BOKU - Universität für Bodenkultur Wien - Forschungsinformationssystem
Gewählte Publikation:
Ribitsch, D; Heumann, S; Karl, W; Gerlach, J; Leber, R; Birner-Gruenberger, R; Gruber, K; Eiteljoerg, I; Remler, P; Siegert, P; Lange, J; Maurer, KH; Berg, G; Guebitz, GM; Schwab, H.
(2012):
Extracellular serine proteases from Stenotrophomonas maltophilia: Screening, isolation and heterologous expression in E. coli.
J Biotechnol. 2012; 157(1):140-147
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- Abstract:
- 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 degrees 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. (C) 2011 Elsevier B.V. All rights reserved.
- Autor*innen der BOKU Wien:
- Gübitz Georg
- Ribitsch Doris
- Find related publications in this database (using NML MeSH Indexing)
- Alginates - chemistry
- Animals -
- Bacterial Proteins - biosynthesis
- Culture Media - metabolism
- Detergents - chemistry
- Escherichia coli - genetics
- Extracellular Space - metabolism
- Glucuronic Acid - chemistry
- Hexuronic Acids - chemistry
- Insulin - metabolism
- Milk - metabolism
- Molecular Sequence Data -
- Recombinant Proteins - biosynthesis
- Serine Proteases - biosynthesis
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization -
- Stenotrophomonas maltophilia - enzymology
- Find related publications in this database (Keywords)
- Detergent protease
- Stenotrophomonas maltophilia
- Alginate beads
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