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

Autor: 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., Sanders P; Eurofins Expertise Centre for Complex Carbohydrates and Chemistry, PO Box 766, 8440 AT, Heerenveen, The Netherlands., 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., 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.
Jazyk: angličtina
Zdroj: Applied microbiology and biotechnology [Appl Microbiol Biotechnol] 2024 May 29; Vol. 108 (1), pp. 349. Date of Electronic Publication: 2024 May 29.
DOI: 10.1007/s00253-024-13183-6
Abstrakt: 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.
(© 2024. The Author(s).)
Databáze: MEDLINE
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