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RESEARCH PRODUCT
Calculation of the molecular masses of two newly synthesized thermostable enzymes isolated from thermophilic microorganisms
M.h. GeyKlaus K. Ungersubject
chemistry.chemical_classificationHot TemperatureProteaseChromatographybiologyMolecular massmedicine.medical_treatmentThermophileSize-exclusion chromatographyGeneral Chemistrybeta-Galactosidasebiology.organism_classificationBacillalesHigh-performance liquid chromatographyGeobacillus stearothermophilusMolecular WeightEnzymechemistrySephadexEndopeptidasesEnzyme StabilityChromatography GelmedicineChromatography High Pressure Liquiddescription
Two thermostable enzymes synthesized by thermophilic microorganisms were isolated and purified. A thermostable beta-galactosidase was produced in a continuous fermentation process by Bacillus stearothermophilus TP 32 as an intracellular enzyme. After applying different concentration procedures the raw extract enzyme was prepurified on a Sephadex G-200 size exclusion column. The isolated beta-galactosidase fraction was then separated with HPLC on a TSK G 3000 SW size exclusion column to determine the molecular mass based on calibration curves of standard proteins. The other enzyme, a thermostable protease, was synthesized by Bacillus stearothermophilus TP 26 as an extracellular enzyme. After its concentration, the enzyme was purified on a classical size exclusion column (Sephacryl S-200) and on a HPLC size exclusion column (BIO-SIL TSK-250). The micropreparatively isolated fraction was separated again on this HPLC column to determine its molecular mass. The optimum temperature of both enzymes was approximately 75 degrees C.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 1995-04-07 | Journal of Chromatography B: Biomedical Sciences and Applications |