Characterization of the activity of the spore cortex lytic enzyme CwlJ1
| Autor: | Xia Wu, Ravi S. Kane, Ruchir V. Mundra, Elena E. Paskaleva, Navdeep Grover, Martin A. Page, Jonathan S. Dordick |
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| Rok vydání: | 2015 |
| Předmět: |
chemistry.chemical_classification
Glycan biology fungi Bioengineering biology.organism_classification medicine.disease_cause Applied Microbiology and Biotechnology Bacillus anthracis Amidase Spore Enzyme Biochemistry chemistry Spore germination medicine biology.protein Escherichia coli Bacteria Biotechnology |
| Zdroj: | Biotechnology and Bioengineering. 112:1365-1375 |
| ISSN: | 0006-3592 |
| Popis: | The germination enzyme CwlJ1 plays an important role in degrading the cortex during the germination of Bacillus anthracis spores. However, the specific function and catalytic activity of CwlJ1 remain elusive. Here we report for the first time a detailed in vitro mechanistic study of CwlJ1 expressed in Escherichia coli and its activity against the spore cortical fragments of B. anthracis when added exogenously. CwlJ1 was active on both decoated spores and spore cortical fragments. Through liquid chromatography-mass spectrometry analysis of the digested cortical fragments, we determined that CwlJ1 was a thermostable N-acetylmuramoyl-L-alanine amidase. CwlJ1 mainly recognized large segments of glycan chains in the cortex instead of the minimal structural unit tetrasaccharide, with specificity for muramic acid-δ-lactam-containing glycan chains and preference for the tetrapeptide side chain. Unlike most amidases, CwlJ1 did not appear to contain a divalent cation, as it retained its activity in the presence of EDTA. This study shines some light on the mechanism of spore germination, and provides increased insight into the development of sporicidal enzyme systems for decontamination of B. anthracis and other related bacteria. |
| Databáze: | OpenAIRE |
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