A fast protein liquid chromatography method for purification of myoglobin from different species

Year 2023, Volume: 8 Issue: 2, 95 - 103, 31.08.2023

Bahram Sarkaratı Nursel Söylemez Milli Omer Eren

https://doi.org/10.24880/maeuvfd.1230538

Abstract

The aim of this study is to describe a fast method for the purification of high-purity myoglobin for Mass Spectroscopy analyses and to use it as standard-grade material. A three-step Fast Protein Liquid Chromatography (FPLC) method was used to produce high-purity myoglobin. SEC 650 gel filtration followed by an Enrich Q anion exchange chromatography was used to produce myoglobin in acceptable purity for most research methods. A second filtration step was carried out by narrow field SEC 70 gel to prepare high-purity myoglobin at standard grade purity and capable of Mass Spectroscopy analyses. At least 90% pure myoglobin was obtained by applying two chromatography steps in the samples of three species, and over 99% pure myoglobin was obtained in standard material quality and suitable for mass spectroscopy when the additional narrow field SEC 70 chromatography step was applied. The proposed method provides higher purity compared to other methods and can be applied in a shorter time. FPLC columns significantly reduce the duration of the chromatography steps. At the same time, the use of solid extraction columns instead of dialysis reduced the long overnight dialysis process to a few minutes.

Keywords

myoglobin, FPLC, protein purification, species identification

Supporting Institution

Bolu Abant Izzet Baysal University

Project Number

2015.09.04.912

Thanks

The authors would like to thank the Scientific Industrial and Technological Application and Research Center (STARC) of Bolu Abant İzzet Baysal University for the utilization of laboratories.

