Sucrose Phosphorylase and Related Enzymes in Glycoside Hydrolase Family 13: Discovery, Application and Engineering

Abstract

Sucrose phosphorylases are carbohydrate-active enzymes with outstanding potential for the biocatalytic conversion of common table sugar into products with attractive properties. They belong to the glycoside hydrolase family GH13, where they are found in subfamily 18. In bacteria, these enzymes catalyse the phosphorolysis of sucrose to yield α-glucose 1-phosphate and fructose. However, sucrose phosphorylases can also be applied as versatile transglucosylases for the synthesis of valuable glycosides and sugars because their broad promiscuity allows them to transfer the glucosyl group of sucrose to a diverse collection of compounds other than phosphate. Numerous process and enzyme engineering studies have expanded the range of possible applications of sucrose phosphorylases ever further. Moreover, it has recently been discovered that family GH13 also contains a few novel phosphorylases that are specialised in the phosphorolysis of sucrose 6F-phosphate, glucosylglycerol or glucosylglycerate. In this review, we provide an overview of the progress that has been made in our understanding and exploitation of sucrose phosphorylases and related enzymes over the past ten years.

On the donor substrate dependence of group‐transfer reactions by hydrolytic enzymes: Insight from kinetic analysis of sucrose phosphorylase‐catalyzed transglycosylation

“…Both enzymes have been well characterized previously (Franceus & Desmet, 2020;Goedl, Sawangwan, Wildberger, & Nidetzky, 2010;, and the LmSucP is used for commercial production of GG . Guided by kinetic framework analysis developed here, we show that in enzymatic reactions with G1P, hydrolysis takes place not only from the covalent glucosyl-enzyme intermediate (E-Glc), but additionally from a noncovalent complex of E-Glc and G1P which unlike E-Glc cannot be intercepted by glycerol.…”

Disaccharide phosphorylases: Structure, catalytic mechanisms and directed evolution

“…The structural microenvironment of subsite +1 can undergo drastic rearrangements during the catalytic process because of its unique double displacement mechanism. This conformational flexibility is probably responsible for the activity promiscuity of SPs [ 58 ].…”

“…The structure-based rational design was then developed to introduce additional salt bridges and alleviate a potential electrostatic repulsion at the dimer interface. Combining all mutations, the half-life time of BaSP at 60 °C was increased dramatically from 24 h to 62 h [ 58 ]. Moreover, for CtCBP, site-directed mutagenesis based on structure-guided homology analysis and random mutagenesis was applied to improve thermal stability and temperature optimum of the enzyme.…”

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

“…We also considered the substantial replication potential of a continuous process technology for biocatalytic glycosylation from sucrose. A number of α-D-glucoside products are accessible via SucP-catalyzed glycosylation (Goedl et al 2010;Franceus and Desmet 2020), several of which (e.g., kojibiose (Beerens et al 2017)) have considerable significance for industrial production. Having in mind material properties of the cell-polymer composite important for continuous bioprocessing (e.g., swelling, hardness, elasticity), we examined two principal routes of cell encapsulation.…”