Sucrose phosphorylase (gene sucP) from Bifidobacterium adolescentis

Enzyme Description

Species
Bifidobacterium adolescentis β†—
Extremophile
No
EC Number
2.4.1.7 β†—

Sequence

Length: 504 amino acids
MKNKVQLITYADRLGDGTIKSMTDILRTRFDGVYDGVHILPFFTPFDGADAGFDPIDHTKVDERLGSWDDVAELSKTHNIMVDAIVNHMSWESKQFQDVLAKGEESEYYPMFLTMSSVFPNGATEEDLAGIYRPRPGLPFTHYKFAGKTRLVWVSFTPQQVDIDTDSDKGWEYLMSIFDQMAASHVSYIRLDAVGYGAKEAGTSCFMTPKTFKLISRLREEGVKRGLEILIEVHSYYKKQVEIASKVDRVYDFALPPLLLHALSTGHVEPVAHWTDIRPNNAVTVLDTHDGIGVIDIGSDQLDRSLKGLVPDEDVDNLVNTIHANTHGESQAATGAAASNLDLYQVNSTYYSALGCNDQHYIAARAVQFFLPGVPQVYYVGALAGKNDMELLRKTNNGRDINRHYYSTAEIDENLKRPVVKALNALAKFRNELDAFDGTFSYTTDDDTSISFTWRGETSQATLTFEPKRGLGVDNTTPVAMLEWEDSAGDHRSDDLIANPPVVA
An Cerdobbel et al. (2011) β€” Increasing the thermostability of sucrose phosphorylase by a combination of sequence- and structure-based mutagenesis
Protein Engineering, Design and Selection  Β· doi:10.1093/protein/gzr042 β†—  Β· Activity - Classical Stability - C50
64 measurements ▼
Database ID
UniProt: A0ZZH6 β†—
Sequence Annotation
Explicit - Provided GenBank Accession Number
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (64 measurements)

64 measurements β€” page 1 of 4
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Extraction method Data type Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
WT Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 30 % v/v 100 β€” 57 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 37Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
WT Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 35 % v/v 100 β€” 42 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 37Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
WT Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 40 % v/v 100 β€” 30 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 37Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
WT Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 45 % v/v 100 β€” 17 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 37Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
WT Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 50 % v/v 100 β€” 6 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 37Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
WT Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 55 % v/v 100 β€” 0 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 37Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
WT Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 20 % v/v 100 β€” 105 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 37Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
WT Stability - C50 Volume fraction of organic solvent for which 50% of activity, measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) Dimethyl Sulfoxide (DMSO) β€” β€” β€” 32 % v/v Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 37Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in % v/v)
Q331E/R393N/D445P/D446T/Q460E/E485H Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 32 % v/v 100 β€” 54 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 60Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 55 % v/v 100 0 5 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 37Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 31 % v/v 100 β€” 64 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 60Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 30 % v/v 100 β€” 67 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 60Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 29 % v/v 100 β€” 73 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 60Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 28 % v/v 100 β€” 81 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 60Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 27 % v/v 100 β€” 88 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 60Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 25 % v/v 100 β€” 105 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 60Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 20 % v/v 100 β€” 110 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 60Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 15 % v/v 100 β€” 113 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 60Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 10 % v/v 100 β€” 113 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 60Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H Activity - Classical Activity measured by absorbance spectrophotometry (reaction product Ξ±-D-glucose-1-phosphate transformed into glucose-6-phosphate by Phosphoglucomutase dehydrogenation catalyzed by Glucose-6-phosphate dehydrogenase absorbance measurement, 340 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 40 % v/v 100 β€” 0 % Digital Continuous 50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM Ξ²-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH 7 65Β°C 50 mM Phosphate, 50 mM Sucrose Fructose , Ξ±-D-glucose-1-phosphate (G1P) β€” β€” Classical aqueous control (in %)
β€Ή Prev
1 2 3 4
Next β€Ί

Mutations in this dataset (5)

WT Q331E/R393N/D445P/D446T/Q460E/E485H R393N/D445P/D446T/Q460E/E485H R393N/Q460E/E485H Q460E/E485H

Visualization : Activity β€” Classical

One bar per measurement. Colour = solvent, shade = solvent volume. β€” β€” β€” Reference value. Hover for details.

Mutation Effect

Mutation impact on enzyme stability and function in the presence of organic solvent: comparison of wild-type and mutant values in identical conditions.

Visualization : Stability β€” C50

One bar per measurement. Colour = solvent, shade = temperature. Hover for details.

Structure

Drag to rotate Β· Scroll to zoom Β· Right-click drag to pan Β· Powered by Mol*