Chemoenzymatic Isolation and Characterization of High Purity Mammalian Melanin. (23rd March 2022)

• Record Type: Journal Article
• Authors: Kalaj, Brianna N.; Ni, Qing Zhe; La Clair, James J.; Deheyn, Dimitri D.; Burkart, Michael D.
• Is Part Of: Chembiochem. Volume 23:Number 9(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-03-23
• Subjects: biopolymers -- chemoenzymatic methods -- enzymes -- fluorescent labeling -- melanin; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.202200021
• Languages: English
• ISSNs: 1439-4227

Abstract

Abstract: Although melanin is one of the most ubiquitous polymers in living systems, our understanding of its molecular structure, biosynthesis and biophysical properties has been limited to only a small number of organisms other than humans. This is in part due to the difficulty associated with isolating pure melanin. While purification methods exist, they typically involve harsh treatments with strong acid/base conditions combined with elevated temperatures that can lead to the polymer backbone degradation. To be successful, a viable isolation method must deliver a selective, yet complete degradation of non‐melanin biopolymers as well as remove small molecule metabolites that are not integrative to the melanin backbone. Here, we demonstrate the use of chemoenzymatic processing guided by fluorescent probes for the purification and isolation of native mammalian melanin without significant induction of chemical degradation. This multi‐step purification‐tracking methodology enables quantitative isolation of pure melanin from mammalian tissue for spectroscopic characterization. Abstract : A quantitative, chemoenzymatic process was developed to isolate eumelanin from mammalian tissues using a fluorescent and electron microscopic screen for guidance. This multi‐step process enables quantitative isolation high purity melanin from mammalian tissue for structural studies as demonstrated by solid‐state NMR analyses.