Endo-β-N-acetylglucosaminidase from Arthrobacter protophormiae

Enzyme Description

Species

Extremophile

EC Number

3.2.1.96 ↗, 3.2.1.96 ↗ (hydrolysis)

Sequence

Length: 645 amino acids

MRKAFLVGLVCTACVLLHDDPVAASTYNGPLSSHWFPEELAQWEPDSDPDAPFNRSHVPLEPGRVANRVNANADKDAHLVSLSALNRHTSGVPSQGAPVFYENTFSYWHYTDLMVYWAGSAGEGIIVPPSADVIDASHRNGVPILGNVFFPPTVYGGQLEWLEQMLEQEEDGSFPLADKLLEVADYYGFDGWFINQETEGADEGTAEAMQAFLVYLQEQKPEGMHIMWYDSMIDTGAIAWQNHLTDRNKMYLQNGSTRVADSMFLNFWWRDQRQSNELAQALGRSPYDLYAGVDVEARGTSTPVQWEGLFPEGEKAHTSLGLYRPDWAFQSSETMEAFYEKELQFWVGSTGNPAETDGQSNWPGMAHWFPAKSTATSVPFVTHFNTGSGAQFSAEGKTVSEQEWNNRSLQDVLPTWRWIQHGGDLEATFSWEEAFEGGSSLQWHGSLAEGEHAQIELYQTELPISEGTSLTWTFKSEHGNDLNVGFRLDGEEDFRYVEGEQRESINGWTQWTLPLDAFAGQTITGLAFAAEGNETGLAEFYIGQLAVGADSEKPAAPNVNVRQYDPDPSGIQLVWEKQSNVHHYRVYKETKHGKELIGTSAGDRIYLEGLVEESKQNDVRLHIEALSETFVPSDARMIDIKSGSF

J Q Fan et al. (1995) — Enhanced transglycosylation activity of Arthrobacter protophormiae endo-beta-N-acetylglucosaminidase in media containing organic solvents

Journal of Biological Chemistry · doi:10.1074/jbc.270.30.17723 ↗ · Activity - Classical Activity + Stability - Conversion Specificity - Reaction Specificity

24 measurements ▼

Database ID

UniProt: Q9ZB22 ↗

Sequence Annotation

Inferred - from protein name

Protein Source

Purified, bacterium Arthrobacter protophormiae

Experimental Data (24 measurements)

24 measurements — page 2 of 2

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Specificity - Reaction Specificity	Percentage of transglycosylation product to digested substrate after incubation (15 minutes at 37°C) in the presence of organic solvent, measured by HPLC	Dimethyl Sulfoxide (DMSO)	30%	32.5	98.0	% transglycosylation product	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in % transglycosylation product) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Specificity - Reaction Specificity	Percentage of transglycosylation product to digested substrate after incubation (15 minutes at 37°C) in the presence of organic solvent, measured by HPLC	Ethanol	30%	32.5	98.4	% transglycosylation product	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in % transglycosylation product) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Specificity - Reaction Specificity	Percentage of transglycosylation product to digested substrate after incubation (15 minutes at 37°C) in the presence of organic solvent, measured by HPLC	Methanol	30%	32.5	92.7	% transglycosylation product	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in % transglycosylation product) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Specificity - Reaction Specificity	Percentage of transglycosylation product to digested substrate after incubation (15 minutes at 37°C) in the presence of organic solvent, measured by HPLC	Tetrahydrofuran (THF)	30%	32.5	0.0	% transglycosylation product	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in % transglycosylation product) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)

