Abstract
Melanin is one of the essential compounds in the pigments of molluscan shells. However, the effects of melanin on color variations in molluscs are largely unknown. Our previous study suggests that Yesso scallop Patinopecten yessoensis might contain melanin pigment in the dark brown shell. We therefore isolated melanin from the pigmented shells using hydrochloric acid method, and characterized the types of melanin pigments by spectrophotometry. The purified melanin, which was verified by spectrophotometry scanning and HPLC analysis, showed the typical characteristics of melanin absorption spectra and HPLC chromatograms. The contents of pheomelanin and eumelanin in pigmented shells, which were determined by the linear standard curve of melanin at 405 nm and 350 nm absorbance, were 48.23 ± 1.350 and 157.65 ± 5.905 mg, respectively. The present results indicate that the brown-pigmented shells of scallops comprise approximately 76.6% of eumelanin and 23.4% of pheomelanin, which supports the presence of eumelanin-rich pigment in scallop shells. Therefore, the combination of hydrochloric acid extraction and spectrophotometric quantification is a rapid and efficient method to isolate and quantify melanin in shells. This will facilitate the melanin studies related to shell color polymorphism and the selective breeding of bivalves with different shell colors.
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Arnaud, J. C., and Bore, P., 1981. Isolation of melanin pigments from human hair. Journal of the Society of Cosmetic Chemists, 32: 137–152.
Bai, Z., Zheng, H., Lin, J., Wang, G., and Li, J., 2013. Comparative analysis of the transcriptome in tissues secreting purple and white nacre in the pearl mussel Hyriopsis cumingii. PLoS One, 8: e53617.
Bell, A. A., and Wheeler, M. H., 1986. Biosynthesis and functions of fungal melanins. Annual Review of Phytopathology, 24: 411–451.
Brake, J., Evans, F., and Langdon, C., 2004. Evidence for genetic control of pigmentation of shell and mantle edge in selected families of Pacific oysters, Crassostrea gigas. Aquaculture, 229: 89–98.
Cockell, C. S., and Knowland, J., 1999. Ultraviolet radiation screening compounds. Biological Reviews of the Cambridge Philosophical Society, 74: 311–345.
Comfort, A., 1951. The pigmentation of molluscan shells. Biological Reviews, 26: 285–301.
Del Bino, S., Ito, S., Sok, J., Nakanishi, Y., Bastien, P., Wakamatsu, K., and Bernerd, F., 2015. Chemical analysis of constitutive pigmentation of human epidermis reveals rather constant eumelanin to pheomelanin ratio. Pigment Cell & Melanoma Research, 28: 707–717.
Ding, J., Zhao, L., Chang, Y., Zhao, W., Du, Z., and Hao, Z., 2015. Transcriptome sequencing and characterization of Japanese scallop Patinopecten yessoensis from different shell color lines. PLoS One, 10: e0116406.
d’Ischia, M., Wakamatsu, K., Napolitano, A., Briganti, S., Garcia-Borron, J. C., Kovacs, D., Meredith, P., Pezzella, A., Picardo, M., Sarna, T., Simon, J. D., and Ito, S., 2013. Melanins and melanogenesis: Methods, standards, protocols. Pigment Cell & Melanoma Research, 26: 616–633.
Evans, S., Camara, M. D., and Langdon, C. J., 2009. Heritability of shell pigmentation in the Pacific oyster, Crassostrea gigas. Aquaculture, 286: 211–216.
Ge, J., Li, Q., Yu, H., and Kong, L., 2015a. Identification of single-locus PCR-based markers linked to shell background color in the Pacific oyster (Crassostrea gigas). Marine Biotechnology, 17: 655–662.
Ge, J., Li, Q., Yu, H., and Kong, L., 2015b. Mendelian inheritance of golden shell color in the Pacific oyster Crassostrea gigas. Aquaculture, 441: 21–24.
