Nova Biologica Reperta

Optimization of S-layer protein purification from Deinococcus radiodurans strain R1

Authors

Mahdi Alijanianzadeh 1
Alireza Jalalvand 2
Rasoul Khalilzadeh 3
Maryam Abdolirad 3

1 Department of Cell and Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Karaj, Iran

2 Department of Influenza and Other Respiratory Viruses, Pasteur Institute of Iran, Tehran, Iran

3 Department of Bioscience and Biotechnology, Malek-Ashtar University of Technology, Tehran, Iran

Abstract
S-layer proteins of Deinococcus radiodurans are the best self-assemble systems among other proteins that have an essential role in the fabrication of nanowires. Therefore, the purification of these proteins is necessary. The purpose of this research was to optimize the purification of s-layer protein from D. radiodurans with the response surface method. The three factors of SDS concentration, incubation time and mass percent in five levels were considered, and 20 runs were designed by Design-Expert software with a central composite method. Each run includes microbe culture, mass cell preparation, microbe incubation in specific SDS concentration and time and mass percent, separation of the bacteria from detergent with a centrifuge at 5000g, sedimentation of s-layer proteins from detergent solution with a centrifuge at 20000g, determination of protein concentration, and protein purity by Bradford and SDS-PAGE methods, respectively. Finally, the data obtained were analyzed. Analysis of the results demonstrated that at the 95% confidence level, the effect of the detergent concentration factor on the purified protein percent was more than other factors. The optimization results of factors are 5.64% SDS concentration, 7.33% mass percent, and 3 hours incubation time. At optimized conditions the protein concentration and purity percent were obtained 0.584 mg/ml and 47.61% respectively.


