engleski

Disruption of Aspergillus flavus cells: a bead mill homogenization method

The most important mycotoxigenic fungus involved in pre- and post-contamination of crops is Aspergillus flavus which causes great health and economic loses worldwide due to production of the most potent natural hepatocarcinogen – aflatoxin B1. Contamination with this secondary metabolite is getting even worst by global climate changes and other abiotic stressors present in environment. Accordingly, researches with the aim of synthesis or identifying the antiaflatoxigenic and antifungal compounds are of interest. For such efforts realization, use and manipulation with intracellular content of A. flavus cells is necessary. The aim of this study was to apply Omni® Bead Ruptor 12 Homogenizer on disintegration of A. flavus cells, to find optimal parameters of homogenization and prepare biologically active extracts which can be used for determination of possible strategies for control of contamination with aflatoxins. Results of study showed that bead mill homogenizer Omni® Bead Ruptor 12 Homogenizer can be applied for disintegration of A. flavus mycelia and preparation of enzymatically active cell-free extracts. The homogenization mixture in 2 mL homogenization tubes should contain 100 mg of fresh wet mycelia, 1 g of precooled acid washed glass beads of 0.5 mm in diameter and 1 mL of ice- cold buffer. Such mixture should be homogenized at speed of 6 m/s during 120 s, in six cycles of 20 s with cooling of samples in ice-bath between cycles.

bead mill ; disintegration ; Aspergillus flavus ; catalase ; proteins ; aflatoxins

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