Abstract
Proteinases play a crucial role in invasion and pathogenesis of bacteria, especially the extracellular and membrane-bound forms. Analysis of these proteinases demands the isolation by retaining the enzymatic activity. The isolation procedures maintaining the native structure of the enzyme in its soluble form are also of extreme importance. The qualitative analyses of these proteinases are carried out by electrophoresis and zymography. Enzymatic characterization based on the effect of inhibitors and activators on gelatinase activity also can be assessed using this zymography. The membrane-bound proteinases can be isolated in their native and soluble form, still retaining the activity using 6-aminocaproic acid and sodium deoxycholate; the procedure of which is explained in this chapter.
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Acknowledgments
All the demonstrated experiments were carried out by Mr. Sandhanakrishnan Cattavarayane and the work was supported by grant Nos. SR/SO/HS-29/2005 and SR/S0/HS/0108/2012, SERB Division, Department of Science and Technology, New Delhi, India to M.G.M.
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Madanan, M.G., Mechoor, A. (2017). Detection and Characterization of Bacterial Proteinases Using Zymography. In: Wilkesman, J., Kurz, L. (eds) Zymography. Methods in Molecular Biology, vol 1626. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7111-4_9
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DOI: https://doi.org/10.1007/978-1-4939-7111-4_9
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