β-glycosidase (gene MeBglD2) from a soil metagenomic library

Enzyme Description

Species

Na (soil metagenomic library)

Extremophile

EC Number

3.2.1.21 ↗

Sequence

Length: 455 amino acids

MSTQNEPHRFPPDFQWGVATSSYQIEGAVEADGRSPSIWDTFCARPGAIADGSTGAIANDHYHRYREDIAIMKQLGVNAYRFSIAWPRILPDGRGRVNQAGVDFYERLVDSLLEQGIEPYATLYHWDMPQVQHDRTPWYDRGVVDAFVEYTDVITRRLSDRVKYWMTLNEPWVISFLGYGAGEHAPGLRDKELYLRAAHHVLLAHGKAMPVIRANGNAQTKAGIVLNLNWVNAASDSPEDQAAARRYDQFFNRWFAEPLYNGRYPEELLEWYGRDLVPVQPGDFDIITTPTDFLAVNYYARTTVKAGSTDPMLQVDFVRPPGEYTAMDWEVYPQGLYNILNWLHTDYAPPALYVTENGAAYDDQVSAAGEVDDPQRLAYLEGHFEAAYRAIQAGIPLKGYFVWSLMDNFEWGRGFEKRFGIVFVDYATQQRIIKRSGKWFSQVTRANGLPAPQTT

Tomohiko Matsuzawa et al. (2017) — Improved thermostability of a metagenomic glucose-tolerant β-glycosidase based on its X-ray crystal structure

Applied Microbiology and Biotechnology · doi:10.1007/s00253-017-8525-9 ↗ · Activity - Classical

12 measurements ▼

Database ID

UniProt: A0A1E1FFN6 ↗

Sequence Annotation

Explicit - Provided

Protein Source

Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (12 measurements)

12 measurements

Mutation	Property	Assay	Solvent	Solvent Volume	Aqueous Reference	WT Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	10%	100.0	—	20.4	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose , p-Nitrophenol	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100.0	—	74.9	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose , p-Nitrophenol	—	—	Classical aqueous control (in %)
H8L	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	10%	100.0	20.4	25.7	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose , p-Nitrophenol	—	—	Classical aqueous control (in %)
N59C	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	10%	100.0	20.4	119.0	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose , p-Nitrophenol	—	—	Classical aqueous control (in %)
A295G	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	10%	100.0	20.4	57.5	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose , p-Nitrophenol	—	—	Classical aqueous control (in %)
N59C/A295G	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	10%	100.0	20.4	127.0	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose , p-Nitrophenol	—	—	Classical aqueous control (in %)
H8L/N59C/A295G	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	10%	100.0	20.4	129.0	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose , p-Nitrophenol	—	—	Classical aqueous control (in %)
H8L	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100.0	74.9	75.3	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose , p-Nitrophenol	—	—	Classical aqueous control (in %)
N59C	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100.0	74.9	101.0	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose , p-Nitrophenol	—	—	Classical aqueous control (in %)
A295G	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100.0	74.9	82.8	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose , p-Nitrophenol	—	—	Classical aqueous control (in %)
N59C/A295G	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100.0	74.9	103.0	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose , p-Nitrophenol	—	—	Classical aqueous control (in %)
H8L/N59C/A295G	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100.0	74.9	104.0	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose , p-Nitrophenol	—	—	Classical aqueous control (in %)

WT	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	10%	100.0	—	20.4	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose, p-Nitrophenol	—	—	Classical aqueous control (in %)
WT	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100.0	—	74.9	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose, p-Nitrophenol	—	—	Classical aqueous control (in %)
H8L	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	10%	100.0	20.4	25.7	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose, p-Nitrophenol	—	—	Classical aqueous control (in %)
N59C	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	10%	100.0	20.4	119.0	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose, p-Nitrophenol	—	—	Classical aqueous control (in %)
A295G	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	10%	100.0	20.4	57.5	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose, p-Nitrophenol	—	—	Classical aqueous control (in %)
N59C/A295G	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	10%	100.0	20.4	127.0	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose, p-Nitrophenol	—	—	Classical aqueous control (in %)
H8L/N59C/A295G	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Ethanol	10%	100.0	20.4	129.0	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose, p-Nitrophenol	—	—	Classical aqueous control (in %)
H8L	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100.0	74.9	75.3	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose, p-Nitrophenol	—	—	Classical aqueous control (in %)
N59C	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100.0	74.9	101.0	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose, p-Nitrophenol	—	—	Classical aqueous control (in %)
A295G	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100.0	74.9	82.8	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose, p-Nitrophenol	—	—	Classical aqueous control (in %)
N59C/A295G	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100.0	74.9	103.0	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose, p-Nitrophenol	—	—	Classical aqueous control (in %)
H8L/N59C/A295G	Activity - Classical	Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 405 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	10%	100.0	74.9	104.0	%	100 mM sodium phosphate buffer incubation	6	60°C	5 mM p-Nitrophenyl β-D-glucopyranoside	D-Glucose, p-Nitrophenol	—	—	Classical aqueous control (in %)

Mutations in this dataset (6)

WT H8L H8L/N59C/A295G N59C N59C/A295G A295G

Visualization : Activity — Classical

One bar per measurement. Colour = solvent, shade = solvent volume. — — — 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.

Structure

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