References

• REFERENCES 1. Chaijan, M., Benjakul, S., Visessanguan, W., & Faustman, C. (2007). Characterisation of myoglobin from sardine (Sardinella gibbosa) dark muscle. Food Chemistry, 100(1), 156–164. https://doi.org/10.1016/j.foodchem.2005.09.030
• 2. Chotichayapong, C., Sattayasai, N., Kanzawa, N., Tamiya, T., Tsuchiya, T., & Chanthai, S. (2016). Purification, peptide mapping and spectroscopic characterization of myoglobin from striped snake-head fish (ophicephalus striatus). Oriental Journal of Chemistry, 32(1), 181–194. https://doi.org/10.13005/OJC/320119
• 3. Di Giuseppe, A. M. A., Russo, R., Ragucci, S., Landi, N., Rega, C., Chambery, A., & Di Maro, A. (2018). Myoglobin from common pheasant (Phasianus colchicus L.): Purification and primary structure characterization. Journal of Food Biochemistry, 42(2), 1–9. https://doi.org/10.1111/jfbc.12477
• 4. Di Giuseppe, A. M. A., Russo, L., Russo, R., Ragucci, S., Caso, J. V., Isernia, C., Chambery, A., & Di Maro, A. (2017). Molecular characterization of myoglobin from Sciurus vulgaris meridionalis: Primary structure, kinetics and spectroscopic studies. Biochimica et Biophysica Acta - Proteins and Proteomics, 1865(5), 499–509. https://doi.org/10.1016/j.bbapap.2017.02.011
• 5. Dosi, R., Carusone, A., Chambery, A., Severino, V., Parente, A., & Di Maro, A. (2012). Rapid primary structure determination of myoglobins by a complementary approach based on mass spectrometry and Edman degradation. Food Chemistry, 133(4), 1646–1652. https://doi.org/10.1016/j.foodchem.2012.02.042
• 6. Dosi, R., Di Maro, A., Chambery, A., Colonna, G., Costantini, S., Geraci, G., & Parente, A. (2006). Characterization and kinetics studies of water buffalo (Bubalus bubalis) myoglobin. Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology, 145(2), 230–238. https://doi.org/10.1016/j.cbpb.2006.07.006
• 7. Enoki, Y., Ohga, Y., Ishidate, H., & Morimoto, T. (2008). Primary structure of myoglobins from 31 species of birds. Comparative Biochemistry and Physiology - B Biochemistry and Molecular Biology, 149(1), 11–21. https://doi.org/10.1016/j.cbpb.2007.07.006
• 8. Gatellier, P., Anton, M., & Renerre, M. (1993). A rapid method of oxymyoglobin purification. Meat Science, 33(3), 401–407. https://doi.org/10.1016/0309-1740(93)90011-6
• 9. Giaretta, N., Di Giuseppe, A. M. A., Lippert, M., Parente, A., & Di Maro, A. (2013). Myoglobin as marker in meat adulteration: a UPLC method for determining the presence of pork meat in raw beef burger. Food Chemistry, 141(3), 1814–1820. https://doi.org/10.1016/J.FOODCHEM.2013.04.124
• 10. Gotoh, T., & Shikama, K. (1974). Autoxidation of native oxymyoglobin from bovine heart muscle. Archives of Biochemistry and Biophysics, 163(2), 476–481. https://doi.org/10.1016/0003-9861(74)90504-9
• 11. Joseph, P., Suman, S. P., Li, S., Beach, C. M., & Claus, J. R. (2010). Mass spectrometric characterization and thermostability of turkey myoglobin. LWT - Food Science and Technology, 43(2), 273–278. https://doi.org/10.1016/J.LWT.2009.08.019
• 12. Joseph, P., Suman, S. P., Li, S., Beach, C. M., Steinke, L., & Fontaine, M. (2010). Characterization of bison (Bison bison) myoglobin. Meat Science, 84(1), 71–78. https://doi.org/10.1016/j.meatsci.2009.08.014
• 13. Joseph, P., Suman, S. P., Li, S., Claus, J. R., Fontaine, M., & Steinke, L. (2011). Primary structure of turkey myoglobin. Food Chemistry, 129(1), 175–178. https://doi.org/10.1016/j.foodchem.2011.04.024
• 14. Laemmli, U. K. (1970). Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature, 227(5259), 680–685. https://doi.org/10.1038/227680a0
• 15. Mottola, A., Piredda, R., Catanese, G., Lorusso, L., Ciccarese, G., & Di Pinto, A. (2022). Species authentication of canned mackerel: Challenges in molecular identification and potential drivers of mislabelling. Food Control, 137(January), 108880. https://doi.org/10.1016/j.foodcont.2022.108880
• 16. Nurilmala, M., Ushio, H., Watabe, S., & Ochiai, Y. (2018). A streamlined isolation method and the autoxidation profiles of tuna myoglobin. Journal of Food Science and Technology, 55(5), 1641–1647. https://doi.org/10.1007/s13197-018-3068-3 17. Ordway, G. A., & Garry, D. J. (2004). Myoglobin: An esse ntial hemoprotein in striated muscle. Journal of Experimental Biology, 207(20), 3441–3446. https://doi.org/10.1242/jeb.01172
• 18. Ragucci, S., Russo, R., Landi, N., Valletta, M., Chambery, A., Esposito, S., Raundrup, K., & Di Maro, A. (2019). Muskox myoglobin: purification, characterization and kinetics studies compared with cattle and water buffalo myoglobins. Journal of the Science of Food and Agriculture, 99(14), 6278–6286. https://doi.org/10.1002/jsfa.9901
• 19. Ragucci, S., Woodrow, P., Clemente, A., Russo, R., Valletta, M., Landi, N., Russo, L., Chambery, A., & Di Maro, A. (2022). Myoglobin from Atlantic and Tinker mackerels: Purification, characterization and its possible use as a molecular marker. International Journal of Biological Macromolecules, 214(June), 459–469. https://doi.org/10.1016/j.ijbiomac.2022.06.128
• 20. Renerre, M., Anton, M., & Gatellier, P. (1992). Autoxidation of purified myoglobin from two bovine muscles. Meat Science, 32(3), 331–342. https://doi.org/10.1016/0309-1740(92)90096-M
• 21. Romero-Herrera, A. E., Lehmann, H., Castillo, O., Joysey, K. A., & Friday, A. E. (1976). Myoglobin of the orangutan as a phylogenetic enigma. Nature 1976 261:5556, 261(5556), 162–164. https://doi.org/10.1038/261162a0
• 22. Saud, S., Li, G., Kong, H., Khan, M. I., Qiang, Z., Sun, Y., Liu, W., Ding, C., Xiao, H., Wang, Y., Li, H., & Cao, C. (2019). Identification of chicken meat quality via rapid array isoelectric focusing with extraction of hemoglobin and myoglobin in meat sample. Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, 1128(August), 121790. https://doi.org/10.1016/j.jchromb.2019.121790
• 23. Shikama, K., & Sugawara, Y. (1978). Autoxidation of native oxymyoglobin. Kinetic analysis of the pH profile. European Journal of Biochemistry, 91(2), 407–413. https://doi.org/10.1111/J.1432-1033.1978.TB12693.X
• 24. Suman, S. P., Joseph, P., Li, S., Steinke, L., & Fontaine, M. (2009). Primary structure of goat myoglobin. Meat Science, 82(4), 456–460. https://doi.org/10.1016/j.meatsci.2009.02.017
• 25. Suzuki, T., Sugawara, Y., Satoh, Y., & Shikama, K. (1980). Human oxymyoglobin: isolation and characterization. Journal of Chromatography A, 195(2), 277–280. https://doi.org/10.1016/S0021-9673(00)96820-1
• 26. Tan, B., Sun, B., Sun, N., Li, C., Zhang, J., & Yang, W. (2021). Structure, functional properties and iron bioavailability of Pneumatophorus japonicus myoglobin and its glycosylation products. International Journal of Biological Macromolecules, 173, 524–531. https://doi.org/10.1016/j.ijbiomac.2021.01.138
• 27. Yamazaki, I., Yokota, K. N., & Shikama, K. (1964). Preparation of Crystalline Oxymyoglobin from Horse Heart. Journal of Biological Chemistry, 239(12), 4151–4153. https://doi.org/10.1016/S0021-9258(18)91147-6