C-terminal truncated (190 last residues) amylopullulanase (gene TbbApu) from Thermoanaerobacter brockii

Enzyme Description

Extremophile
Yes thermophilic organism

Sequence

Length: 1484 amino acids
MFKRRTLGFLLSFLLIYTAVFGSMPVQFAKAETDTAPAIANVVGDFQSKIGDSDWNINSDKTVMTYKGNGFYEFTTPVALPAGDYEYKVALNHSWEGGGVPSQGNLSLHLDSDSVVTFYYNYNTSSVTDSTKYTPIPEEKLPRIVGTIQSAIGAGDDWKPETSTAIMRDYKFNNVYEYTANVPKGYYEFKVTLGPSWDINYGLNGEQNGPNIPLNVAYDTKITFYYDSVSHNIWTDYNPPLTGPDNNIYYDDLKHDTHDPFFRSPFGAIKTGDTVTLRIQAKNHDLESAKISYWDDIKKTRTEVPMYKIGQSPDGQYEYWEVKLSFDYPTRIWYYFILKDGTKTAYYGDNDEQLGGVGKATDTVNKDFELTVYDKNLDTPDWMKGAVMYQIFPDRFYNGDPLNDRLKEYSRGFDPVEYHDDWYDLPDNPNDKDKPGYTGDGIWNNDFFGGDLQGINDKLDYLKNLGISVIYLNPIFQSPSNHRYDTTDYTKIDELLGDLDTFKTLMKEAHARGIKVILDGVFNHTSDDSIYFDRYGKYLDNELGAYQAWKQGDQSKSPYGDWYEIKPDGTYEGWWGFDSLPVIRQINGSEYNVKSWADFIINNPNAISKYWLNPDGDKDAGADGWRLDVANEIAHDFWVHFRAAINTVKPNAPMIAELWGDASLDLLGDSFNSVMNYLFRNAVIDFILDKQFDDGNVVHNPIDAAKLDQRLMSIYERYPLPVFYSTMNLLGSHDTMRILTVFGYNSANENQNSQEAKDLAVKRLKLAAILQMGYPGMPSIYYGDEAGQSGGKDPDNRRTFSWGREDKDLQDFFKKVVNIRNENQVLKTGDLETLYANGDVYAFGRRIINGKDVFGNSYPDSVAIVVINKGEAKSVQIDTTKFVRDGVAFTDALSGKTYTVRDGQIVVEVVALDGAILISDPGQNLTAPQPITDLKAVSGNGQVDLSWSAVDRAVSYNIYRSTVKGGLYEKIASNVTQITYIDTDVTNGLKYVYSVTAVDSDGNESALSNEVEAYPAFSIGWAGNMNQVDTHVIGVNNPVEVYAEIWAEGLTDKPGQGENMIAQLGYRYIGDGGQDATRNKVEGVEINKDWTWVDARYVGDSGNNDKYMAKFVPDMVGTWEYIMRFSSNQGQDWTYTKGPDGKTDEAKQFIVVPSNDVEPPTALGLQQPGIESSRVTLNWSLSTDNVAIYGYEIYKSLSETGPFVKIATVADTVYNYVDTDVVNGKVYYYKVVAVDTSFNRTASNIVKATPDIIPIKVIFNVTVPDYTPDDGANIAGNFHDAFWNPSAHQMTKTGPNTYSITLTLNEGTQLEYKYARGSWDKVEKGEYGEEIANRKITVVNQGSNTMVVNDTVQRWRDLPIYIYSPKDNTTVDANTNEIEIKGNTYKGAKVTINDESFVQQENGVFTKVVPLEYGVNTTKIHVEPSGDKNNELTKDITITVIREEPVQEKEPTPTPESEPAPMPEPQPTPTPEPQPSCNYGIVTA
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 190 residues from NCBI sequence removed)
Protein Source
Recombinant, host bacterium Escherichia coli BL21

Experimental Data (64 measurements)

64 measurements β€” page 1 of 4
Property Assay Solvent Solvent Volume Aqueous Reference Measured Value Units Extraction method Data type 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 1-Butanol 50 % v/v 100.0 176.7 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 Isopropanol 50 % v/v 100.0 272.0 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 % v/v 100.0 152.0 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 % v/v 100.0 338.0 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 % v/v 100.0 128.0 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 % v/v 100.0 225.3 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 % v/v 100.0 126.0 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 % v/v 100.0 232.0 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 % v/v 100.0 92.7 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 Isopropanol 50 % v/v 100.0 102.5 % Digital Continuous 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 % v/v 100.0 30.0 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 % v/v 100.0 20.7 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 % v/v 100.0 110.7 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 % v/v 100.0 28.7 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 % v/v 100.0 176.0 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 % v/v 100.0 340.0 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 % v/v 100.0 100.7 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 50 % v/v 100.0 168.0 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan 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 50 % v/v 100.0 84.0 % Digital Continuous 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 Isopropanol 20 % v/v 100.0 148.7 % Digital Continuous 100 mM potassium phosphate buffer 6 70Β°C 1 % (v/v) Pullulan Reducing sugars β€” β€” Classical aqueous control (in %)
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Visualization : Activity β€” Classical

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Visualization : Stability β€” Incubation

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Structure

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