BRENDA - Enzyme Database

Redesign of the active site of sucrose phosphorylase through a clash-induced cascade of loop shifts

Kraus, M.; Grimm, C.; Seibel, J.; ChemBioChem 17, 33-36 (2016)

Data extracted from this reference:

Crystallization (Commentary)
EC Number
Crystallization
Organism
2.4.1.7
recombinant enzyme mutant Q345F, in the presence of sucrose, and in complex with glucose, X-ray diffraction structure determination and analysis at 2.7 A resolution, PDB ID 5C8B
Bifidobacterium adolescentis
Engineering
EC Number
Amino acid exchange
Commentary
Organism
2.4.1.7
additional information
redesign of the active site of sucrose phosphorylase through a clash-induced cascade of loop shifts
Bifidobacterium adolescentis
2.4.1.7
Q345F
site-directed mutagenesis, the mutation allows efficient glucosylation of resveratrol, (+)-catechin and (-)-epicatechin in yields of up to 97% whereas the wild-type enzyme favours sucrose hydrolysis. The crystal structure of the variant reveals a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors. The generation of this channel can be explained in terms of a cascade of structural changes arising from the Q345F exchange, structural changes in the active site of BaSP Q345F, modeling, overview
Bifidobacterium adolescentis
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.4.1.7
sucrose + phosphate
Bifidobacterium adolescentis
-
D-fructose + alpha-D-glucose 1-phosphate
-
-
r
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
2.4.1.7
Bifidobacterium adolescentis
Q84HQ2
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.4.1.7
additional information
the wild-type enzyme shows poor activity with flavonoids or stilbenoids, e.g. resveratrol, (+)-catechin and (-)-epicatechin, while the compounds are substrates of the enzyme mutant Q345F
735952
Bifidobacterium adolescentis
?
-
-
-
-
2.4.1.7
sucrose + (+)-catechin
activity of enzyme mutant Q345F
735952
Bifidobacterium adolescentis
D-fructose + (+)-catechin 3'-O-alpha-D-glucoside + (+)-catechin 3',5-O-alpha-D-diglucoside
-
-
-
?
2.4.1.7
sucrose + (-)-epicatechin
activity of enzyme mutant Q345F
735952
Bifidobacterium adolescentis
D-fructose + (-)-epicatechin 3'-O-alpha-D-glucoside + (-)-epicatechin 5-O-alpha-D-glucoside + (-)-epicatechin 3',5-O-alpha-D-diglucoside
-
-
-
?
2.4.1.7
sucrose + phosphate
-
735952
Bifidobacterium adolescentis
D-fructose + alpha-D-glucose 1-phosphate
-
-
-
r
2.4.1.7
sucrose + phosphate
activity of wild-type enzyme and mutant Q345F
735952
Bifidobacterium adolescentis
D-fructose + alpha-D-glucose 1-phosphate
-
-
-
r
2.4.1.7
sucrose + resveratrol
activity of enzyme mutant Q345F
735952
Bifidobacterium adolescentis
D-fructose + resveratrol 3-O-alpha-D-glucoside
-
-
-
?
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
2.4.1.7
recombinant enzyme mutant Q345F, in the presence of sucrose, and in complex with glucose, X-ray diffraction structure determination and analysis at 2.7 A resolution, PDB ID 5C8B
Bifidobacterium adolescentis
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
2.4.1.7
additional information
redesign of the active site of sucrose phosphorylase through a clash-induced cascade of loop shifts
Bifidobacterium adolescentis
2.4.1.7
Q345F
site-directed mutagenesis, the mutation allows efficient glucosylation of resveratrol, (+)-catechin and (-)-epicatechin in yields of up to 97% whereas the wild-type enzyme favours sucrose hydrolysis. The crystal structure of the variant reveals a widened access channel with a hydrophobic aromatic surface that is likely to contribute to the improved activity towards aromatic acceptors. The generation of this channel can be explained in terms of a cascade of structural changes arising from the Q345F exchange, structural changes in the active site of BaSP Q345F, modeling, overview
Bifidobacterium adolescentis
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
2.4.1.7
sucrose + phosphate
Bifidobacterium adolescentis
-
D-fructose + alpha-D-glucose 1-phosphate
-
-
r
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
2.4.1.7
additional information
the wild-type enzyme shows poor activity with flavonoids or stilbenoids, e.g. resveratrol, (+)-catechin and (-)-epicatechin, while the compounds are substrates of the enzyme mutant Q345F
735952
Bifidobacterium adolescentis
?
-
-
-
-
2.4.1.7
sucrose + (+)-catechin
activity of enzyme mutant Q345F
735952
Bifidobacterium adolescentis
D-fructose + (+)-catechin 3'-O-alpha-D-glucoside + (+)-catechin 3',5-O-alpha-D-diglucoside
-
-
-
?
2.4.1.7
sucrose + (-)-epicatechin
activity of enzyme mutant Q345F
735952
Bifidobacterium adolescentis
D-fructose + (-)-epicatechin 3'-O-alpha-D-glucoside + (-)-epicatechin 5-O-alpha-D-glucoside + (-)-epicatechin 3',5-O-alpha-D-diglucoside
-
-
-
?
2.4.1.7
sucrose + phosphate
-
735952
Bifidobacterium adolescentis
D-fructose + alpha-D-glucose 1-phosphate
-
-
-
r
2.4.1.7
sucrose + phosphate
activity of wild-type enzyme and mutant Q345F
735952
Bifidobacterium adolescentis
D-fructose + alpha-D-glucose 1-phosphate
-
-
-
r
2.4.1.7
sucrose + resveratrol
activity of enzyme mutant Q345F
735952
Bifidobacterium adolescentis
D-fructose + resveratrol 3-O-alpha-D-glucoside
-
-
-
?