Nickel-Sepharose (Ni-Sepharose) has been being applied as the most common matrix in purifying His-tag proteins based on the affinity interaction between histidine residues and Ni2+ ion. However, Sepharose still comes at high cost for this purification purpose, especially in developing countries as Vietnam. Here, we show for the first time that melanin from ink sacs of squids which is considered as biowaste in the food industry, can be used as a new potential matrix material instead of Sepharose. We utilized either melanin or melanin charged with metal ions as the stationary phase of affinity purification of His-tag proteins. The results showed that a recombinant His-tag protein VP28 in a protein pool was captured by melanin and Ni2+/Fe3+/Zn2+ chelated melanin. Experiments for releasing VP28 were performed only on the melanin and Ni2+-melanin matrices. The result showed that VP28 was quite selectively eluted when applying elution buffer of 250 mM imidazole overnight. The relative efficiency in releasing VP28 of melanin and Ni-melanin matrices roughly compared to Ni-Sepharose were about 38 and 18% respectively. Further optimization of this process may allow higher efficiency in the purification of His-tag proteins.
Melanin, metal ions, matrix of affinity chromatography, Sepharose, His-tag proteins.
. GE healthcare, “Affinity Chromatography,” GE Healthcare. Handbook., 2007.
. M. d'Ischia , K. Wakamatsu, F. Cicoira , E. Di Mauro, J. C. Garcia-Borron, S. Commo, I. Galván, G. Ghanem, K. Kenzo , P. Meredith, A. Pezzella , C. Santato, T. Sarna, J. D. Simon, L. Zecca, F. A Zucca, A. Napolitano, and S. Ito, “Melanins and melanogenesis: From pigment cells to human health and technological applications,” Pigment Cell and Melanoma Research, vol. 28, no. 5. 2015.
. K. H. Kaidbey, P. P. Agin, R. M. Sayre, and A. M. Kligman, “Photoprotection by melanin-a comparison of black and Caucasian skin,” Journal of the American Acedemy of Dermatology, vol. 1, no. 3, 1979.
. M. Chu, W. Hai, Z. Zhang, F. Wo , Q. Wu, Z. Zhang, Y. Shao, D. Zhang, L. Jin, and D. Shi, “Melanin nanoparticles derived from a homology of medicine and food for sentinel lymph node mapping and photothermal in vivo cancer therapy,” Biomaterials, vol. 91, 2016.
. R. C. R. De Goncalves and S. R. Pombeiro-Sponchiado, “Antioxidant activity of the melanin pigment extracted from Aspergillus nidulans,” Biological & Pharnaceutical. Bulletin, vol. 28, no. 6, 2005.
. A. El-Obeid, S. Al-Harbi, N. Al-Jomah, and A. Hassib, “Herbal melanin modulates tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6) and vascular endothelial growth factor (VEGF) production,” Phytomedicine, vol. 13, no. 5, 2006.
. D. C. Montefiori and J. Zhou, “Selective antiviral activity of synthetic soluble l-tyrosine and l-dopa melanins against human immunodeficiency virus in vitro,” Antiviral Research., vol. 15, no. 1, 1991.
. P. T. Ha, N. H. Nam, D. D. Hai, P. Q. Thong, T. T. M. Nguyet, N. X. Phuc, H. T. M. Nhung, L. M. Huong, N. T. Linh, B. Q. Thuc, “Targeted drug delivery nanosystems based on copolymer poly(lactide)-tocopheryl polyethylene glycol succinate for cancer treatment,” Advances in Natural Sciences: Nanoscience and Nanotechnology, vol. 7, no. 1, 2016.
. A. M. Cuong, N. T. L. Na, P. N. Thang, T. N. Diep, L. B. Thuy, N. L. Thanh, and Nguyen Dinh Thang, “Melanin-embedded materials effectively remove hexavalent chromium (CrVI) from aqueous solution,” Environmental Health and Preventive Medicine, vol. 23, no. 1, pp. 1–11, 2018.
. X. Yu, Z. Gu, R. Shao, H. Chen, X. Wu, and W. Xu, “Study on adsorbing chromium(VI) ions in wastewater by aureobacidium pullulans secretion of melanin,” Advanced. Materials Research., vol. 156-157, pp. 1378-1384, 2011.
. J. D. Nosanchuk and A. Casadevall, “Impact of melanin on microbial virulence and clinical resistance to antimicrobial compounds,” Antimicrobial Agents and Chemotherapy, vol. 50, no. 11. 2006.
. L. Hong, Y. Liu, and J. D. Simon, “Binding of Metal Ions to Melanin and Their Effects on the Aerobic Reactivity,” Photochemistry and Photobiology, vol. 80, no. 3, p. 477, 2004.
. L. Hong and J. D. Simon, “Current Understanding of the Binding Sites, Capacity, Affinity, and Biological Significance of Metals in Melanin,” Society, vol. 111, no. 28, pp. 7938-7947, 2007.
. Y. Liu and J. D. Simon, “The effect of preparation procedures on the morphology of melanin from the ink sac of Sepia officinalis,” Pigment Cell Research, vol. 16, no. 1, 2003,
. N. T. L.Na, S. D. Loc, N. L. M. Tri, N. T. B. Loan, H. A. Son, N. L. Toan, H. P. Thu, H. T. M. Nhung, N. L. Thanh, N. T. V. Anh, and N. D. Thang, “Nanomelanin potentially protects the spleen from radiotherapy-associated damage and enhances immunoactivity in tumor-bearing mice,” Materials (Basel)., vol. 12, no. 10, 2019.
. M. Magarelli, P. Passamonti, and C. Renieri, “Purification, characterization and analysis of sepia melanin from commercial sepia ink (Sepia Officinalis),” Revista CES Medicina Veterinaria y Zootecnia, vol. 5, no. 2, 2010.
. T. Kiss and A. Gergely, “Copper(II) and nickel(II) ternary complexes of l-dopa and related compounds,” Journal of Inorgani Biochemistry, vol. 25, no. 4, pp. 247-259, 1985,
. B. Larsson and H. Tjälve, “Studies on the mechanism of drug-binding to melanin,” Biochem. Pharmacol., vol. 28, no. 7, pp. 1181-1187, 1979.
. K. B. Stȩpień and T. Wilczok, “Studies of the mechanism of chloroquine binding to synthetic dopa-melanin,” Biochemical Pharmacology, vol. 31, no. 21, pp. 3359-3365, 1982.