Synthesis of Galactosyl Mannitol Derivative Using β-Galactosidase from Kluyveromyces lactis

Abstract

The purpose of the study was to identify the infl uence of reactive mixture concentration (23-48 g/100 mL), pH (6.5-9.0), presence of NaCl (0.05-0.25 mol/L) and enzyme dose (2850-28,500 LAU/100 g of lactose) on the synthesis of galactosyl mannitol derivative using β-galactosidase from Kluyveromyces lactis. The use of the enzyme dose ranging from 2850 to 11,400 LAU/100 g lactose allowed obtaining gal-mannitol at the level of 21.8% total saccharides; higher doses intensifi ed product decomposition. An increase in the concentration of the reactive mixture had a positive impact on the quantity of gal-mannitol obtained every single time, i.e. 4.39 g were obtained from 100 mL of a 23 g/100 mL solution and over 10 g were obtained from a 48 g/100 mL solution. A relatively low increase in product quantity (by ca. 5%) occurred after the pH was increased from 6.5 to 9.0. The use of NaCl rendered better results. An increase in the maximum content of gal-mannitol in the total sugar by 12.8% was observed at the concentration of 0.25 mol/L.

Active Site Architecture and Reaction Mechanism Determination of Cold Adapted β-d-galactosidase from Arthrobacter sp. 32cB

“…Similar to Ecol βDG, it consists of 1032 aa and its functional form is a homotetramer. Not only it catalyzes lactose hydrolysis, but also transglycosylation reaction that results in formation of galactose derivatives, among other alkyl-galactosides [11], gal-mannitol, [12], and bionic acids [13].…”

Application of Transgalactosylation Activity of β-Galactosidase from Kluyveromyces lactis for the Synthesis of Ascorbic Acid Galactoside

“…Other molecules possessing a hydroxyl group can be used as acceptor (e.g. sugars such as fructose [ 33 ], polyols such as glycerol [ 34 ], mannitol [ 35 ] and polyhydroxy acids such as gluconic acid [ 36 ]). In this case, the galactosyl moiety from lactose was transferred onto an ascorbic acid molecule as a result of activity of β-galactosidase produced by K. lactis .…”

“…However, in the case of ascorbic acid galactoside, even a significant change in the enzyme dose did not contributed to visible differences in the course of reaction. Generally, the increase of enzyme dose leads to shortening the time of transgalactosylation reaction and simultaneously further-reaching hydrolysis of transgalactosylation products [ 35 , 42 ]. Considering formation of GOS, Martínez-Villaluenga et al [ 42 ] achieved the highest trisaccharide content with the lowest tested amount of Lactozym 3000L HP G (β-galactosidase from K. lactis ).…”

“…However, the value diminished after that time (due to the hydrolysis mentioned above). Studies on synthesis of galactosyl mannitol derivative (with the same enzyme preparation) performed by Klewicki et al [ 35 ] demonstrated that raising the enzyme dose from an optimum of 11,400 LAU/100 g lactose to 22,800 caused almost a 7% drop in the highest concentration of product. In the presented research, this effect is not significant.…”

Hydrolysis of Lactose and Transglycosylation of Selected Sugar Alcohols by LacA β-Galactosidase from Lactobacillus plantarum WCFS1

“…In this context, it has been demonstrated that not only saccharides may act as the acceptors of galactosyl moieties. In fact, polyols (also known as sugar alcohols) could be, under specific conditions, better acceptors than carbohydrates favoring transglycosylation reaction over hydrolysis 10 , producing galactosyl derivatives of polyols (Gal-polyols) 11 .…”