Extracellular serine proteases from Stenotrophomonas maltophilia: Screening, isolation and heterologous expression in E. coli
| Autor: | Doris Ribitsch, R. Leber, Petra Siegert, Ruth Birner-Gruenberger, Inge Eiteljoerg, J. Lange, Wolfgang Karl, Karl Gruber, Jochen Prof. Dr. Gerlach, Georg M. Guebitz, Helmut Schwab, P. Remler, Sonja Heumann, K.H. Maurer, Gabriele Berg |
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| Rok vydání: | 2011 |
| Předmět: |
Proteases
Alginates medicine.medical_treatment Stenotrophomonas maltophilia Detergents Molecular Sequence Data Bioengineering Biology Applied Microbiology and Biotechnology Microbiology Serine Bacterial Proteins Glucuronic Acid Extracellular medicine Escherichia coli Animals Insulin chemistry.chemical_classification Protease Hexuronic Acids General Medicine biology.organism_classification Recombinant Proteins Culture Media Enzyme Milk Biochemistry chemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Heterologous expression Serine Proteases Extracellular Space Bacteria Biotechnology |
| Zdroj: | Journal of biotechnology. 157(1) |
| ISSN: | 1873-4863 |
| Popis: | A large strain collection comprising antagonistic bacteria was screened for novel detergent proteases. Several strains displayed protease activity on agar plates containing skim milk but were inactive in liquid media. Encapsulation of cells in alginate beads induced protease production. Stenotrophomonas maltophilia emerged as best performer under washing conditions. For identification of wash-active proteases, four extracellular serine proteases called StmPr1, StmPr2, StmPr3 and StmPr4 were cloned. StmPr2 and StmPr4 were sufficiently overexpressed in E. coli . Expression of StmPr1 and StmPr3 resulted in unprocessed, insoluble protein. Truncation of most of the C-terminal domain which has been identified by enzyme modeling succeeded in expression of soluble, active StmPr1 but failed in case of StmPr3. From laundry application tests StmPr2 turned out to be a highly wash-active protease at 45 °C. Specific activity of StmPr2 determined with suc - l -Ala- l -Ala- l -Pro- l -Phe- p -nitroanilide as the substrate was 17 ± 2 U/mg. In addition we determined the kinetic parameters and cleavage preferences of protease StmPr2. |
| Databáze: | OpenAIRE |
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