Purification and characterization of a novel thermophilic β-galactosidase from Picrophilus torridus of potential industrial application
| Autor: | Jayne Murphy, Gary Walsh |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Hot Temperature
Archaeal Proteins Thermoplasmales Thermoacidophile Microbiology Industrial Microbiology 03 medical and health sciences chemistry.chemical_compound Enzyme Stability 030304 developmental biology chemistry.chemical_classification 0303 health sciences Chromatography Picrophilus torridus biology Molecular mass 030306 microbiology Chromatofocusing Thermophile General Medicine beta-Galactosidase biology.organism_classification Enzyme Isoelectric point chemistry DEAE-Sepharose Molecular Medicine |
| Zdroj: | Extremophiles. 23:783-792 |
| ISSN: | 1433-4909 1431-0651 |
| DOI: | 10.1007/s00792-019-01133-4 |
| Popis: | Intracellular β-galactosidase (E.C 3.2.1.23) produced by the thermoacidophilic archeon Picrophilus torridus DSM 9790 was purified to homogeneity using a combination of DEAE Sepharose, gel filtration, hydroxyapatite and chromatofocusing chromatographies. LC–MS/MS analysis was used to confirm the identity of the purified protein. The enzyme was found to be a homotrimer, with a molecular mass of 157.0 kDa and an isoelectric point of 5.7. To our knowledge, this enzyme has the lowest pH optimum of any intracellular β-galactosidase characterized to date. Maximal activity was exhibited at acidic pH values of 5.0–5.5 and at 70 °C. The enzyme retained > 95% activity after heating to 70 °C for 1 h, or after incubation at pH 5.5 for 1 h. The enzyme may be of interest for high-temperature bioprocessing, such as in the production of lactulose. This investigation suggests that the β-galactosidase activity produced by P. torridus is potentially more useful than several enzymes already characterized for such an application. |
| Databáze: | OpenAIRE |
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