*Article* **Genomic Analysis and Assessment of Melanin Synthesis in** *Amorphotheca resinae* **KUC3009**

**Jeong-Joo Oh <sup>1</sup> , Young Jun Kim <sup>2</sup> , Jee Young Kim 1, Sun Lul Kwon 1, Changsu Lee 3, Myeong-Eun Lee 4, Jung Woo Kim <sup>5</sup> and Gyu-Hyeok Kim 1,\***


**Abstract:** This study reports the draft genome of *Amorphotheca resinae* KUC30009, a fungal isolate with promising industrial-scale melanin production potential. The mechanisms for melanin or melaninrelated pigment formation of this strain were examined through bioinformatic and biochemical strategies. The 30.11 Mb genome of *A. resinae* contains 9638 predicted genes. Genomic-based discovery analyses identified 14 biosynthetic gene clusters (BGCs) associated with secondary metabolite production. Moreover, genes encoding a specific type 1 polyketide synthase and 4-hydroxynaphthalene reductase were identified and predicted to produce intermediate metabolites of dihydroxy naphthalene (DHN)-melanin biosynthesis pathway, but not 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. Apart from this, the formation of melanin in the culture filtrate appeared to depend on the laccase-like activity of multi-copper oxidases. Simultaneously, concentrations of nitrogen-containing sources decreased when the melanin formed in the media. Interestingly, melanin formation in the culture fluid was proportional to laccase-like activity. Based on these findings, we proposed novel strategies for the enhancement of melanin production in culture filtrates. Therefore, our study established a theoretical and methodological basis for synthesizing pigments from fungal isolates using genomic- and biochemical-based approaches.

**Keywords:** *Amorphotheca resinae*; fungal melanin; bioinformatics; melanin pigments
