Amylopullulanase (gene TbbApu) from Thermoanaerobacter brockii

Enzyme Description

Species

Thermoanaerobacter brockii ↗

Extremophile

Yes thermophilic organism

EC Number

3.2.1.1 ↗, 3.2.1.41 ↗

Sequence

Length: 1674 amino acids

MFKRRTLGFLLSFLLIYTAVFGSMPVQFAKAETDTAPAIANVVGDFQSKIGDSDWNINSDKTVMTYKGNGFYEFTTPVALPAGDYEYKVALNHSWEGGGVPSQGNLSLHLDSDSVVTFYYNYNTSSVTDSTKYTPIPEEKLPRIVGTIQSAIGAGDDWKPETSTAIMRDYKFNNVYEYTANVPKGYYEFKVTLGPSWDINYGLNGEQNGPNIPLNVAYDTKITFYYDSVSHNIWTDYNPPLTGPDNNIYYDDLKHDTHDPFFRSPFGAIKTGDTVTLRIQAKNHDLESAKISYWDDIKKTRTEVPMYKIGQSPDGQYEYWEVKLSFDYPTRIWYYFILKDGTKTAYYGDNDEQLGGVGKATDTVNKDFELTVYDKNLDTPDWMKGAVMYQIFPDRFYNGDPLNDRLKEYSRGFDPVEYHDDWYDLPDNPNDKDKPGYTGDGIWNNDFFGGDLQGINDKLDYLKNLGISVIYLNPIFQSPSNHRYDTTDYTKIDELLGDLDTFKTLMKEAHARGIKVILDGVFNHTSDDSIYFDRYGKYLDNELGAYQAWKQGDQSKSPYGDWYEIKPDGTYEGWWGFDSLPVIRQINGSEYNVKSWADFIINNPNAISKYWLNPDGDKDAGADGWRLDVANEIAHDFWVHFRAAINTVKPNAPMIAELWGDASLDLLGDSFNSVMNYLFRNAVIDFILDKQFDDGNVVHNPIDAAKLDQRLMSIYERYPLPVFYSTMNLLGSHDTMRILTVFGYNSANENQNSQEAKDLAVKRLKLAAILQMGYPGMPSIYYGDEAGQSGGKDPDNRRTFSWGREDKDLQDFFKKVVNIRNENQVLKTGDLETLYANGDVYAFGRRIINGKDVFGNSYPDSVAIVVINKGEAKSVQIDTTKFVRDGVAFTDALSGKTYTVRDGQIVVEVVALDGAILISDPGQNLTAPQPITDLKAVSGNGQVDLSWSAVDRAVSYNIYRSTVKGGLYEKIASNVTQITYIDTDVTNGLKYVYSVTAVDSDGNESALSNEVEAYPAFSIGWAGNMNQVDTHVIGVNNPVEVYAEIWAEGLTDKPGQGENMIAQLGYRYIGDGGQDATRNKVEGVEINKDWTWVDARYVGDSGNNDKYMAKFVPDMVGTWEYIMRFSSNQGQDWTYTKGPDGKTDEAKQFIVVPSNDVEPPTALGLQQPGIESSRVTLNWSLSTDNVAIYGYEIYKSLSETGPFVKIATVADTVYNYVDTDVVNGKVYYYKVVAVDTSFNRTASNIVKATPDIIPIKVIFNVTVPDYTPDDGANIAGNFHDAFWNPSAHQMTKTGPNTYSITLTLNEGTQLEYKYARGSWDKVEKGEYGEEIANRKITVVNQGSNTMVVNDTVQRWRDLPIYIYSPKDNTTVDANTNEIEIKGNTYKGAKVTINDESFVQQENGVFTKVVPLEYGVNTTKIHVEPSGDKNNELTKDITITVIREEPVQEKEPTPTPESEPAPMPEPQPTPTPEPQPSCNYGIVTASTLNLREGASITSKIIGTIPAGKVVKWLEEVNGWYKVDYNGKVGYVSTKYVSSVPDPSKVTVAKSVKVIVKSGLNVRVSSSVAARKIGAVPYGTELKVVEERNGWYLVQYKDGFGYIYSVYTVDTKASVLKTVKVTAKSGLNVRAGDSVKARKIGAVPYGTQLKVVGEYGAWYLIQYKNGFGYVYAKYTK

Hande Mumcu et al. (2023) — Cloning and characterization of thermostable amylopullulanase TbbApu and its C-terminal truncated variants with enhanced activity in organic solvents

Enzyme and Microbial Technology · doi:10.1016/j.enzmictec.2022.110176 ↗ · Activity - Classical Stability - Incubation

64 measurements ▼

Database ID

NCBI: MZ322507.1 ↗

Sequence Annotation

Explicit - Provided GenBank Accession Number

Protein Source

Recombinant, host bacterium Escherichia coli BL21

Experimental Data (64 measurements)

