The protective role and mechanism of melanin for Aspergillus niger and Aspergillus flavus against chlorine-based disinfectants.

Melanin is a critical component of fungal cell wall which protect fungi from adverse environmental tress. However, the role of melanin for fungi during the disinfection with chlorine-based disinfectants has not been elucidated. The results showed that the inactivation rate constants of Aspergillus niger with chlorine and chlorine dioxide decreased from 0.08 to 2.10 min to 0 after addition of 0.32 mg/L melanin. The results indicated addition of extracted fungal melanin inhibited the inactivation efficiency of chlorine and chlorine dioxide. In contrast, the k of Aspergillus niger after inactivation with monochloramine ranged from 1.50 to 1.78 min after addition of melanin which indicated effect of melanin on the inactivation efficiency of monochloramine was negligible. In addition, the extracted fungal melanin exhibited high reactivity with chlorine and chlorine dioxide but very low reactivity with monochloramine. The different inactivation mechanisms of chlorine-based disinfectants and different reactivity of melanin with chlorine-based disinfectants led to the different protective mechanism of melanin for A. niger and A. flavus spores against disinfection with chlorine-based disinfectants. The chlorine and chlorine dioxide appeared to react with functional groups of melanin in cell wall of spores, so sacrificial reactions between melanin and disinfectants decreased the available disinfectants and limited the diffusion of disinfectants to the reactive site on cell membrane, which led to the decrease of the disinfection efficiency for chlorine and chlorine dioxide. The monochloramine could penetrate into cell and damage DNA without the effect of melanin due to its strong penetration and low reactivity with melanin. Our results systematically demonstrate the protective roles of melanin on the fungal spores against chlorine-based disinfectants and the underlying mechanisms in resisting the environmental stress caused by chlorine-based disinfectants, which provides important implications for the control of fungi, especially for fungi producing melanin.