Abstract
Electric fields have been employed to promote macromolecular crystallization for several decades. Although crystals grown in electric fields seem to present higher diffraction quality, these methods are not widespread. For most configurations, electrodes are in direct contact with the protein solution. Here, we propose a configuration that can be easily extended to standard crystallization methods for which the electrodes are not in direct contact with the protein solution. Furthermore, the proposed electrode configuration supplies an external DC electric field. Glucose Isomerase from Streptomyces rubiginosus crystals were grown at room temperature using the microbatch method in the presence of 1, 2, 4, and 6 kV. Several crystallization trials were carried out for reproducibility and statistical analysis purposes. The comparison with crystals grown in the absence of electric fields showed that crystallization in the presence of electric fields increases the size of crystals, while decreasing the number of nucleations. X-ray diffraction analysis of the crystals showed that those grown in the presence of electric fields are of higher crystal quality.
Highlights
The growth of high quality macromolecular crystals has been addressed theoretically and experimentally by multiple studies
Improved crystal quality for macromolecules crystallized in the presence of internal and external electric fields has been reported by several authors
Initial crystallization trials conducted with the electric fields has been reported by several authors
Summary
The growth of high quality macromolecular crystals has been addressed theoretically and experimentally by multiple studies. The nucleation and growth process of macromolecular crystals [1,2]. Any crystallization method aims to drive a solution from an undersaturated condition to a supersaturation state. Crystal growth rates typically respond to concentration changes at the growing interface and respond to a combination of transport phenomena, such as the diffusion of chemical species and interface kinetics [3]. Several crystallization methods aim to control the former, since any inhomogeneities in the growth solution will lead to convective flow and sedimentation. These may be minimized under low gravity conditions [4,5]. Growth kinetics has been mostly controlled through supersaturation and nucleation inductors, for example through seeding [12,13,14]
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