Supplementary Material for World Mycotoxin Journal: <b>Probiotic and mycotoxin reduction potentials of heat-tolerant </b><b><i>Pichia kudriavzevii</i></b><b> strains from maize- and sorghum-pap</b>

Pap, a gruel prepared by treatment of <i>ogi</i> with boiling water, is widely consumed across West Africa and in the diaspora. It is, however, unknown whether probiotic strains with multi-functional properties can survive the preparation process of pap from <i>ogi</i>. Thus, this study aimed to select the surviving yeasts during the preparation of pap by heat treatment of maize- and sorghum-based <i>ogi</i>, and to screen the yeasts for their probiotic and multi-functional attributes including <i>in vitro</i> mycotoxin reduction potential. Four <i>Pichia kudriavzevii</i> strains out of 16 yeast isolates, including <i>Kurtzmaniella</i><i> natalensis</i> and <i>Kurtzmaniella</i><i> quercitrusa</i>, which survived the heat treatment of <i>ogi</i> to form pap, passed the probiotics selection criteria. These <i>P. kudriavzevii</i> strains were non-haemolytic, utilized bile salt efficiently, and survived 0.3% and 0.5% bile salt concentrations, pH 2.5, gastric and intestinal conditions at 100-104% and 99-104%, 92-99%, 88-92% and 102-121%, respectively. The strains also exhibited up to 97% potential for gut colonization through adherence to hydrocarbons, auto-aggregation activities at 98-100% after 24 h, and co-aggregation of notable foodborne pathogens. In addition, the strains were tolerant to 50 °C incubation for 12 h. The strains’ intact cells and cell-free extracts revealed strong potentials to scavenge and reduce free radicals. LC-MS/MS analysis confirmed that the strains simultaneously reduced the levels of 31 mycotoxins in liquid medium in a 48-h experiment by 1-87%, which is tentatively explained through the efficient adsorption to the cell wall and subsequent metabolization. While reductions of 6-82%, 39-41% and 8-51% were observed for enniatins, sterigmatocystin and zearalenone, respectively, after incubation period of 1 h, all the other investigated mycotoxins were reduced by 1-96% at the 3-h timepoint. Probiotic<i> P. kudriavzevii</i> strains with multifunctional attributes are transferable to pap and other mild heat-treated products for consumer benefits.