Ξ±-amylase from Brachybacterium sp. LB25
Enzyme Description
Sequence
MQHSPSGTAAPRPLIRLAAALALALLLAVAGSPATSPPAQGSPPVPSGGATSENGRGDAILNLFQWTWDSVAAECTSTIGPAGFGYVQVSPPQETIQGTAWWTSYQPVSYKIEGKLGTRAEFAAMVETCDAAGVEVIVDAVINHTTGADGGSGTGTAGTPYGIDDFPGIYGAADFNDCRTDISSYQDRYQVQNCRLLSLQDLRTGSEYVQNTLAGYMNDLLSLGVAGFRIDAAKHIPASDLAAIKAKLSDPNAYWVQEVIGAAGEPVQVSEYTGTGDVHEFDYARQLKSDFDGSIANLQRISNGKLPSAQAGVFVDNHDTERNGETMNETWGAKYVLANGETMNETWGAKYVLANTFMLAYPYGSPSVYSGYTFTDEDAGAPGATATSVPDANCDSDAWTCTQRWTEIQGMVGFYNTVAGTALTNWWDDGGNQIAFGRGDKGFVAINNTDSATTRTYSSSLPAGTYCDVVAASDCSETITVAADGTFSATLPAYGALALHVEKTSGGTTPDPDPGTDPVAGKVSASVHASTVLGQEVRIVGSVPELGGWQPASGVALDASGYPSWTGGVDLPAGTSFEYKYVKVDDSGAVVWESGANRTATVGADGTLALNDTWR
Noriyuki Doukyu et al. (2007)
β Purification and characterization of a maltooligosaccharide-forming amylase that improves product selectivity in water-miscible organic solvents, from dimethylsulfoxide-tolerant Brachybacterium sp. strain LB25
Extremophiles
Β· doi:10.1007/s00792-007-0096-8 β
Β· Activity - Classical Activity + Stability - Product Formation
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Database ID
UniProt: Q1JUA3 β
Sequence Annotation
Inferred - from protein name
Protein Source
Purified, bacterium Brachybacterium sp. LB25
Experimental Data (17 measurements)
17 measurements
| 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 method, 540 nm) in the presence of organic solvent | Dimethylformamide (DMF) | 20 % v/v | 100.0 | 78.7 | % | Direct | Continuous | 100 mM PIPES-NaOH | 7 | 30Β°C | 0.5% (wt/vol) Starch | Reducing sugars | β | Yes, "the mixture was shaken" | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (DNS method, 540 nm) in the presence of organic solvent | Cyclohexane | 20 % v/v | 100.0 | 94.2 | % | Direct | Continuous | 100 mM PIPES-NaOH | 7 | 30Β°C | 0.5% (wt/vol) Starch | Reducing sugars | β | Yes, "the mixture was shaken" | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (DNS method, 540 nm) in the presence of organic solvent | Toluene | 20 % v/v | 100.0 | 91.6 | % | Direct | Continuous | 100 mM PIPES-NaOH | 7 | 30Β°C | 0.5% (wt/vol) Starch | Reducing sugars | β | Yes, "the mixture was shaken" | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (DNS method, 540 nm) in the presence of organic solvent | Benzene | 20 % v/v | 100.0 | 90.5 | % | Direct | Continuous | 100 mM PIPES-NaOH | 7 | 30Β°C | 0.5% (wt/vol) Starch | Reducing sugars | β | Yes, "the mixture was shaken" | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (DNS method, 540 nm) in the presence of organic solvent | Chloroform | 20 % v/v | 100.0 | 82.6 | % | Direct | Continuous | 100 mM PIPES-NaOH | 7 | 30Β°C | 0.5% (wt/vol) Starch | Reducing sugars | β | Yes, "the mixture was shaken" | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (DNS method, 540 nm) in the presence of organic solvent | Isopropanol | 20 % v/v | 100.0 | 69.2 | % | Direct | Continuous | 100 mM PIPES-NaOH | 7 | 30Β°C | 0.5% (wt/vol) Starch | Reducing sugars | β | Yes, "the mixture was shaken" | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (DNS method, 540 nm) in the presence of organic solvent | Ethanol | 20 % v/v | 100.0 | 72.9 | % | Direct | Continuous | 100 mM PIPES-NaOH | 7 | 30Β°C | 0.5% (wt/vol) Starch | Reducing sugars | β | Yes, "the mixture was shaken" | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (DNS method, 540 nm) in the presence of organic solvent | Acetone | 20 % v/v | 100.0 | 68.5 | % | Direct | Continuous | 100 mM PIPES-NaOH | 7 | 30Β°C | 0.5% (wt/vol) Starch | Reducing sugars | β | Yes, "the mixture was shaken" | Classical aqueous control (in %) |
