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软层特性对细菌电穿孔影响的数值研究。

Numerical study of the effect of soft layer properties on bacterial electroporation.

机构信息

Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, United States; Department of Mechanical Engineering, George Mason University, Fairfax, VA 22030, United States.

Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, United States.

出版信息

Bioelectrochemistry. 2018 Oct;123:261-272. doi: 10.1016/j.bioelechem.2017.09.004. Epub 2017 Sep 18.

Abstract

We present a numerical model of electroporation in a gram-positive bacterium, which accounts for the presence of a negatively charged soft polyelectrolyte layer (which may include a periplasmic space, peptidoglycan layer, cilia, flagella, and other surface appendages) surrounding its plasma membrane. We model the ion transport within and outside the soft layer using the soft layer electrokinetics-based Poisson-Nernst-Planck formalism. Additionally, we model the electroporation dynamics on the plasma membrane using the pore nucleation-based electroporation formalism developed by Krassowska and Filev. We find that ion transport within the soft layer (surface conduction), which depends on the relative importance of the soft layer charged group concentration compared to the buffer concentration, significantly alters the transmembrane voltage across the plasma membrane and hence the pore characteristics. Our numerical simulations suggest that surface conduction significantly lowers the pore number in the plasma membrane. This observation is consistent with experimental studies that show that gram-positive bacteria, in general, have lower transformation efficiencies compared to gram-negative bacteria. Our studies highlight a strong dependence of bacterial electroporation on cell envelope properties and buffer conditions, which need to be taken into consideration when designing electroporation protocols.

摘要

我们提出了一种革兰氏阳性菌电穿孔的数值模型,该模型考虑到了带负电荷的软聚电解质层(可能包括周质空间、肽聚糖层、纤毛、鞭毛和其他表面附属物)围绕其质膜的存在。我们使用基于软层电动动力学的泊松-纳恩斯-普朗克公式来模拟软层内外的离子传输。此外,我们使用由 Krassowska 和 Filev 开发的基于孔核理论的电穿孔形式来模拟质膜上的电穿孔动力学。我们发现,软层内的离子传输(表面传导)取决于软层带电基团浓度与缓冲液浓度的相对重要性,这会显著改变质膜的跨膜电压,从而影响孔的特性。我们的数值模拟表明,表面传导会显著降低质膜上的孔数。这一观察结果与实验研究一致,表明革兰氏阳性菌的转化效率一般低于革兰氏阴性菌。我们的研究强调了细菌电穿孔对细胞膜特性和缓冲条件的强烈依赖性,在设计电穿孔方案时需要考虑这些因素。

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