Keywords

Electrophoresis
detergent
purification
response surface
surface layer protein
Abdolirad, M., Khalilzadeh R. & Alijanianzadeh, M. 2016. Isolation and study of s-layer nanostructure of Deinococcus radiodurans R1. Biomacromolecular Journal 2: 126-134.
Abdolirad, M., khalilzadeh, R. & Alijanianzadeh, M. 2019. Self-assembly of gold nanoparticles using HPI s-layer proteins biotemplate. Biomacromolecular Journal 5: 152-160.
Baumeister W Fau - Karrenberg, F., Karrenberg F Fau - Rachel, R., Rachel R Fau - Engel, A., Engel A Fau - ten Heggeler, B., ten Heggeler B Fau - Saxton, W.O. & Saxton, W.O. 1982. The major cell envelope protein of Micrococcus radiodurans (R1). Structural and chemical characterization. European Journal of Biochemistry 125: 535-544.
Bharat, T.A.M., von Kügelgen, A. & Alva, V. 2021. Molecular logic of prokaryotic surface layer structures. Trends in Microbiology 29: 405-415.
Bhuyan, A. K. 2010. On the mechanism of SDS-induced protein denaturation. Biopolymers 93: 186-199.
Cui, H.-F., Ye, J.-S., Leitmannova Liu, A., & Ti Tien, H. 2006. Chapter 1: Lipid Microvesicles: On the Four Decades of Liposome Research. In A. L. Liu (Ed.), Advances in Planar Lipid Bilayers and Liposomes (Vol. 4, pp. 1-48). Academic Press.
Ebadi Sharaf Abad, B., Khalilzadeh, R., Alijanianzadeh, M., & Abdolirad, M. 2017. Investigation of effective factors on the optimization of surface layer protein production in the Deinococcus radiodurans R1 strain using response surface method. Iranian Jouornal of Medical Microbiology 11: 59-70.
Fagan, R. P., & Fairweather, N. F. 2014. Biogenesis and functions of bacterial S-layers. Nature Reviews Microbiology 12: 211-222.
Farci, D., Aksoyoglu, M.A., Farci, S.F., Bafna, J.A., Bodrenko, I., Ceccarelli, M., & Piano, D. 2020. Structural insights into the main S-layer unit of Deinococcus radiodurans reveal a massive protein complex with porin-like features. J Biol Chem 295: 4224-4236.
Farci, D., Bowler, M.W., Esposito, F., McSweeney, S., Tramontano, E., & Piano, D. 2015. Purification and characterization of (DR_2577 ((SlpA) a major S-layer protein from Deinococcus radiodurans. Frontiers in Microbiology 6: 414-419.
Farci, D., Slavov, C., Tramontano, E., & Piano, D. 2016. The S-layer Protein DR_2577 Binds Deinoxanthin and under Desiccation Conditions Protects against UV-Radiation in Deinococcus radiodurans. Frontiers in Microbiology 7: 155-163.
Habibi, N., Caneva Soumetz, F., Giulianelli, M., Pastorino, L., Herrera, O., Sbrana, F., & Ruggiero, C. 2010. Self-assembly and recrystallization of bacterial S-layer proteins of Bacillus sphaericus and Bacillus thuringiensis on silicone, mica and quartz crystal supports. Annu Int Conf IEEE Eng Med Biol Soc 37: 3742-3749.
Herrmann, J., Li, P.-N., Jabbarpour, F., Chan, A.C., Rajkovic, I., Matsui, T., & Murphy, M. E. 2020. A bacterial surface layer protein exploits multistep crystallization for rapid self-assembly. Proceedings of the National Academy of Sciences 117: 388-394.
Hollmann, L.D. & Miyoshi, A. 2010. S-layer proteins from lactobacilli as vaccine delivery systems. International Journal of Microbiology Research 2: 30-43.
Klotz, C., Goh, Y. J., O’Flaherty, S., & Barrangou, R. 2020. S-layer associated proteins contribute to the adhesive and immunomodulatory properties of Lacto bacillus acidophilus NCFM. BMC Microbiology 20: 248-253.
Li, F., Wang, D., Zhou, J., Men, D., & Zhan, X. E. 2020. Design and biosynthesis of functional protein nanostructures. Science China Life Sciences 63: 1142-1158.
Madhurantakam, C., Howorka, S., & Remaut, H. 2014. S-layer Structure in Bacteria and Archaea. In L.L. Barton, D.A. Bazylinski, & H. Xu (Eds.), Nanomicrobiology: Physiological and Environmental Characteristics (pp. 11-37). Springer New York.
Mark, S. S., Bergkvist M Fau - Yang, X., Yang X Fau - Teixeira, L. M., Teixeira Lm Fau - Bhatnagar, P., Bhatnagar P Fau - Angert, E. R., Angert Er Fau Batt, C.A., & Batt, C.A. 2006. Bionanofabrication of metallic and semiconductor nanoparticle arrays using S-layer protein lattices with different lateral spacings and geometries. Langmuir 22: 3763-3774.
Misra, C.S., Basu, B., & Apte, S.K. 2015. Surface (S)-layer proteins of Deinococcus radiodurans and their utility as vehicles for surface localization of functional proteins. Biochimica et Biophysica Acta 1848: 3181-3187
Müller, D. J., Baumeister W Fau - Engel, A., & Engel, A. 1996. Conformational change of the hexagonally packed intermediate layer of Deinococcus radiodurans monitored by atomic force microscopy. Journal of Bacteriology 178: 3025-3030.
Pum, D., Breitwieser, A., & Sleytr, U.B. 2021. Patterns in Nature S-Layer Lattices of Bacterial and Archaeal Cells. Crystals 11: 869-874.
Pum, D., Messner, P., & Sleytr, U.B. 1991. Role of the S layer in morphogenesis and cell division of the archaebacterium Methanocorpusculum sinense. Journal of bacteriology 173: 6865-6873.
Pum, D., Toca-Herrera, J. L., & Sleytr, U.B. 2013. S-layer protein self-assembly. International Journal of Molecular Sciences 14: 2484-2501.
Rachel, R., Engel, A., & Baumeister, W. 1983. Proteolysis of the major cell envelope protein of Deinococcus radiodurans remains morphologically latent. FEMS Microbiology Letters 17: 115-119.
Rothfuss, H., Lara, J.C., Schmid, A. K., & Lidstrom, M. E. 2006. Involvement of the S-layer proteins Hpi and SlpA in the maintenance of cell envelope integrity in Deinococcus radiodurans R1. Microbiology (Reading) 152: 2779-2787.
Sára, M., & Sleytr, U.B. 1987. Molecular sieving through S layers of Bacillus stearothermophilus strains. Journal of Bacteriology 169: 4098-4102.
Schuster, B. 2018. S-Layer Protein-Based Biosensors. Biosensors 82: 40-47.
Schuster, B., & Sleytr, U.B. 2013. Nanotechnology with S-Layer Proteins. In J. A. Gerrard (Ed.), Protein Nanotechnology: Protocols, Instrumentation, and Applications, Second Edition (pp. 153-175). Humana Press.
Schuster, B., & Sleytr, U.B. 2014. Biomimetic interfaces based on S-layer proteins, lipid membranes and functional biomolecules. Journal of the Royal Society, Interface 11: 20140232-20140232.
Shukla, S. K., Manobala, T., & Rao, T.S. 2022. The role of S-layer protein (SlpA) in biofilm-formation of Deinococcus. Journal of Applied Microbiology 133: 796-807.
Simonin, P., Lombard, C., Huguet, A., & Kish, A. 2020. Improved Isolation of SlaA and SlaB S-layer proteins in Sulfolobus acidocaldarius. Extremophiles 24: 673-680.
Sleytr, U.B., & Beveridge, T.J. 1999. Bacterial S-layers. Trends in Microbiology 7: 253-260.
Nova Biologica Reperta
Volume 9, Issue 4 - Serial Number 34
Autumn 2022
Pages 267-278

  • Receive Date 08 June 2026
  • Publish Date 08 June 2026

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