Guan, Y., and He, M., 2009. Progress on shell coloration polymorphism in seashells. Marine Science Bulletin, 1: 018.
Ito, S., and Fujita, K., 1985. Microanalysis of eumelanin and pheomelanin in hair and melanomas by chemical degradation and liquid chromatography. Analytical Biochemistry, 144: 527–536.
Ito, S., and Wakamatsu, K., 2003. Quantitative analysis of eumelanin and pheomelanin in humans, mice, and other animals: A comparative review. Pigment Cell Research, 16: 523–531.
Ito, S., Nakanishi, Y., Valenzuela, R. K., Brilliant, M. H., Kolbe, L., and Wakamatsu, K., 2011. Usefulness of alkaline hydrogen peroxide oxidation to analyze eumelanin and pheomelanin in various tissue samples: Application to chemical analysis of human hair melanins. Pigment Cell & Melanoma Research, 24: 605–613.
Ito, S., Wakamatsu, K., and Ozeki, H., 1993. Spectrophotometric assay of eumelanin in tissue samples. Analytical Biochemistry, 215: 273–277.
Kumar, R., and Joy, K. P., 2015. Melanins as biomarkers of ovarian follicular atresia in the catfish Heteropneustes fossilis: Biochemical and histochemical characterization, seasonal variation and hormone effects. Fish Physiology and Biochemistry, 41: 761–772.
Liu, Y., He, M., and Wu, L., 2009. Research on the extraction of the melanoma from wugu chicken with the method of acid formulation. Journal of Anhui Agricultural Sciences, 31: 072.
Liu, Y., Kempf, V. R., Brian Nofsinger, J., Weinert, E. E., Rudnicki, M., Wakamatsu, K., Ito, S., and Simon, J. D., 2003. Comparison of the structural and physical properties of human hair eumelanin following enzymatic or acid/base extraction. Pigment Cell Research, 16: 355–365.
Lu, Y., Ye, M., Song, S., Li, L., Shaikh, F., and Li, J., 2014. Isolation, purification, and anti-aging activity of melanin from Lachnum singerianum. Applied Biochemistry and Biotechnology, 174: 762–771.
Lukiewicz, S., 1972. The biological role of melanin. I. New concepts and methodological approaches. Folia Histochemica Cytochemica, 10: 93–108.
Magarelli, M., Passamonti, P., and Renieri, C., 2010. Purification, characterization and analysis of sepia melanin from commercial sepia ink (Sepia officinalis). Revista CES Medicina Veterinariay Zootecnia, 5: 18–28.
Mbonyiryivuze, A., Nuru, Z. Y., Ngom, B. D., Mwakikunga, B., Dhlamini, S. M., Park, E., and Maaza, M., 2015. Morphological and chemical composition characterization of commercial Sepia melanin. American Journal of Nanomaterials, 3: 22–27.
Nagai, K., Yano, M., Morimoto, K., and Miyamoto, H., 2007. Tyrosinase localization in mollusc shells. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 146: 207–214.
Oancea, E., Vriens, J., Brauchi, S., Jun, J., Splawski, I., and Clapham, D. E., 2009. TRPM1 forms ion channels associated with melanin content in melanocytes. Science Signaling, 2: ra21.
Ozeki, H., Ito, S., Wakamatsu, K., and Thody, A. J., 1996. Spectrophotometric characterization of eumelanin and pheomelanin in hair. Pigment Cell Research, 9: 265–270.
Prota, G., 1995. The chemistry of melanins and melanogenesis. In: Fortschritte der Chemie organischer Naturstoffe/Progress in the Chemistry of Organic Natural Products. Springer Vienna, 93–148.
Selvakumar, P., Rajasekar, S., Periasamy, K., and Raaman, N., 2008. Isolation and characterization of melanin pigment from Pleurotus cystidiosus (telomorph of Antromycopsis macrocarpa). World Journal of Microbiology and Biotechnology, 24: 2125–2131.
