Characterization of the Escherichia coli Antifungal Protein PPEBL21
An antifungal protein isolated from Escherichia coli BL21 (PPEBL21) and predicted to be alcohol dehydrogenase (ADH) was subjected to biological characterization. The PPEBL21, indeed, demonstrated propionaldehyde-specific ADH activity. The Km and Vmax of PPEBL21 were found to be 644.8 μM and 1.2 U/mg...
Saved in:
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Wiley
2010-01-01
|
| Series: | International Journal of Microbiology |
| Online Access: | http://dx.doi.org/10.1155/2010/196363 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832554035147702272 |
|---|---|
| author | V. Yadav R. Mandhan M. Kumar J. Gupta G. L. Sharma |
| author_facet | V. Yadav R. Mandhan M. Kumar J. Gupta G. L. Sharma |
| author_sort | V. Yadav |
| collection | DOAJ |
| description | An antifungal protein isolated from Escherichia coli BL21 (PPEBL21) and predicted to be alcohol dehydrogenase (ADH) was subjected to biological characterization. The PPEBL21, indeed, demonstrated propionaldehyde-specific ADH activity. The Km and Vmax of PPEBL21 were found to be 644.8 μM and 1.2 U/mg, respectively. In-gel activity assay also showed that PPEBL21 was a propionaldehyde-specific ADH. The pI of PPEBL21 was observed to be 7.8. PPEBL21 was found to be stable up to a temperature of 40∘C with optimum activity at pH 7.5. The decrease in pH decreased the activity of PPEBL21. These results suggested that PPEBL21 having alcohol dehydrogenase activity and stability at significantly high temperature might be an important lead antifungal molecule. Experiments were performed to identify the possible target of PPEBL21 in the pathogen A. fumigatus. Results revealed that PPEBL21 inhibited completely the expression of a 16 kDa protein in A. fumigatus. The 16 kDa protein of A. fumigatus targeted by PPEBL21 was identified as a hypothetical protein by peptide mass fingerprinting. It is thus hypothesized that a 16 kDa factor is essentially required by A. fumigatus for survival and its impaired synthesis due to treatment with PPEBL21 may lead to the death of pathogen. |
| format | Article |
| id | doaj-art-ae6c352077e44f1cb53ef18457184a35 |
| institution | Kabale University |
| issn | 1687-918X 1687-9198 |
| language | English |
| publishDate | 2010-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | International Journal of Microbiology |
| spelling | doaj-art-ae6c352077e44f1cb53ef18457184a352025-02-03T05:52:39ZengWileyInternational Journal of Microbiology1687-918X1687-91982010-01-01201010.1155/2010/196363196363Characterization of the Escherichia coli Antifungal Protein PPEBL21V. Yadav0R. Mandhan1M. Kumar2J. Gupta3G. L. Sharma4National Institute for Health, Rockville Pike, Bethesda, MD 20892, USADepartment of Biotechnology, Kurukshetra University, Kurukshetra 136119, IndiaInstitute of Genomics and Integrative Biology, Mall Road, Delhi 110007, IndiaResearch Institute of the McGill University Health Centre, Montréal, QC, H3G 1A4, CanadaInstitute of Genomics and Integrative Biology, Mall Road, Delhi 110007, IndiaAn antifungal protein isolated from Escherichia coli BL21 (PPEBL21) and predicted to be alcohol dehydrogenase (ADH) was subjected to biological characterization. The PPEBL21, indeed, demonstrated propionaldehyde-specific ADH activity. The Km and Vmax of PPEBL21 were found to be 644.8 μM and 1.2 U/mg, respectively. In-gel activity assay also showed that PPEBL21 was a propionaldehyde-specific ADH. The pI of PPEBL21 was observed to be 7.8. PPEBL21 was found to be stable up to a temperature of 40∘C with optimum activity at pH 7.5. The decrease in pH decreased the activity of PPEBL21. These results suggested that PPEBL21 having alcohol dehydrogenase activity and stability at significantly high temperature might be an important lead antifungal molecule. Experiments were performed to identify the possible target of PPEBL21 in the pathogen A. fumigatus. Results revealed that PPEBL21 inhibited completely the expression of a 16 kDa protein in A. fumigatus. The 16 kDa protein of A. fumigatus targeted by PPEBL21 was identified as a hypothetical protein by peptide mass fingerprinting. It is thus hypothesized that a 16 kDa factor is essentially required by A. fumigatus for survival and its impaired synthesis due to treatment with PPEBL21 may lead to the death of pathogen.http://dx.doi.org/10.1155/2010/196363 |
| spellingShingle | V. Yadav R. Mandhan M. Kumar J. Gupta G. L. Sharma Characterization of the Escherichia coli Antifungal Protein PPEBL21 International Journal of Microbiology |
| title | Characterization of the Escherichia coli Antifungal Protein PPEBL21 |
| title_full | Characterization of the Escherichia coli Antifungal Protein PPEBL21 |
| title_fullStr | Characterization of the Escherichia coli Antifungal Protein PPEBL21 |
| title_full_unstemmed | Characterization of the Escherichia coli Antifungal Protein PPEBL21 |
| title_short | Characterization of the Escherichia coli Antifungal Protein PPEBL21 |
| title_sort | characterization of the escherichia coli antifungal protein ppebl21 |
| url | http://dx.doi.org/10.1155/2010/196363 |
| work_keys_str_mv | AT vyadav characterizationoftheescherichiacoliantifungalproteinppebl21 AT rmandhan characterizationoftheescherichiacoliantifungalproteinppebl21 AT mkumar characterizationoftheescherichiacoliantifungalproteinppebl21 AT jgupta characterizationoftheescherichiacoliantifungalproteinppebl21 AT glsharma characterizationoftheescherichiacoliantifungalproteinppebl21 |