Department of Chemistry and Biochemistry, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0358, USA.
Marine Biology Research Division, Scripps Institution of Oceanography, University of California, San Diego, 9500 Gilman Drive La Jolla, California, 92093-0202, USA.
Chembiochem. 2022 May 4;23(9):e202200021. doi: 10.1002/cbic.202200021. Epub 2022 Mar 23.
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.
虽然黑色素是生物体系中最普遍的聚合物之一，但除人类以外，我们对其分子结构、生物合成和物理特性的了解仅限于少数几种生物体。这在一定程度上是由于难以分离出纯黑色素。虽然有一些分离方法，但它们通常涉及使用强酸/强碱条件的苛刻处理，再结合高温，这可能导致聚合物主链降解。成功的分离方法必须能够有选择性地、完全地降解非黑色素生物聚合物，同时去除与黑色素主链不相关的小分子代谢物。在这里，我们展示了荧光探针引导的化学酶处理在纯化和分离天然哺乳动物黑色素方面的应用，而不会显著诱导化学降解。这种多步纯化跟踪方法能够从哺乳动物组织中定量分离出纯黑色素，用于光谱特征分析。
