Regulation of Cunninghamella spp. biofilm growth by tryptophol and tyrosol

Fungi belonging to the genus Cunninghamella are often used as microbial models of mammalian metabolism owing to their ability to transform a range of xenobiotic compounds. Furthermore, under specific growth conditions species such as Cunninghamellaelegans and Cunninghamellaechinulata grow as biofilms enabling a convenient semi-continuous production of valuable drug metabolites. However, the molecular mechanism of biofilm regulation is not understood, thus controlling biofilm thickness limits the productive applications of it. In this paper we describe the identification of two molecules, tyrosol and tryptophol, that were identified in C. blakesleeana cultures, but not in C. elegans and C. echinulata. The molecules are known quorum sensing molecules (QSMs) in yeast and their potential role in Cunninghamella biofilm regulation was explored. Both were present in higher concentrations in C. blakesleeana planktonic cultures compared with biofilms; they inhibited the growth of the fungus on agar plates and selectively inhibited biofilm growth in liquid cultures. The molecules had a comparatively minor impact on the biofilm growth of C. elegans and C. echinulata and on the growth of these fungi on agar plates. Finally, when exogenous tyrosol or tryptophol was added to previously grown C. blakesleeana biofilm, detachment was visible and new additional planktonic culture was measured, confirming that these molecules specifically regulate biofilm growth in this fungus.