Enzymatic Synthesis of Galacto-oligosaccharides in an Organic-Aqueous Biphasic System by a Novel β-Galactosidase from a Metagenomic Library

Prebiotic galacto-oligosaccharides (GOS) were effectively synthesized from lactose in organic-aqueous biphasic media by,a novel metagenome-derived beta-galactosidase BgaP412. A maximum GOS of 46.6% (w/w) was achieved with 75.4% lactose conversion rate in the cyclohexane/buffer system [95:5 (v/v) cyclohexane/buffer] under the optimum reaction conditions (initial lactose concentration = 30% (w/v), T = 50 degrees C, pH 7.0, and t = 8 h). The corresponding productivity of GOS was approximately 17.5 g L-1 h(-1). The GOS mixture consisted of tri-, tetra-, and pentasaccharides. Trisaccharides were the chief component of reaction products. These experimental results showed that a low water content, a high initial lactose concentration, and an elevated reaction temperature could significantly promote the transgalactosylation activity of beta-galactosidase BgaP412; at the same time, the enhanced GOS yield in an organic-aqueous biphasic system is because of the fact that thermodynamic equilibrium can be shifted to the synthetic direction by reversing the normal hydrolysis.