodology was applied to optimize the Gtf180-ΔN-Q1140E-catalyzed α-glucosylation of RebA, resulting in a highly productive process with a RebA conversion of 95% and a production of 115 g/L α-glucosylated products within 3 h. Development of a fed-batch reaction allowed further suppression of α-glucan synthesis which improved the product yield to 270 g/L. Sensory analysis by a trained panel revealed that glucosylated RebA products show a significant reduction in bitterness, resulting in a superior taste profile compared to RebA. The Gtf180-ΔN-Q1140E glucansucrase mutant enzyme thus is an efficient biocatalyst for generating α-glucosylated RebA variants with improved edulcorant/organoleptic properties.Evelien M. te PoeleTim DevlamynckManuel JägerGerrit J. GerwigDavy Van de WalleKoen DewettinckAnna K. H. HirschJohannis P. KamerlingWim SoetaertLubbert DijkhuizenNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-12 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Evelien M. te Poele Tim Devlamynck Manuel Jäger Gerrit J. Gerwig Davy Van de Walle Koen Dewettinck Anna K. H. Hirsch Johannis P. Kamerling Wim Soetaert Lubbert Dijkhuizen Glucansucrase (mutant) enzymes from Lactobacillus reuteri 180 efficiently transglucosylate Stevia component rebaudioside A, resulting in a superior taste
description	Abstract Steviol glycosides from the leaves of the plant Stevia rebaudiana are high-potency natural sweeteners but suffer from a lingering bitterness. The Lactobacillus reuteri 180 wild-type glucansucrase Gtf180-ΔN, and in particular its Q1140E-mutant, efficiently α-glucosylated rebaudioside A (RebA), using sucrose as donor substrate. Structural analysis of the products by MALDI-TOF mass spectrometry, methylation analysis and NMR spectroscopy showed that both enzymes exclusively glucosylate the Glc(β1→C-19 residue of RebA, with the initial formation of an (α1→6) linkage. Docking of RebA in the active site of the enzyme revealed that only the steviol C-19 β-D-glucosyl moiety is available for glucosylation. Response surface methodology was applied to optimize the Gtf180-ΔN-Q1140E-catalyzed α-glucosylation of RebA, resulting in a highly productive process with a RebA conversion of 95% and a production of 115 g/L α-glucosylated products within 3 h. Development of a fed-batch reaction allowed further suppression of α-glucan synthesis which improved the product yield to 270 g/L. Sensory analysis by a trained panel revealed that glucosylated RebA products show a significant reduction in bitterness, resulting in a superior taste profile compared to RebA. The Gtf180-ΔN-Q1140E glucansucrase mutant enzyme thus is an efficient biocatalyst for generating α-glucosylated RebA variants with improved edulcorant/organoleptic properties.
format	article
author	Evelien M. te Poele Tim Devlamynck Manuel Jäger Gerrit J. Gerwig Davy Van de Walle Koen Dewettinck Anna K. H. Hirsch Johannis P. Kamerling Wim Soetaert Lubbert Dijkhuizen
author_facet	Evelien M. te Poele Tim Devlamynck Manuel Jäger Gerrit J. Gerwig Davy Van de Walle Koen Dewettinck Anna K. H. Hirsch Johannis P. Kamerling Wim Soetaert Lubbert Dijkhuizen
author_sort	Evelien M. te Poele
title	Glucansucrase (mutant) enzymes from Lactobacillus reuteri 180 efficiently transglucosylate Stevia component rebaudioside A, resulting in a superior taste
title_short	Glucansucrase (mutant) enzymes from Lactobacillus reuteri 180 efficiently transglucosylate Stevia component rebaudioside A, resulting in a superior taste
title_full	Glucansucrase (mutant) enzymes from Lactobacillus reuteri 180 efficiently transglucosylate Stevia component rebaudioside A, resulting in a superior taste
title_fullStr	Glucansucrase (mutant) enzymes from Lactobacillus reuteri 180 efficiently transglucosylate Stevia component rebaudioside A, resulting in a superior taste
title_full_unstemmed	Glucansucrase (mutant) enzymes from Lactobacillus reuteri 180 efficiently transglucosylate Stevia component rebaudioside A, resulting in a superior taste
title_sort	glucansucrase (mutant) enzymes from lactobacillus reuteri 180 efficiently transglucosylate stevia component rebaudioside a, resulting in a superior taste
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/5e79b1d8bf0d4f5a884762ce3f6ab06c
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Glucansucrase (mutant) enzymes from Lactobacillus reuteri 180 efficiently transglucosylate Stevia component rebaudioside A, resulting in a superior taste

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