**4. Conclusions**

Our study reported the draft genome of *A. resinae* and characterized its melanin synthesis mechanisms by combining bioinformatics and biochemical approaches. The genome size of *A. resinae* was relatively small than the average genome size of Ascomycetes. Moreover, only 14 BGCs were identified in the genome assembly, which contrasted with the higher BGC abundance of common ascomycetes. Genes encoding a specific T1-PKS and 4-HNR are predicted to produce intermediate metabolites of DHN-melanin biosynthesis but not proceed to DHN-melanin. These findings were further supported by the detection of increased flaviolin concentrations in mycelia and almost unchanged morphologies of the culture grown with tricyclazole. In the melanin formation in culture filtrates, it was observed that laccase-like activity and nitrogen sources are key determinants. Additionally, melanin formation in culture fluid was proportional to the laccase-like activity in the fluid. Melanin synthesis using L-DOPA and the culture filtrate exhibiting laccase-like activity were observed. Therefore, future studies should focus on enhancing laccase-like activity in culture filtrates to optimize industrial-scale melanin production using *A. resinae*.

**Supplementary Materials:** The following materials are available online at https://www.mdpi.com/ article/10.3390/jof7040289/s1, Table S1: Assessment of genome completeness using the BUSCO software. Table S2: Comparisons of genome features between *A. resinae* KUC3009 and its close relatives belonging to the family Myxotrichaeae. Table S3: Putative tyrosinase genes found in the *A. resinae* genome. Table S4: Putative multi-copper oxidase genes found in the *A. resinae* genome.

**Author Contributions:** Conceptualization, J.-J.O.; methodology, J.-J.O., Y.J.K., J.Y.K., S.L.K., C.L., M.- E.L., J.W.K.; formal analysis and investigation, J.-J.O., Y.J.K., J.Y.K.; data curation, J.-J.O., Y.J.K., J.W.K.; writing—original draft preparation, J.-J.O.; writing—review and editing. J.-J.O., Y.J.K., J.Y.K., G.-H.K.; supervision, G.-H.K. All authors have read and agreed to the published version of the manuscript.

**Funding:** This study was supported by a Korea University Research Grant.

**Institutional Review Board Statement:** Not applicable.

**Informed Consent Statement:** Not applicable.

**Data Availability Statement:** Not applicable.

**Conflicts of Interest:** There are no conflict to declare.
