Kinetics and products of Thermotoga maritima β-glucosidase with lactose and cellobiose.


Journal

Applied microbiology and biotechnology
ISSN: 1432-0614
Titre abrégé: Appl Microbiol Biotechnol
Pays: Germany
ID NLM: 8406612

Informations de publication

Date de publication:
29 May 2024
Historique:
received: 08 01 2024
accepted: 14 05 2024
revised: 09 05 2024
medline: 29 5 2024
pubmed: 29 5 2024
entrez: 29 5 2024
Statut: epublish

Résumé

Galacto-oligosaccharides (GOS) are prebiotic compounds that are mainly used in infant formula to mimic bifidogenic effects of mother's milk. They are synthesized by β-galactosidase enzymes in a trans-glycosylation reaction with lactose. Many β-galactosidase enzymes from different sources have been studied, resulting in varying GOS product compositions and yields. The in vivo role of these enzymes is in lactose hydrolysis. Therefore, the best GOS yields were achieved at high lactose concentrations up to 60%wt, which require a relatively high temperature to dissolve. Some thermostable β-glucosidase enzymes from thermophilic bacteria are also capable of using lactose or para nitrophenyl-galactose as a substrate. Here, we describe the use of the β-glucosidase BglA from Thermotoga maritima for synthesis of oligosaccharides derived from lactose and cellobiose and their detailed structural characterization. Also, the BglA enzyme kinetics and yields were determined, showing highest productivity at higher lactose and cellobiose concentrations. The BglA trans-glycosylation/hydrolysis ratio was higher with 57%wt lactose than with a nearly saturated cellobiose (20%wt) solution. The yield of GOS was very high, reaching 72.1%wt GOS from lactose. Structural elucidation of the products showed mainly β(1 → 3) and β(1 → 6) elongating activity, but also some β(1 → 4) elongation was observed. The β-glucosidase BglA from T. maritima was shown to be a very versatile enzyme, producing high yields of oligosaccharides, particularly GOS from lactose. KEY POINTS: • β-Glucosidase of Thermotoga maritima synthesizes GOS from lactose at very high yield. • Thermotoga maritima β-glucosidase has high activity and high thermostability. • Thermotoga maritima β-glucosidase GOS contains mainly (β1-3) and (β1-6) linkages.

Identifiants

pubmed: 38809317
doi: 10.1007/s00253-024-13183-6
pii: 10.1007/s00253-024-13183-6
doi:

Substances chimiques

Lactose J2B2A4N98G
Cellobiose 16462-44-5
beta-Glucosidase EC 3.2.1.21
Oligosaccharides 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

349

Informations de copyright

© 2024. The Author(s).

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Auteurs

Geert A Ten Kate (GA)

Microbial Physiology, Groningen Biomolecular Sciences and Biotechnology Institute (GBB), University of Groningen, Nijenborgh 7, 9747 AG, Groningen, The Netherlands.
Royal FrieslandCampina, Stationsplein 4, 3818 LE, Amersfoort, The Netherlands.

Peter Sanders (P)

Eurofins Expertise Centre for Complex Carbohydrates and Chemistry, PO Box 766, 8440 AT, Heerenveen, The Netherlands.

Lubbert Dijkhuizen (L)

Microbial Physiology, Groningen Biomolecular Sciences and Biotechnology Institute (GBB), University of Groningen, Nijenborgh 7, 9747 AG, Groningen, The Netherlands.
CarbExplore Research BV, Zernikelaan 8, 9747 AA, Groningen, The Netherlands.

Sander S van Leeuwen (SS)

Microbial Physiology, Groningen Biomolecular Sciences and Biotechnology Institute (GBB), University of Groningen, Nijenborgh 7, 9747 AG, Groningen, The Netherlands. s.s.van.leeuwen@rug.nl.
Department of Laboratory Medicine, University of Groningen, University Medical Center Groningen, Hanzeplein 1, EA30, 9713 GZ, Groningen, The Netherlands. s.s.van.leeuwen@rug.nl.
Van Hall Larenstein, University of Applied Sciences, Agora 1, P.O. box 1528, 8901 BV, Leeuwarden, The Netherlands. s.s.van.leeuwen@rug.nl.

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