Amylopullulanase (gene TbbApu) from Thermoanaerobacter brockii

Enzyme Description

Species
Thermoanaerobacter brockii β†—
Extremophile
Yes thermophilic organism
EC Number
3.2.1.1 β†—, 3.2.1.41 β†—

Sequence

Length: 1674 amino acids
MFKRRTLGFLLSFLLIYTAVFGSMPVQFAKAETDTAPAIANVVGDFQSKIGDSDWNINSDKTVMTYKGNGFYEFTTPVALPAGDYEYKVALNHSWEGGGVPSQGNLSLHLDSDSVVTFYYNYNTSSVTDSTKYTPIPEEKLPRIVGTIQSAIGAGDDWKPETSTAIMRDYKFNNVYEYTANVPKGYYEFKVTLGPSWDINYGLNGEQNGPNIPLNVAYDTKITFYYDSVSHNIWTDYNPPLTGPDNNIYYDDLKHDTHDPFFRSPFGAIKTGDTVTLRIQAKNHDLESAKISYWDDIKKTRTEVPMYKIGQSPDGQYEYWEVKLSFDYPTRIWYYFILKDGTKTAYYGDNDEQLGGVGKATDTVNKDFELTVYDKNLDTPDWMKGAVMYQIFPDRFYNGDPLNDRLKEYSRGFDPVEYHDDWYDLPDNPNDKDKPGYTGDGIWNNDFFGGDLQGINDKLDYLKNLGISVIYLNPIFQSPSNHRYDTTDYTKIDELLGDLDTFKTLMKEAHARGIKVILDGVFNHTSDDSIYFDRYGKYLDNELGAYQAWKQGDQSKSPYGDWYEIKPDGTYEGWWGFDSLPVIRQINGSEYNVKSWADFIINNPNAISKYWLNPDGDKDAGADGWRLDVANEIAHDFWVHFRAAINTVKPNAPMIAELWGDASLDLLGDSFNSVMNYLFRNAVIDFILDKQFDDGNVVHNPIDAAKLDQRLMSIYERYPLPVFYSTMNLLGSHDTMRILTVFGYNSANENQNSQEAKDLAVKRLKLAAILQMGYPGMPSIYYGDEAGQSGGKDPDNRRTFSWGREDKDLQDFFKKVVNIRNENQVLKTGDLETLYANGDVYAFGRRIINGKDVFGNSYPDSVAIVVINKGEAKSVQIDTTKFVRDGVAFTDALSGKTYTVRDGQIVVEVVALDGAILISDPGQNLTAPQPITDLKAVSGNGQVDLSWSAVDRAVSYNIYRSTVKGGLYEKIASNVTQITYIDTDVTNGLKYVYSVTAVDSDGNESALSNEVEAYPAFSIGWAGNMNQVDTHVIGVNNPVEVYAEIWAEGLTDKPGQGENMIAQLGYRYIGDGGQDATRNKVEGVEINKDWTWVDARYVGDSGNNDKYMAKFVPDMVGTWEYIMRFSSNQGQDWTYTKGPDGKTDEAKQFIVVPSNDVEPPTALGLQQPGIESSRVTLNWSLSTDNVAIYGYEIYKSLSETGPFVKIATVADTVYNYVDTDVVNGKVYYYKVVAVDTSFNRTASNIVKATPDIIPIKVIFNVTVPDYTPDDGANIAGNFHDAFWNPSAHQMTKTGPNTYSITLTLNEGTQLEYKYARGSWDKVEKGEYGEEIANRKITVVNQGSNTMVVNDTVQRWRDLPIYIYSPKDNTTVDANTNEIEIKGNTYKGAKVTINDESFVQQENGVFTKVVPLEYGVNTTKIHVEPSGDKNNELTKDITITVIREEPVQEKEPTPTPESEPAPMPEPQPTPTPEPQPSCNYGIVTASTLNLREGASITSKIIGTIPAGKVVKWLEEVNGWYKVDYNGKVGYVSTKYVSSVPDPSKVTVAKSVKVIVKSGLNVRVSSSVAARKIGAVPYGTELKVVEERNGWYLVQYKDGFGYIYSVYTVDTKASVLKTVKVTAKSGLNVRAGDSVKARKIGAVPYGTQLKVVGEYGAWYLIQYKNGFGYVYAKYTK
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
NCBI: MZ322507.1 β†—
Sequence Annotation
Explicit - Provided GenBank Accession Number
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 55.2 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 100.0 120.0 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 100.0 122.0 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 100.0 202.0 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 80.0 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 100.0 100.0 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 80.0 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 100.0 104.7 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 77.3 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 100.0 105.5 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 18.7 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 14.7 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 76.0 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 16.0 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 100.0 141.3 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 100.0 213.3 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 70.7 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 94.7 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 82.4 % Digital Continuous 100 mM potassium phosphate buffer 6.5 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 100.0 112.7 % Digital Continuous 100 mM potassium phosphate buffer 6.5 70Β°C 1 % (v/v) Pullulan Reducing sugars β€” β€” Classical aqueous control (in %)
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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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