Cellulase (gene tox1) from Neurospora crassa

Enzyme Description

Species

Neurospora crassa ↗

Extremophile

EC Number

3.2.1.4 ↗

Sequence

Length: 43 amino acids

MLIYVAEICLSVWLWDGLGDKKKGYAMLGWARPLILGCGSKNQ

Yue Xiao et al. (2016) — Neurospora crassa tox-1 Gene Encodes a pH- and Temperature-Tolerant Mini-Cellulase

Journal of Agricultural and Food Chemistry · doi:10.1021/acs.jafc.6b00043 ↗ · Stability - Incubation

6 measurements ▼

Database ID

UniProt: A0A0K2GUU6 ↗

Sequence Annotation

Explicit - Provided GenBank Accession Number

Protein Source

Recombinant, host bacterium Escherichia coli DH5alpha

Experimental Data (6 measurements)

6 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	Stability - Incubation	Activity after incubation (2 hours at room temperature), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	10%	100.0	—	87.51	%	Incubation: 50 mM phosphate-buffered saline buffer, Assay: 50 mM phosphate-buffered saline buffer	Incubation: 7.4, Assay: 4	Incubation: Room temperature, Assay: 50°C	2.92 mM cellose	Reducing sugars	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Enzyme is a peptide
WT	Stability - Incubation	Activity after incubation (2 hours at room temperature), measured by absorbance spectrophotometry (DNS method, 540 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	100.0	—	96.78	%	Incubation: 50 mM phosphate-buffered saline buffer, Assay: 50 mM phosphate-buffered saline buffer	Incubation: 7.4, Assay: 4	Incubation: Room temperature, Assay: 50°C	2.92 mM cellose	Reducing sugars	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Enzyme is a peptide
Y4L/C9L	Stability - Incubation	Activity after incubation (2 hours at room temperature), measured by absorbance spectrophotometry (DNS method, 540 nm) in aqueous phase	Methanol	10%	100.0	87.51	97.56	%	Incubation: 50 mM phosphate-buffered saline buffer, Assay: 50 mM phosphate-buffered saline buffer	Incubation: 7.4, Assay: 4	Incubation: Room temperature, Assay: 50°C	2.92 mM cellose	Reducing sugars	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Enzyme is a peptide
Y4L/C9L	Stability - Incubation	Activity after incubation (2 hours at room temperature), measured by absorbance spectrophotometry (DNS method, 540 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	100.0	96.78	113.04	%	Incubation: 50 mM phosphate-buffered saline buffer, Assay: 50 mM phosphate-buffered saline buffer	Incubation: 7.4, Assay: 4	Incubation: Room temperature, Assay: 50°C	2.92 mM cellose	Reducing sugars	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Enzyme is a peptide
Y4L/A6L/C9L	Stability - Incubation	Activity after incubation (2 hours at room temperature), measured by absorbance spectrophotometry (DNS method, 540 nm) in aqueous phase	Methanol	10%	100.0	87.51	98.81	%	Incubation: 50 mM phosphate-buffered saline buffer, Assay: 50 mM phosphate-buffered saline buffer	Incubation: 7.4, Assay: 4	Incubation: Room temperature, Assay: 50°C	2.92 mM cellose	Reducing sugars	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Enzyme is a peptide
Y4L/A6L/C9L	Stability - Incubation	Activity after incubation (2 hours at room temperature), measured by absorbance spectrophotometry (DNS method, 540 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	100.0	96.78	94.94	%	Incubation: 50 mM phosphate-buffered saline buffer, Assay: 50 mM phosphate-buffered saline buffer	Incubation: 7.4, Assay: 4	Incubation: Room temperature, Assay: 50°C	2.92 mM cellose	Reducing sugars	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Enzyme is a peptide

WT	Stability - Incubation	Activity after incubation (2 hours at room temperature), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	10%	100.0	—	87.51	%	Incubation: 50 mM phosphate-buffered saline buffer, Assay: 50 mM phosphate-buffered saline buffer	Incubation: 7.4, Assay: 4	Incubation: Room temperature, Assay: 50°C	2.92 mM cellose	Reducing sugars	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Enzyme is a peptide
WT	Stability - Incubation	Activity after incubation (2 hours at room temperature), measured by absorbance spectrophotometry (DNS method, 540 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	100.0	—	96.78	%	Incubation: 50 mM phosphate-buffered saline buffer, Assay: 50 mM phosphate-buffered saline buffer	Incubation: 7.4, Assay: 4	Incubation: Room temperature, Assay: 50°C	2.92 mM cellose	Reducing sugars	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Enzyme is a peptide
Y4L/C9L	Stability - Incubation	Activity after incubation (2 hours at room temperature), measured by absorbance spectrophotometry (DNS method, 540 nm) in aqueous phase	Methanol	10%	100.0	87.51	97.56	%	Incubation: 50 mM phosphate-buffered saline buffer, Assay: 50 mM phosphate-buffered saline buffer	Incubation: 7.4, Assay: 4	Incubation: Room temperature, Assay: 50°C	2.92 mM cellose	Reducing sugars	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Enzyme is a peptide
Y4L/C9L	Stability - Incubation	Activity after incubation (2 hours at room temperature), measured by absorbance spectrophotometry (DNS method, 540 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	100.0	96.78	113.04	%	Incubation: 50 mM phosphate-buffered saline buffer, Assay: 50 mM phosphate-buffered saline buffer	Incubation: 7.4, Assay: 4	Incubation: Room temperature, Assay: 50°C	2.92 mM cellose	Reducing sugars	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Enzyme is a peptide
Y4L/A6L/C9L	Stability - Incubation	Activity after incubation (2 hours at room temperature), measured by absorbance spectrophotometry (DNS method, 540 nm) in aqueous phase	Methanol	10%	100.0	87.51	98.81	%	Incubation: 50 mM phosphate-buffered saline buffer, Assay: 50 mM phosphate-buffered saline buffer	Incubation: 7.4, Assay: 4	Incubation: Room temperature, Assay: 50°C	2.92 mM cellose	Reducing sugars	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Enzyme is a peptide
Y4L/A6L/C9L	Stability - Incubation	Activity after incubation (2 hours at room temperature), measured by absorbance spectrophotometry (DNS method, 540 nm) in aqueous phase	Dimethyl Sulfoxide (DMSO)	10%	100.0	96.78	94.94	%	Incubation: 50 mM phosphate-buffered saline buffer, Assay: 50 mM phosphate-buffered saline buffer	Incubation: 7.4, Assay: 4	Incubation: Room temperature, Assay: 50°C	2.92 mM cellose	Reducing sugars	—	—	Non-incubated control (in %), assay measured in aqueous phase \| Enzyme is a peptide

Mutations in this dataset (3)

WT Y4L/A6L/C9L Y4L/C9L

Visualization : Stability — Incubation

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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