title: Calculation of the molecular masses of two newly synthesized thermostable enzymes isolated from thermophilic microorganisms.
abstract: 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.
authors.affiliation: Institute of Anorganic and Analytical Chemistry, Johannes Gutenberg University Mainz, Germany.
authors.forename: M H
authors.initials: MH
authors.lastname: Gey
authors.forename: K K
authors.initials: KK
authors.lastname: Unger
mesh_terms: D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D010450:Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification; D004795:Enzyme Stability; D001411:Geobacillus stearothermophilus / Q000201:enzymology*; D006358:Hot Temperature; D008970:Molecular Weight; D001616:beta-Galactosidase / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D001616:beta-Galactosidase; D010450:Endopeptidases