Activity - Classical	Activity measured by mass spectrometry in the presence of organic solvent	Acetonitrile	30%	100.0	6.9	%	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre Man9GlcNAc2-Asn	GlcNAc–Asn, Man9GlcNAc	—	—	Classical aqueous control (in %) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Activity - Classical	Activity measured by mass spectrometry in the presence of organic solvent	1,4-Dioxane	30%	100.0	79.3	%	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre Man9GlcNAc2-Asn	GlcNAc–Asn, Man9GlcNAc	—	—	Classical aqueous control (in %) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Activity - Classical	Activity measured by mass spectrometry in the presence of organic solvent	Dimethylformamide (DMF)	30%	100.0	64.2	%	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre Man9GlcNAc2-Asn	GlcNAc–Asn, Man9GlcNAc	—	—	Classical aqueous control (in %) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Activity - Classical	Activity measured by mass spectrometry in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	30%	100.0	99.0	%	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre Man9GlcNAc2-Asn	GlcNAc–Asn, Man9GlcNAc	—	—	Classical aqueous control (in %) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Activity - Classical	Activity measured by mass spectrometry in the presence of organic solvent	Ethanol	30%	100.0	55.0	%	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre Man9GlcNAc2-Asn	GlcNAc–Asn, Man9GlcNAc	—	—	Classical aqueous control (in %) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Activity - Classical	Activity measured by mass spectrometry in the presence of organic solvent	Methanol	30%	100.0	32.5	%	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre Man9GlcNAc2-Asn	GlcNAc–Asn, Man9GlcNAc	—	—	Classical aqueous control (in %) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Activity - Classical	Activity measured by mass spectrometry in the presence of organic solvent	Tetrahydrofuran (THF)	30%	100.0	9.2	%	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre Man9GlcNAc2-Asn	GlcNAc–Asn, Man9GlcNAc	—	—	Classical aqueous control (in %) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Activity - Classical	Activity measured by mass spectrometry in the presence of organic solvent	Acetone	30%	100.0	100.0	%	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre Man9GlcNAc2-Asn	GlcNAc–Asn, Man9GlcNAc	—	—	Classical aqueous control (in %) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Activity + Stability - Conversion	Conversion measured by HPLC after incubation (15 minutes at 37°C) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	30%	98.0	75.2	%	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	GlcNAc–Asn, Man9GlcNAc, Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in %) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)/Both hydrolysis and transglycosylation reactions measured
Activity + Stability - Conversion	Conversion measured by HPLC after incubation (15 minutes at 37°C) in the presence of organic solvent	Tetrahydrofuran (THF)	30%	98.0	1.4	%	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	GlcNAc–Asn, Man9GlcNAc, Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in %) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)/Both hydrolysis and transglycosylation reactions measured
Activity + Stability - Conversion	Conversion measured by HPLC after incubation (15 minutes at 37°C) in the presence of organic solvent	Methanol	30%	98.0	97.5	%	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	GlcNAc–Asn, Man9GlcNAc, Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in %) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)/Both hydrolysis and transglycosylation reactions measured
Activity + Stability - Conversion	Conversion measured by HPLC after incubation (15 minutes at 37°C) in the presence of organic solvent	Ethanol	30%	98.0	52.9	%	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	GlcNAc–Asn, Man9GlcNAc, Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in %) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)/Both hydrolysis and transglycosylation reactions measured
Activity + Stability - Conversion	Conversion measured by HPLC after incubation (15 minutes at 37°C) in the presence of organic solvent	Dimethylformamide (DMF)	30%	98.0	28.3	%	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	GlcNAc–Asn, Man9GlcNAc, Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in %) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)/Both hydrolysis and transglycosylation reactions measured
Activity + Stability - Conversion	Conversion measured by HPLC after incubation (15 minutes at 37°C) in the presence of organic solvent	1,4-Dioxane	30%	98.0	14.4	%	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	GlcNAc–Asn, Man9GlcNAc, Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in %) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)/Both hydrolysis and transglycosylation reactions measured
Activity + Stability - Conversion	Conversion measured by HPLC after incubation (15 minutes at 37°C) in the presence of organic solvent	Acetonitrile	30%	98.0	0.02	%	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	GlcNAc–Asn, Man9GlcNAc, Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in %) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)/Both hydrolysis and transglycosylation reactions measured
Activity + Stability - Conversion	Conversion measured by HPLC after incubation (15 minutes at 37°C) in the presence of organic solvent	Acetone	30%	98.0	91.9	%	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	GlcNAc–Asn, Man9GlcNAc, Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in %) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)/Both hydrolysis and transglycosylation reactions measured
Specificity - Reaction Specificity	Percentage of transglycosylation product to digested substrate after incubation (15 minutes at 37°C) in the presence of organic solvent, measured by HPLC	Acetone	30%	32.5	99.1	% transglycosylation product	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in % transglycosylation product) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Specificity - Reaction Specificity	Percentage of transglycosylation product to digested substrate after incubation (15 minutes at 37°C) in the presence of organic solvent, measured by HPLC	Acetonitrile	30%	32.5	100.0	% transglycosylation product	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in % transglycosylation product) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Specificity - Reaction Specificity	Percentage of transglycosylation product to digested substrate after incubation (15 minutes at 37°C) in the presence of organic solvent, measured by HPLC	1,4-Dioxane	30%	32.5	97.0	% transglycosylation product	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in % transglycosylation product) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Specificity - Reaction Specificity	Percentage of transglycosylation product to digested substrate after incubation (15 minutes at 37°C) in the presence of organic solvent, measured by HPLC	Dimethylformamide (DMF)	30%	32.5	100.0	% transglycosylation product	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in % transglycosylation product) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Specificity - Reaction Specificity	Percentage of transglycosylation product to digested substrate after incubation (15 minutes at 37°C) in the presence of organic solvent, measured by HPLC	Dimethyl Sulfoxide (DMSO)	30%	32.5	98.0	% transglycosylation product	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in % transglycosylation product) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Specificity - Reaction Specificity	Percentage of transglycosylation product to digested substrate after incubation (15 minutes at 37°C) in the presence of organic solvent, measured by HPLC	Ethanol	30%	32.5	98.4	% transglycosylation product	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in % transglycosylation product) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Specificity - Reaction Specificity	Percentage of transglycosylation product to digested substrate after incubation (15 minutes at 37°C) in the presence of organic solvent, measured by HPLC	Methanol	30%	32.5	92.7	% transglycosylation product	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in % transglycosylation product) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)
Specificity - Reaction Specificity	Percentage of transglycosylation product to digested substrate after incubation (15 minutes at 37°C) in the presence of organic solvent, measured by HPLC	Tetrahydrofuran (THF)	30%	32.5	0.0	% transglycosylation product	25 mM ammonium acetate buffer	6	37°C	3 nmol/20microlitre of Man9GlcNAc2Asn, N-Acetyl-D-Glucosamine	Man9GlcNAc–GlcNAc	—	—	Classical aqueous control (in % transglycosylation product) \| Same organism as Glutamicibacter protophormiae (Brevibacterium protophormiae)

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Visualization : Activity — Classical

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

Visualization : Activity + Stability — Conversion

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

Visualization : Specificity — Reaction Specificity

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

Structure

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Endo-β-N-acetylglucosaminidase from Arthrobacter protophormiae

Enzyme Description

Sequence

Experimental Data (24 measurements)

Visualization : Activity — Classical

Visualization : Activity + Stability — Conversion

Visualization : Specificity — Reaction Specificity

Structure

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