Analysis of secreted protein profile and enzymatic activities from Corynebacterium diphtheriae and Bordetella pertussis on production batch media using peptide quenched fluorescent substrates
| Autor: | Maria Aparecida Sakauchi, Ivo Lebrun, Sally M. A. Prado, L. Juliano, Elen Aquino Perpetuo, Maria A. Juliano |
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| Rok vydání: | 2007 |
| Předmět: |
Proteases
Bordetella pertussis Bacterial Toxins Molecular Sequence Data Buffers Sodium Chloride Pertussis toxin Biochemistry Mass Spectrometry Microbiology Substrate Specificity Amino Acid Sequence Tromethamine Fluorescent Dyes Corynebacterium diphtheriae chemistry.chemical_classification Gel electrophoresis biology Molecular mass Hydrolysis General Medicine Hydrogen-Ion Concentration biology.organism_classification Culture Media Molecular Weight Enzyme chemistry Pertussis Toxin Chromatography Gel Electrophoresis Polyacrylamide Gel Peptides Bacteria Filtration Biotechnology Peptide Hydrolases |
| Zdroj: | Preparative biochemistrybiotechnology. 37(4) |
| ISSN: | 1082-6068 |
| Popis: | Proteases were identified and characterized from the culture supernatant of the C. diphtheriae and B. pertussis bacteria. The proteases were secreted in the media and detected at the end of the exponential growth phase. Activity was detected in some fluorescent substrates, based on selected protein sequences such as insuline beta-chain, bradykinin, and synaptobrevin. The proteases were purified by means of gel filtration chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the purified proteins indicated, for the main secreted proteins, an estimated molecular mass of 30 kDa in C. diphtheriae and 69 kDa in B. pertussis culture media. The proteases were stable and presented enzymatic activity at 37 degrees C. These proteases were not related to the main toxic compounds described in these two bacteria, but could represent good markers for the fermentation process when the enzyme activity was measured with the fluorescent substrates. |
| Databáze: | OpenAIRE |
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