Literature summary for 2.4.1.7 extracted from

Goedl, C.; Schwarz, A.; Mueller, M.; Brecker, L.; Nidetzky, B.
Mechanistic differences among retaining disaccharide phosphorylases: insights from kinetic analysis of active site mutants of sucrose phosphorylase and alpha,alpha-trehalose phosphorylase (2008), Carbohydr. Res., 343, 2032-2040.

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Protein Variants

Protein Variants	Comment	Organism
D196A	site-directed mutagenesis, the purified D196A mutant shows 40% reduced activity compared to the wild-type in phosphorolysis and synthesis of sucrose as well as arsenolysis of alpha-glucose 1-phosphate, however, with azide as an alternative nucleophile, the conversion of alpha-glucose 1-phosphate proceeds at a slow rate and results in the formation of product glucose 1-azide with a beta-anomeric configuration, activity enhancement in the D196A mutant results from the direct participation of azide in the now inverting, single displacement-like mechanism of glucosyl transfer, overview	Leuconostoc mesenteroides
D295N	site-directed mutagenesis, the mutant shows 0.01% of the wild-type sucrose phosphorolysis activity, meaning a reduction by 20000fold, but regaines activity by heat treatment for 10 min at 100°C, caused by a partial deamidation of D295. The catalytic defect resulting from the substitution of Asp295 is independent of the leaving group ability and nucleophilic reactivity of the substrate	Leuconostoc mesenteroides
E237Q	site-directed mutagenesis, the mutant shows altered pH-dependence compared to the wild-type enzyme	Leuconostoc mesenteroides

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
sucrose + phosphate	Leuconostoc mesenteroides	-	D-fructose + alpha-D-glucose 1-phosphate	-	?

Organism

Organism	UniProt	Comment	Textmining
Leuconostoc mesenteroides	-	-	-

Reaction

Reaction	Comment	Organism	Reaction ID
sucrose + phosphate = D-fructose + alpha-D-glucose 1-phosphate	reaction mechanism, overview, sucrose phosphorylase utilizes a glycoside hydrolase-like double displacement mechanism to convert its disaccharide substrate and phosphate into alpha-D-glucose 1-phosphate and D-fructose, roles of Asp196 and Glu237 as catalytic nucleophile and acid-base, respectively. The side chain of Asp295 facilitates the catalytic steps of glucosylation and deglucosylation of Asp196 through a strong hydrogen bond with the 2-hydroxyl of the glucosyl oxocarbenium ion-like species formed in the transition states flanking the beta-glucosyl enzyme intermediate	Leuconostoc mesenteroides	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
sucrose + phosphate	-	Leuconostoc mesenteroides	D-fructose + alpha-D-glucose 1-phosphate	-	?
sucrose + phosphate	differential binding of fructose and phosphate as leaving group/nucleophile of the reaction, structure, Asp295-transition state stabilization through hydrogen bonding, overview	Leuconostoc mesenteroides	D-fructose + alpha-D-glucose 1-phosphate	-	?

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
additional information	-	deglucosylation of mutant E237Q is pH-independent	Leuconostoc mesenteroides
5.6	7.2	wild-type enzyme, optimal pH for glycosylation	Leuconostoc mesenteroides

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Release 2023.1 (January 2023)