<p>Dataset used in analysis.</p>

Basidiomycetes are recognized for their capacity to produce a diverse range of secondary metabolites, particularly terpenoids and alkaloids, which have significant applications in the pharmaceutical, agricultural, and chemical industries. Among them, Cristataspora (formerly Humphreya) coffeata has attracted attention for its promising biosynthetic potential. Investigating the range of compounds produced by this fungus in controlled submerged culture conditions, rather than in the fruiting body stage, is particularly beneficial, as it enables precise control of environmental and nutritional factors, resulting in more consistent and scalable metabolite production. This study examined the differences in the cultivation of C. coffeata and the synthesis of terpenoids and alkaloids when the growth medium was supplemented with either glucose or lactose as a carbon source. It was observed that when the fungus was cultivated with glucose, the kinetic parameters did not differ from those with lactose. However, the individual pellet size was larger with glucose compared to lactose. Regarding secondary metabolites, in glucose cultures, a differential production of terpenoids and alkaloids was observed in the biomass and culture supernatant at 6 and 14 days, respectively, compared to lactose, as determined by TLC and spectrophotometric quantification. GC-MS analysis revealed that 19 differential compounds were detected in the biomass and 9 in the supernatant when glucose was used as the carbon source. In contrast, with lactose, 14 compounds were differentially produced in the biomass and 7 in the supernatant. These findings underscore the relevance of submerged cultivation for modulating secondary metabolite profiles and the importance of selecting the appropriate carbon source to maximize yields of target compounds.