Abstract

VARGAS-ARISPURO, Irasema et al. Looking for natural antifungal agent: Sensitive assey to detect fungal endo-1,3-β-glucanase inhibitors from crude plant extracts. Rev. mex. fitopatol [online]. 2017, vol.35, n.1, pp.130-138. ISSN 2007-8080.  https://doi.org/10.18781/r.mex.fit.1606-5.

1,3-β-glucanase is an enzyme involved in fungal cell wall construction, division septum deposition and ascospore wall assembly. Hence, the 1,3-β-glucanase is a target site to develop into new generations of natural antifungal agents. With the aim of seeking antifungal compounds in plant extracts, we implemented a simple and sensitive assay to detect inhibitors of endo-1,3-β-glucanase from crude plant extracts. The assay, especially useful for screen large number of crude plant extracts, is based on the diffusion of glucanase enzyme from wells through agarosa gel containing the enzyme substrate (laminarin). As the substrate is depolymerized, glucanase diffuses outward in a radial manner leaving a circular zone around the well, unstained by dye calcofluor. A linear relationship between the diameter of hydrolyzed zone and pure enzyme concentration, provide the basis for the method. Results of the assay using plant extracts obtained in methanol or dichloromethane are discussing.

Keywords : Natural fungicide; fungal target site; plant extracts; hydrolase.

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