In this study, the effects of fungal pretreatment and direct bio‐utilization of RSM for iturin A fermentation were comparatively analyzed through screening suitable fungal species, and evaluating the relationships between iturin A production and the composition of solid fermented RSM and liquid hydrolysates. Three main unconventional adverse effects were identified. In conclusion, our study verifies the unconventional adverse effects of fungal pretreatment on iturin A production by B. amyloliquefaciens CX‐20 compared with direct bio‐utilization of RSM.
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