High-Yield Biosynthesis of Glucosylglycerol through Coupling Phosphorolysis and Transglycosylation Reactions

Abstract

Glucosylglycerol is a powerful osmolyte that has attracted attention as a useful moisturizing ingredient in the cosmetic industry. This study demonstrates two artificially designed synthetic routes for manufacturing glucosylglycerol by combining phosphorolysis and transglycosylation reactions. The overall Gibbs energy change of the synthetic routes was negative, indicating that they are thermodynamically favorable. In vitro biosystems were constructed through combining the phosphorolysis ability of sucrose/maltose phosphorylase and the transglycosylation capacity of glucosylglycerol phosphorylases from different organisms. A near-stoichiometric conversion of sucrose and glycerol with a high product yield of 98% was achieved under optimal reaction conditions. The large-scale glucosylglycerol production of this biosystem was investigated under a high concentration of substrates (2 mol/L sucrose and 2.4 mol/L glycerol), and the titer reached 1.78 mol/L (452 g/L) with a productivity of 24.3 g/L/h. To the best of our knowledge, this value presented the highest glucosylglycerol production level until now, which indicated a great industrial application potential for glucosylglycerol manufacturing.

Continuous process technology for glucoside production from sucrose using a whole cell-derived solid catalyst of sucrose phosphorylase

“…Operation of the packed-bed reactor was also possible at high substrate loadings (up to 300 g L −1 sucrose), delivering excellent process productivities (Table 1) (Straathof 2014;De Santis et al 2020). The calculated STY was higher than the highest glucosyl-glycerol productivity (24.3 g L −1 h −1 ) reported by now, which was achieved with a coupled phosphorolysis and trans-glycosylation two-enzyme system (Zhang et al 2020). The achieved glucosyl-glycerol titer in the Zhang et al's study was higher (452 g L −1 ) than that reached here.…”

“…The achieved glucosyl-glycerol titer in the Zhang et al's study was higher (452 g L −1 ) than that reached here. However, direct comparison of our study with that of Zhang et al (2020) is difficult, since different enzyme systems, biocatalyst formulations, and reaction set-ups were applied. The productivities achieved with the packed-bed reactor were also higher than for comparable processes using immobilized cells.…”

“…A number of enzymes have been immobilized via encapsulation of whole cells of the native microorganism. Among them was sucrose phosphorylase (SucP) (Vandamme et al 1987 ) which is also the target of the current inquiry, performed in the actual context of industrial biomanufacturing of the cosmetic ingredient 2-α- d -glucosyl-glycerol (2-GG) (Goedl et al 2008 ; Luley-Goedl et al 2010 ; Tan et al 2016 ; Roenneke et al 2018 ; Zhang et al 2020 ).…”

Engineering of a Thermostable Biocatalyst for the Synthesis of 2‐O‐Glucosylglycerol

“…Very recently, an alternative two‐enzyme process was suggested where the native phosphorolytic reaction of LmSP was coupled to the activity of a natural glucosylglycerol phosphorylase (GGoP), which can generate the osmolyte very efficiently but requires α‐glucose 1‐phosphate as donor substrate instead of sucrose [11] . This coupled system was claimed to reach far higher productivities than the current industrial process even at low glycerol concentrations, but the low thermostability of LmSP was identified as an important hurdle of this reaction set‐up as well [12] . Hence, there is a clear and economically relevant need for an SP that is better suited for the industrial production of 2‐ O ‐glucosylglycerol.…”

“… [11] This coupled system was claimed to reach far higher productivities than the current industrial process even at low glycerol concentrations, but the low thermostability of LmSP was identified as an important hurdle of this reaction set‐up as well. [12] Hence, there is a clear and economically relevant need for an SP that is better suited for the industrial production of 2‐ O ‐glucosylglycerol.…”