2019 Fiscal Year Annual Research Report
A multidisciplinary research program to enable the discovery of inhibitors of colanic acid excretion in bacteria
| Project/Area Number |
18H02109
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| Research Institution | Kagawa University |
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Principal Investigator |
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Keywords | antibacterial / single-molecule |
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| Outline of Annual Research Achievements |
The successful cloning of genes of the colanic acid transporter WzaCA into typical bacterial vectors has enabled the expression of WzaCA. Protein expression conditions have been optimised. Single-molecule screening of blockers is ongoing and promising results have been obtained. Chemical strategies to modify potential blockers are being developed. On the other hand, the proposed microscopic system has been partially set up and will be completed in the next fiscal year after purchasing the remaining parts. Initial cellular imaging is ongoing.
Publication of the following paper has been delayed to July 2020.
Lingbing Kong, Revelation of function and inhibition of Wza through single channel studies, Methods Mol. Biol. 2020, in press.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
Due to the reduction of research grant, the essential camera and software for the microscopic system were not purchased in the past fiscal year. These parts will be purchased in the next fiscal year. As a result, a significant part of the Cellular imaging and immunological evaluations of the blockers will be postponed to the next fiscal year.
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| Strategy for Future Research Activity |
Single-molecule screening of blockers will continue to be performed with wild-type and mutant WzaCA protein pores that are expressed and purified under optimised conditions. Once a desired blocker is found, chemical synthesis of stronger blockers derived from this blocker will be carried out based on the chemistry that has been developed in the last fiscal year. Effects of the blockers on colanic acid production in live pathogenic bacteria will continue to be evaluated. Construction of the fluorescence microscope will be completed this fiscal year so that imaging of bacterial cells in the presence and absence of blockers will be performed. Simultaneous electrical and optical recordings of wild-type and mutant WzaCA protein pores interacting with the blockers will be pursued.
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Research Products
(1 results)
