Biosensors and Nanotheranostics
Bionanotechnology, Drug Delivery, Therapeutics | online ISSN 3064-7789
83
Citations
53.8k
Views
60
Articles
Volume 2 Number 1 2023
RESEARCH ARTICLE (Open Access)
Enhanced Lysozyme Crystallization Using Nano-Templates: Effects of Pore Size and Surface Functionalization
Klepetsanis Pavlos 1*
Biosensors and Nanotheranostics 2 (1) 1-6 https://doi.org/10.25163/biosensors.219839
Submitted: 12 May 2023 Revised: 27 June 2023 Published: 02 July 2023
Abstract
Protein crystallization is a pivotal method in bio-separation, utilized across various fields including structural biology and industrial enzyme production. Recent research has explored nano-templates as a promising approach to enhance protein crystallization by serving as effective nucleants. This study investigates the synthesis and application of nano-templates with varying porosities and surface chemistries to optimize lysozyme crystallization. Nano-templates were synthesized via a sol-gel method using nitric acid and 2M HCl, resulting in average pore sizes of ~4 nm and ~8 nm, respectively. These templates were further functionalized with phenyl, chloro, and methyl groups to modulate their surface properties. Characterization techniques such as nitrogen adsorption-desorption, TEM, FTIR, and zeta potential measurements were employed to assess the templates' properties. Lysozyme crystallization experiments demonstrated that nano-templates significantly reduced the induction time, with phenyl-functionalized templates and those with smaller pore sizes proving most effective. The results indicate that nano-templates with enhanced hydrophobicity and optimal surface charge facilitate faster and more efficient protein nucleation. These findings highlight the potential of nano-templates in advancing protein crystallization techniques, suggesting that further optimization could enhance their utility in bio-separation applications and offer a cost-effective solution for protein purification.
Keywords: Protein crystallization, nano-templates, lysozyme, surface functionalization, pore size
References
Arnold, K., et al. (2007). Silanization of silica surfaces. Journal of Colloid and Interface Science, 315(2), 601-606.
Brinker, C. J., & Scherer, G. W. (1990). Sol-gel science: The physics and chemistry of sol-gel processing. Academic Press.
Chayen, N. E., & Saridakis, E. (2008). Protein crystallization: From purified protein to diffraction-quality crystal. Nature Methods, 5(2), 147-153.
Chen, Y., Zhang, W., & Jiang, T. (2011). Effect of nano-templates on protein crystallization. Journal of Molecular Biology, 406(2), 254-263.
Kim, M. J., Lee, S., & Lee, C. H. (2018). The role of surface functional groups in protein crystallization: A comprehensive review. Biochemical and Biophysical Research Communications, 496(1), 123-132.
Kumar, S., et al. (2021). Silanization techniques for nano-templates: Influence on protein crystallization. Journal of Nanoscience and Nanotechnology, 21(4), 2418-2425.
Li, J., & Zhang, X. (2016). Multifunctional nano-templates for improved protein crystallization. Advanced Functional Materials, 26(15), 2487-2496.
Li, X., Zhang, H., & Yang, Y. (2012). Impact of surface modifications on the crystallization of proteins. CrystEngComm, 14(11), 3870-3878.
Liu, H., et al. (2023). Tailoring nano-template properties for enhanced crystallization of therapeutic proteins. Molecular Pharmaceutics, 20(2), 511-520.
Liu, Y., et al. (2014). Control of membrane protein crystallization using nano-templates. Journal of Structural Biology, 185(1), 33-40.
Matsuura, T., Tanaka, M., & Yamaguchi, T. (2009). Surface functionalization of nano-templates for enhanced protein crystallization. Biophysical Journal, 96(3), 1255-1262.
McPherson, A. (1999). Crystallization of biological macromolecules. Cold Spring Harbor Laboratory Press.
Park, S. H., et al. (2020). Surface charge density modulation in nano-templates: Effects on protein crystallization. Journal of Materials Chemistry B, 8(10), 2295-2304.
Saridakis, E., & Chayen, N. E. (2009). The effect of surface chemistry on protein crystallization. Progress in Biophysics and Molecular Biology, 101(3), 280-289.
Sharma, A., et al. (2024). Nano-template design for efficient protein crystallization: A review. Current Opinion in Biotechnology, 77, 102-111.
Smith, A. D., et al. (2015). Stabilization of fragile proteins during crystallization using nano-templates. Journal of Physical Chemistry B, 119(7), 2361-2370.
Tosi, S., Lipfert, J., & Jedrzejczak, R. (2011). Mechanisms of surface effects on protein crystallization. Biophysical Journal, 100(5), 1201-1210.
Wan, Y., & Zhao, D. (2007). On the role of surfactants in the synthesis of mesoporous structures. Journal of the American Chemical Society, 129(5), 1201-1211.
Wang, Y., et al. (2019). Functional group effects on protein crystallization: Insights from nano-template studies. Journal of Crystal Growth, 510, 35-42.
Yang, L., et al. (2022). Optimizing nano-templates for protein crystallization: Advances and challenges. Journal of Nanomaterials, 2022, Article ID 9276438.
Zhang, Y., et al. (2013). Impact of nano-template surface properties on protein crystallization kinetics. Biomaterials Science, 1(6), 628-636.
Zhao, X., et al. (2017). Enhanced protein crystallization with multifunctional nano-templates. Advanced Materials, 29(46), 1703619
2
Save
Save
0
Citation
Citation
886
View
View
0
Share
Share