Depsides and their derivatives are a class of polyketides predominantly found in fungal extracts. Herein, a silent nonreducing polyketide synthase (TalsA)-containing gene cluster, which was identified from the Antarctic sponge-derived fungus Talaromyces sp. HDN1820200, was successfully activated through heterologous expression in Aspergillus nidulans. This activation led to the production of two novel depsides, talaronic acid A (1) and B (2), alongside three known compounds (3–5). The further co-expression of TalsA with the decarboxylase (TalsF) demonstrated that it could convert 2 into its decarboxylated derivative 1. The structural elucidation of these compounds was achieved using comprehensive 1D and 2D-NMR spectroscopy, which was complemented by HR-MS analysis. Talaronic acids A and B were firstly reported heterodimers of 3-methylorsellinic acid (3-MOA) and 5-methylorsellinic acid (5-MOA). All isolated compounds (1–5) were tested for their anti-inflammatory potential. Notably, compounds 1 and 2 exhibited anti-inflammatory activity comparable to that of the positive control. These results further enrich the structural class of depside natural products.