Universidad de Granada (UGR), Departamento de Química Orgánica, Unidad de Excelencia Química Aplicada a Biomedicina y Medioambiente (UEQ), C. U. Fuentenueva, Avda. Severo Ochoa s/n, E-18071 Granada, Spain; Universidad de Granada (UGR), Departamento de Física Aplicada, C. U. Fuentenueva, Avda. Severo Ochoa s/n, E-18071 Granada, Spain; Laboratorio de Estudios Cristalográficos, Instituto Andaluz de Ciencias de la Tierra (Consejo Superior de Investigaciones Científicas-UGR), UEQ, Avenida de las Palmeras 4, 18100 Armilla, Granada, Spain.
Universidad de Granada (UGR), Departamento de Estadística e Investigación Operativa, Spain; Departamento de Mecánica de Estructuras e Ingeniería Hidráulica, Ultrasonics Lab TEP-959, Universidad de Granada, Spain; Unidad de Excelencia Modeling Nature MNAT, Universidad de Granada, Spain; Instituto de Investigación Biosanitaria Ibs, GRANADA, Granada, Spain.
Ultrason Sonochem. 2022 Aug;88:106096. doi: 10.1016/j.ultsonch.2022.106096. Epub 2022 Jul 18.
Sonocrystallization implies the application of ultrasound radiation to control the nucleation and crystal growth depending on the actuation time and intensity. Its application allows to induce nucleation at lower supersaturations than required under standard conditions. Although extended in inorganic and organic crystallization, it has been scarcely explored in protein crystallization. Now, that industrial protein crystallization is gaining momentum, the interest on new ways to control protein nucleation and crystal growth is advancing. In this work we present the development of a novel ultrasound bioreactor to study its influence on protein crystallization in agarose gel. Gel media minimize convention currents and sedimentation, favoring a more homogeneous and stable conditions to study the effect of an externally generated low energy ultrasonic irradiation on protein crystallization avoiding other undesired effects such as temperature increase, introduction of surfaces which induce nucleation, destructive cavitation phenomena, etc. In-depth statistical analysis of the results has shown that the impact of ultrasound in gel media on crystal size populations are statistically significant and reproducible.
超声结晶意味着应用超声辐射来控制成核和晶体生长,具体取决于作用时间和强度。其应用可以在比标准条件下更低的过饱和度下诱导成核。虽然在无机和有机结晶中得到了广泛应用,但在蛋白质结晶中却很少被探索。现在,工业蛋白质结晶正在兴起,人们对控制蛋白质成核和晶体生长的新方法的兴趣也在不断提高。在这项工作中,我们开发了一种新型超声生物反应器,用于研究其对琼脂糖凝胶中蛋白质结晶的影响。凝胶介质最大限度地减少了常规电流和沉淀,有利于更均匀和稳定的条件,以研究外部产生的低能量超声辐射对蛋白质结晶的影响,避免了其他不良影响,如温度升高、引入诱导成核的表面、破坏性空化现象等。对结果的深入统计分析表明,超声在凝胶介质中对晶体尺寸分布的影响在统计学上是显著的,且具有可重复性。