Solvents derived from glycerol modify classical regioselectivity in the enzymatic synthesis of disaccharides with Biolacta β-galactosidase

Abstract

Green solvents made from glycerol change the classical regioselectivity of Biolacta N o 5 -galactosidase, from (1→4) toward (1→6) linkages when 2M concentration was used. In order to explain these results, the non-proteic compounds present in Biolacta preparation were separated by precipitation with 10 ammonium sulfate and the remaining protein extract was used to set reactions with appropriate organic solvents to find that the regioselectivity towards the β(1 6) isomer is retained. According to proteomic analysis, a 98% homology between Streptococcus pneumoniae and Biolacta -galactosidase preparation was found. With these data, molecular modelling was done and predicts a tridimensional interaction in the enzyme active site with the donor (GlcNAc) and the water-solvent mixture that explains this 15 phenomenon.

Synthesis of 3-alkoxypropan-1,2-diols from glycidol: experimental and theoretical studies for the optimization of the synthesis of glycerol derived solvents

“…Thus, fluorinated glycerol derivatives, such as the closely related 1,3-bis(2,2,2trifluoroethoxy)propan-2-ol [3F.0.3F], have been successfully used as reaction media in epoxidation and enzymatic reactions. 38,[47][48][49]…”

Highly efficient and regioselective enzymatic synthesis of β-(1→3) galactosides in biosolvents

“…Recently, it has been shown that the use of green solvents, 20,21 such as biomass derived solvents or ionic liquids, as co-solvents in the enzymatic reaction lead to a variation of the enzyme activity. [22][23][24][25][26][27][28][29] Biomass derived solvents show some new and attractive advantages, as there are made from a renewable source, they are (presumably) less toxic, exhibit low volatility, and possess tuneable physicochemical properties. 20,21,[30][31][32] We have previously described the use of some of these biomass derived solvents in the transglycosylation reaction catalyzed by Biolacta b-galactosidase, using p-nitrophenyl-b-D-galactopyranoside (p-NP-b-Gal) as donor and GlcNAc as acceptor, changing the regioselectivity of the enzyme from a mixture of b(1A4) and b(1A6) towards the synthesis of only b(1A6) oligosaccharides.…”

Enzymatic synthesis of novel fructosylated compounds by Ffase from Schwanniomyces occidentalis in green solvents

“…Reactions (500 mL) contained 10 U mL À1 -standard b-fructofuranosidase activity, 200 g L À1 sucrose in 0.2 M sodium acetate buffer pH 5.5, and 30% (v/v) nal concentration of solvents, except for GCL and DMA at 2 M. 28,55,56 Spain). Data obtained aer 65 min total analysis time were processed by Borwin soware (v. 1.5; Jasco Inc.).…”