64 measurements — page 3 of 4

Property	Assay	Solvent	Solvent Volume	Aqueous Reference	Measured Value	Units	Solution	pH	Temperature	Substrate(s)	Product(s)	Cofactor(s)	Shaking	Comments
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	1-Butanol	50%	100.0	38.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	n-Hexane	50%	100.0	88.6	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Glycerol	50%	100.0	18.1	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Isopropanol	50%	100.0	36.1	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Acetone	50%	100.0	90.7	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Ethanol	50%	100.0	38.8	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	50%	100.0	41.8	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	1-Butanol	50%	100.0	30.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	n-Hexane	50%	100.0	100.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Glycerol	50%	100.0	39.6	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Glycerol	20%	100.0	55.8	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	n-Hexane	20%	100.0	100.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	1-Butanol	20%	100.0	48.1	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	20%	100.0	76.8	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Ethanol	20%	100.0	57.9	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Acetone	20%	100.0	92.3	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Isopropanol	20%	100.0	66.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Glycerol	20%	100.0	53.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	n-Hexane	20%	100.0	100.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	1-Butanol	20%	100.0	47.9	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")

Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	1-Butanol	50%	100.0	55.2	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Isopropanol	50%	100.0	120.0	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Acetone	20%	100.0	122.0	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Acetone	50%	100.0	202.0	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Ethanol	20%	100.0	80.0	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Ethanol	50%	100.0	100.0	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Methanol	20%	100.0	80.0	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Methanol	50%	100.0	104.7	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	1-Butanol	20%	100.0	77.3	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Isopropanol	50%	100.0	105.5	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Dimethylformamide (DMF)	20%	100.0	18.7	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Dimethylformamide (DMF)	50%	100.0	14.7	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	20%	100.0	76.0	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	50%	100.0	16.0	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	n-Hexane	20%	100.0	141.3	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	n-Hexane	50%	100.0	213.3	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Glycerol	20%	100.0	70.7	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Glycerol	50%	100.0	94.7	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Glycerol	50%	100.0	82.4	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Isopropanol	20%	100.0	112.7	%	100 mM potassium phosphate buffer	6.5	70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Isopropanol	20%	100.0	100.4	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Acetone	20%	100.0	94.1	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Acetone	50%	100.0	116.4	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Ethanol	20%	100.0	83.2	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Ethanol	50%	100.0	61.3	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Methanol	20%	100.0	73.1	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Methanol	50%	100.0	89.1	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	1-Butanol	20%	100.0	41.2	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Dimethylformamide (DMF)	20%	100.0	28.2	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Dimethylformamide (DMF)	50%	100.0	29.4	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	20%	100.0	39.1	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Dimethyl Sulfoxide (DMSO)	50%	100.0	53.4	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	n-Hexane	20%	100.0	93.3	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	n-Hexane	50%	100.0	116.4	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	Glycerol	20%	100.0	73.5	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Activity - Classical	Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent	1-Butanol	50%	100.0	49.2	%	100 mM potassium phosphate buffer	6.5	75°C	1 % (v/v) Starch	Reducing sugars	—	—	Classical aqueous control (in %)
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Isopropanol	50%	100.0	56.1	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Acetone	50%	100.0	100.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Ethanol	50%	100.0	34.2	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	50%	100.0	35.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	1-Butanol	50%	100.0	38.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	n-Hexane	50%	100.0	88.6	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Glycerol	50%	100.0	18.1	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Isopropanol	50%	100.0	36.1	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Acetone	50%	100.0	90.7	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Ethanol	50%	100.0	38.8	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	50%	100.0	41.8	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	1-Butanol	50%	100.0	30.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	n-Hexane	50%	100.0	100.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Glycerol	50%	100.0	39.6	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Glycerol	20%	100.0	55.8	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	n-Hexane	20%	100.0	100.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	1-Butanol	20%	100.0	48.1	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	20%	100.0	76.8	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Ethanol	20%	100.0	57.9	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Acetone	20%	100.0	92.3	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Isopropanol	20%	100.0	66.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 70°C	1 % (v/v) Pullulan	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Glycerol	20%	100.0	53.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	n-Hexane	20%	100.0	100.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	1-Butanol	20%	100.0	47.9	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Methanol	20%	100.0	53.0	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Ethanol	20%	100.0	67.8	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Acetone	20%	100.0	97.9	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")
Stability - Incubation	Activity after incubation (24 hours at 25°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase	Isopropanol	20%	100.0	58.9	%	Incubation: ?, Assay: 100 mM potassium phosphate buffer	Incubation: ?, Assay: 6.5	Incubation: 25°C, Assay: 75°C	1 % (v/v) Starch	Reducing sugars	—	—	Non-incubated control (in %), assay in aqueous phase \| Clear about assay in diluted aqueous conditions ("organic solvent treated samples were diluted at a 1:3 ratio and a standard activity assay was performed for each enzyme")

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Visualization : Activity — Classical

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

Visualization : Stability — Incubation