Slominski, A., Tobin, D. J., Shibahara, S., and Wortsman, J., 2004. Melanin pigmentation in mammalian skin and its hormonal regulation. Physiological Reviews, 84: 1155–1228.
Sun, X., Yang, A., Wu, B., Zhou, L., and Liu, Z., 2015. Characterization of the mantle transcriptome of Yesso scallop (Patinopecten yessoensis): Identification of genes potentially involved in biomineralization and pigmentation. PLoS One, 10: e0122967.
Taylor, S. C., 2002. Skin of color: Biology, structure, function, and implications for dermatologic disease. Journal of the American Academy of Dermatology, 46: S41–S62.
Wakamatsu, K., and Ito, S., 2002. Advanced chemical methods in melanin determination. Pigment Cell Research, 15: 174–183.
Wakamatsu, K., Fujikawa, K., Zucca, F. A., Zecca, L., and Ito, S., 2003. The structure of neuromelanin as studied by chemical degradative methods. Journal of Neurochemistry, 86: 1015–1023.
Wang, H., Pan, Y., Tang, X., and Huang, Z., 2006. Isolation and characterization of melanin from Osmanthus fragrans’ seeds. LWT–Food Science and Technology, 39: 496–502.
Xu, D. Q., and Zhou, J. J., 2013. Research on the extraction technology of the melanin from black feather silky fowl. Food and Fermentation Industries, 3: 046.
Yue, X., Nie, Q., Xiao, G., and Liu, B., 2015. Transcriptome analysis of shell color-related genes in the clam Meretrix meretrix. Marine Biotechnology, 17: 364–374.
Zhang, G., Fang, X., Guo, X, Li, L., Luo, R., Xu, F., Yang, P., Zhang, L., Wang, X., Qi, H., Xiong, Z., Que, H., Xie, Y., Holland, P. W., Paps, J., Zhu, Y., Wu, F., Chen, Y., Wang, J., Peng, C., Meng, J., Yang, L., Liu, J., Wen, B., Zhang, N., Huang, Z., Zhu, Q., Feng, Y., Mount, A., Hedgecock, D., Xu, Z., Liu, Y., Domazet-Lošo, T., Du, Y., Sun, X., Zhang, S., Liu, B., Cheng, P., Jiang, X., Li, J., Fan, D., Wang, W., Fu, W., Wang, T., Wang, B., Zhang, J., Peng, Z., Li, Y., Li, N., Wang, J., Chen, M., He, Y., Tan, F., Song, X., Zheng, Q., Huang, R., Yang, H., Du, X., Chen, L., Yang, M., Gaffney, P. M., Wang, S., Luo, L., She, Z., Ming, Y., Huang, W., Zhang, S., Huang, B., Zhang, Y., Qu, T., Ni, P., Miao, G., Wang, J., Wang, Q., Steinberg, C. E., Wang, H., Li, N., Qian, L., Zhang, G., Li, Y., Yang, H., Liu, X., Wang, J., Yin, Y., and Wang, J., 2012. The oyster genome reveals stress adaptation and complexity of shell formation. Nature, 490: 49–54.
Zheng, H., Zhang, T., Sun, Z., Liu, W., and Liu, H., 2013. Inheritance of shell colours in the noble scallop Chlamys nobilis (Bivalve: Pectinidae). Aquaculture Research, 44: 1229–1235.
Acknowledgements
This work was supported by the grants from the Basic Scientific Research Fund of YSFRI (No. 2060302201516054), the National Natural Science Foundation of China (No. 31602153), and the Natural Science Foundation of Shandong Province (No. ZR2016CQ32).
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Sun, X., Wu, B., Zhou, L. et al. Isolation and characterization of melanin pigment from yesso scallop Patinopecten yessoensis . J. Ocean Univ. China 16, 279–284 (2017). https://doi.org/10.1007/s11802-017-3162-6
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DOI: https://doi.org/10.1007/s11802-017-3162-6