C-terminal truncated (420 last residues) amylopullulanase (gene TbbApu) from Thermoanaerobacter brockii

Enzyme Description

Extremophile
Yes thermophilic organism

Sequence

Length: 1254 amino acids
MFKRRTLGFLLSFLLIYTAVFGSMPVQFAKAETDTAPAIANVVGDFQSKIGDSDWNINSDKTVMTYKGNGFYEFTTPVALPAGDYEYKVALNHSWEGGGVPSQGNLSLHLDSDSVVTFYYNYNTSSVTDSTKYTPIPEEKLPRIVGTIQSAIGAGDDWKPETSTAIMRDYKFNNVYEYTANVPKGYYEFKVTLGPSWDINYGLNGEQNGPNIPLNVAYDTKITFYYDSVSHNIWTDYNPPLTGPDNNIYYDDLKHDTHDPFFRSPFGAIKTGDTVTLRIQAKNHDLESAKISYWDDIKKTRTEVPMYKIGQSPDGQYEYWEVKLSFDYPTRIWYYFILKDGTKTAYYGDNDEQLGGVGKATDTVNKDFELTVYDKNLDTPDWMKGAVMYQIFPDRFYNGDPLNDRLKEYSRGFDPVEYHDDWYDLPDNPNDKDKPGYTGDGIWNNDFFGGDLQGINDKLDYLKNLGISVIYLNPIFQSPSNHRYDTTDYTKIDELLGDLDTFKTLMKEAHARGIKVILDGVFNHTSDDSIYFDRYGKYLDNELGAYQAWKQGDQSKSPYGDWYEIKPDGTYEGWWGFDSLPVIRQINGSEYNVKSWADFIINNPNAISKYWLNPDGDKDAGADGWRLDVANEIAHDFWVHFRAAINTVKPNAPMIAELWGDASLDLLGDSFNSVMNYLFRNAVIDFILDKQFDDGNVVHNPIDAAKLDQRLMSIYERYPLPVFYSTMNLLGSHDTMRILTVFGYNSANENQNSQEAKDLAVKRLKLAAILQMGYPGMPSIYYGDEAGQSGGKDPDNRRTFSWGREDKDLQDFFKKVVNIRNENQVLKTGDLETLYANGDVYAFGRRIINGKDVFGNSYPDSVAIVVINKGEAKSVQIDTTKFVRDGVAFTDALSGKTYTVRDGQIVVEVVALDGAILISDPGQNLTAPQPITDLKAVSGNGQVDLSWSAVDRAVSYNIYRSTVKGGLYEKIASNVTQITYIDTDVTNGLKYVYSVTAVDSDGNESALSNEVEAYPAFSIGWAGNMNQVDTHVIGVNNPVEVYAEIWAEGLTDKPGQGENMIAQLGYRYIGDGGQDATRNKVEGVEINKDWTWVDARYVGDSGNNDKYMAKFVPDMVGTWEYIMRFSSNQGQDWTYTKGPDGKTDEAKQFIVVPSNDVEPPTALGLQQPGIESSRVTLNWSLSTDNVAIYGYEIYKSLSETGPFVKIATVADTVYNYVDTDVVNGKVYYYKVVAVDTSFNRTASNIVKATPDIIP
Hande Mumcu et al. (2023) β€” Cloning and characterization of thermostable amylopullulanase TbbApu and its C-terminal truncated variants with enhanced activity in organic solvents
Enzyme and Microbial Technology  Β· doi:10.1016/j.enzmictec.2022.110176 β†—  Β· Activity - Classical Stability - Incubation
64 measurements
Database ID
Sequence Annotation
Explicit - Provided GenBank Accession Number (last 420 residues from NCBI sequence removed)
Protein Source
Recombinant, host bacterium Escherichia coli BL21

Experimental Data (64 measurements)

64 measurements β€” page 2 of 4
Property Assay Solvent Solvent Volume Aqueous Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
Activity - Classical Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent Isopropanol 20% 100.0 190.8 % 100 mM potassium phosphate buffer 6 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” Classical aqueous control (in %)
Activity - Classical Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent Acetone 20% 100.0 195.8 % 100 mM potassium phosphate buffer 6 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” Classical aqueous control (in %)
Activity - Classical Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent Acetone 50% 100.0 179.8 % 100 mM potassium phosphate buffer 6 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” Classical aqueous control (in %)
Activity - Classical Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent Ethanol 20% 100.0 132.3 % 100 mM potassium phosphate buffer 6 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” 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 89.5 % 100 mM potassium phosphate buffer 6 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” Classical aqueous control (in %)
Activity - Classical Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent Methanol 20% 100.0 125.6 % 100 mM potassium phosphate buffer 6 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” Classical aqueous control (in %)
Activity - Classical Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent Methanol 50% 100.0 119.7 % 100 mM potassium phosphate buffer 6 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” 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 39.1 % 100 mM potassium phosphate buffer 6 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” 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 13.9 % 100 mM potassium phosphate buffer 6 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” 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 25.2 % 100 mM potassium phosphate buffer 6 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” 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 13.0 % 100 mM potassium phosphate buffer 6 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” 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 22.3 % 100 mM potassium phosphate buffer 6 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” Classical aqueous control (in %)
Activity - Classical Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent n-Hexane 20% 100.0 133.6 % 100 mM potassium phosphate buffer 6 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” Classical aqueous control (in %)
Activity - Classical Activity measured by absorbance spectrophotometry (DNS assay, 540 nm) in the presence of organic solvent n-Hexane 50% 100.0 204.6 % 100 mM potassium phosphate buffer 6 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” 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 78.2 % 100 mM potassium phosphate buffer 6 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” 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 77.3 % 100 mM potassium phosphate buffer 6 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” Classical aqueous control (in %)
Stability - Incubation Activity after incubation (24 hours at 25Β°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase Isopropanol 50% 100.0 51.0 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 6 Incubation: 25Β°C, Assay: 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” 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 25Β°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 Incubation: 25Β°C, Assay: 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” 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 25Β°C), measured by absorbance spectrophotometry (DNS assay, 540 nm) in aqueous phase Ethanol 50% 100.0 28.3 % Incubation: ?, Assay: 100 mM potassium phosphate buffer Incubation: ?, Assay: 6 Incubation: 25Β°C, Assay: 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” 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 25Β°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 Incubation: 25Β°C, Assay: 75Β°C 1 % (v/v) Starch Reducing sugars β€” β€” 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 β€” Classical

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

Visualization : Stability β€” Incubation

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

Structure

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