| Activity - Classical | Activity measured by absorbance spectrophotometry (DNS method, 540 nm) in the presence of organic solvent | Dimethyl Sulfoxide (DMSO) | 20 % v/v | 100.0 | 85.8 | % | Direct | Continuous | 100 mM PIPES-NaOH | 7 | 30Β°C | 0.5% (wt/vol) Starch | Reducing sugars | β | Yes, "the mixture was shaken" | Classical aqueous control (in %) |
| Activity + Stability - Product Formation | Total yield in maltooligosaccharides after incubation (6 hours at 30Β°C) in the presence of organic solvent, measured by HPLC | Dimethyl Sulfoxide (DMSO) | 5 % v/v | 58.7 | 56.0 | % | Direct | Continuous | 100 mM sodium phosphate buffer | 7 | 30Β°C | 1%(wt/vol) Starch | Maltooligosaccharides | β | β | Classical aqueous control (in %) |
| Activity + Stability - Product Formation | Total yield in maltooligosaccharides after incubation (6 hours at 30Β°C) in the presence of organic solvent, measured by HPLC | Dimethyl Sulfoxide (DMSO) | 10 % v/v | 58.7 | 56.8 | % | Direct | Continuous | 100 mM sodium phosphate buffer | 7 | 30Β°C | 1%(wt/vol) Starch | Maltooligosaccharides | β | β | Classical aqueous control (in %) |
| Activity + Stability - Product Formation | Total yield in maltooligosaccharides after incubation (6 hours at 30Β°C) in the presence of organic solvent, measured by HPLC | Dimethyl Sulfoxide (DMSO) | 20 % v/v | 58.7 | 54.4 | % | Direct | Continuous | 100 mM sodium phosphate buffer | 7 | 30Β°C | 1%(wt/vol) Starch | Maltooligosaccharides | β | β | Classical aqueous control (in %) |
| Activity + Stability - Product Formation | Total yield in maltooligosaccharides after incubation (6 hours at 30Β°C) in the presence of organic solvent, measured by HPLC | Dimethyl Sulfoxide (DMSO) | 30 % v/v | 58.7 | 48.9 | % | Direct | Continuous | 100 mM sodium phosphate buffer | 7 | 30Β°C | 1%(wt/vol) Starch | Maltooligosaccharides | β | β | Classical aqueous control (in %) |
| Activity + Stability - Product Formation | Total yield in maltooligosaccharides after incubation (6 hours at 30Β°C) in the presence of organic solvent, measured by HPLC | Ethanol | 5 % v/v | 58.7 | 57.5 | % | Direct | Continuous | 100 mM sodium phosphate buffer | 7 | 30Β°C | 1%(wt/vol) Starch | Maltooligosaccharides | β | β | Classical aqueous control (in %) |
| Activity + Stability - Product Formation | Total yield in maltooligosaccharides after incubation (6 hours at 30Β°C) in the presence of organic solvent, measured by HPLC | Ethanol | 10 % v/v | 58.7 | 57.6 | % | Direct | Continuous | 100 mM sodium phosphate buffer | 7 | 30Β°C | 1%(wt/vol) Starch | Maltooligosaccharides | β | β | Classical aqueous control (in %) |
| Activity + Stability - Product Formation | Total yield in maltooligosaccharides after incubation (6 hours at 30Β°C) in the presence of organic solvent, measured by HPLC | Ethanol | 20 % v/v | 58.7 | 52.9 | % | Direct | Continuous | 100 mM sodium phosphate buffer | 7 | 30Β°C | 1%(wt/vol) Starch | Maltooligosaccharides | β | β | Classical aqueous control (in %) |
| Activity + Stability - Product Formation | Total yield in maltooligosaccharides after incubation (6 hours at 30Β°C) in the presence of organic solvent, measured by HPLC | Ethanol | 30 % v/v | 58.7 | 27.8 | % | Direct | Continuous | 100 mM sodium phosphate buffer | 7 | 30Β°C | 1%(wt/vol) Starch | Maltooligosaccharides | β | β | Classical aqueous control (in %) |
Visualization : Activity β Classical
One bar per measurement. Colour = solvent, shade = solvent volume.
Visualization : Activity + Stability β Product Formation
One bar per measurement. Colour = solvent, shade = solvent volume.