Sucrose phosphorylase (gene sucP) from Bifidobacterium adolescentis

Enzyme Description

Species

Extremophile

EC Number

2.4.1.7 ↗

Sequence

Length: 504 amino acids

MKNKVQLITYADRLGDGTIKSMTDILRTRFDGVYDGVHILPFFTPFDGADAGFDPIDHTKVDERLGSWDDVAELSKTHNIMVDAIVNHMSWESKQFQDVLAKGEESEYYPMFLTMSSVFPNGATEEDLAGIYRPRPGLPFTHYKFAGKTRLVWVSFTPQQVDIDTDSDKGWEYLMSIFDQMAASHVSYIRLDAVGYGAKEAGTSCFMTPKTFKLISRLREEGVKRGLEILIEVHSYYKKQVEIASKVDRVYDFALPPLLLHALSTGHVEPVAHWTDIRPNNAVTVLDTHDGIGVIDIGSDQLDRSLKGLVPDEDVDNLVNTIHANTHGESQAATGAAASNLDLYQVNSTYYSALGCNDQHYIAARAVQFFLPGVPQVYYVGALAGKNDMELLRKTNNGRDINRHYYSTAEIDENLKRPVVKALNALAKFRNELDAFDGTFSYTTDDDTSISFTWRGETSQATLTFEPKRGLGVDNTTPVAMLEWEDSAGDHRSDDLIANPPVVA

An Cerdobbel et al. (2011) — Increasing the thermostability of sucrose phosphorylase by a combination of sequence- and structure-based mutagenesis

Protein Engineering, Design and Selection · doi:10.1093/protein/gzr042 ↗ · Activity - Classical Stability - C50

64 measurements ▼

Database ID

UniProt: A0ZZH6 ↗

Sequence Annotation

Explicit - Provided GenBank Accession Number

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (64 measurements)

64 measurements — page 4 of 4

Mutation	Property	Assay	Solvent	Solvent Volume	Aqueous Reference	WT Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Q331E/R393N/D445P/D446T/Q460E/E485H	Stability - C50	Volume fraction of organic solvent for which 50% of activity, measured by absorbance spectrophotometry (reaction product α-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)	—	—	32	41	% v/v	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose , α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in % v/v)
R393N/D445P/D446T/Q460E/E485H	Stability - C50	Volume fraction of organic solvent for which 50% of activity, measured by absorbance spectrophotometry (reaction product α-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)	—	—	32	39	% v/v	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose , α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in % v/v)
R393N/Q460E/E485H	Stability - C50	Volume fraction of organic solvent for which 50% of activity, measured by absorbance spectrophotometry (reaction product α-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)	—	—	32	34	% v/v	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose , α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in % v/v)
Q460E/E485H	Stability - C50	Volume fraction of organic solvent for which 50% of activity, measured by absorbance spectrophotometry (reaction product α-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)	—	—	32	37	% v/v	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose , α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in % v/v)

WT	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	57	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	42	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	30	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	17	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	6	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	0	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	105	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
WT	Stability - C50	Volume fraction of organic solvent for which 50% of activity, measured by absorbance spectrophotometry (reaction product α-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)	—	—	—	32	% v/v	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in % v/v)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	54	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	60°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	0	5	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	64	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	60°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	67	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	60°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	73	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	60°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	81	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	60°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	88	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	60°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	105	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	60°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	110	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	60°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	113	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	60°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	113	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	60°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	0	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	65°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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)	33%	100	—	39	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	60°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	6	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	60°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	1	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	60°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	0	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	60°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	0	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	60°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	103	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	65°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	90	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	65°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	80	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	65°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	41	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	65°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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)	27.5%	100	—	21	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	65°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	13	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	65°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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)	32.5%	100	—	7	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	65°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	—	3	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	65°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	57	94	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	105	115	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	57	65	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	42	57	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	30	36	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	17	21	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	6	3	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	0	0	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
R393N/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	105	108	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
R393N/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	57	67	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
R393N/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	42	49	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
R393N/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	30	42	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
R393N/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	17	15	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
R393N/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	6	5	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
R393N/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	0	0	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	105	133	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	6	11	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	42	63	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	30	39	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	17	26	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	6	5	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	0	1	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	105	135	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	57	95	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	42	72	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	30	52	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Activity - Classical	Activity measured by absorbance spectrophotometry (reaction product α-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%	100	17	33	%	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in %)
Q331E/R393N/D445P/D446T/Q460E/E485H	Stability - C50	Volume fraction of organic solvent for which 50% of activity, measured by absorbance spectrophotometry (reaction product α-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)	—	—	32	41	% v/v	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in % v/v)
R393N/D445P/D446T/Q460E/E485H	Stability - C50	Volume fraction of organic solvent for which 50% of activity, measured by absorbance spectrophotometry (reaction product α-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)	—	—	32	39	% v/v	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in % v/v)
R393N/Q460E/E485H	Stability - C50	Volume fraction of organic solvent for which 50% of activity, measured by absorbance spectrophotometry (reaction product α-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)	—	—	32	34	% v/v	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in % v/v)
Q460E/E485H	Stability - C50	Volume fraction of organic solvent for which 50% of activity, measured by absorbance spectrophotometry (reaction product α-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)	—	—	32	37	% v/v	50 mM phosphate buffer, 2 mM EDTA, 10 mM MgSO4, 2 mM β-NAD, 10 mM glucose-1.6-diphosphate, 1.2 U PGM, 1.2 U G6P-DH	7	37°C	50 mM Phosphate, 50 mM Sucrose	fructose, α-D-glucose-1-phosphate (G1P)	—	—	Classical aqueous control (in % v/v)

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Mutations in this dataset (5)

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

Visualization : Activity — Classical

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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 — C₅₀

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

Structure

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