Optimization of rapid separation conditions of Ralstonia solanacearum using high performance ion-exchange chromatography

doi:10.3724/SP.J.1123.2016.06044

Abstract

Abstract:

Rapid separation of Ralstonia solanacearum with intact cell using high performance ion-exchange chromatography equipped with a UV detector was described. The bacterial growth abilities and cell morphologies in different suspension buffers were compared. The results showed that the bacterial numbers in the balance buffer (0.02 mol/L piperazine-hydrochloric acid), elution buffer (0.02 mol/L piperazine-hydrochloric acid+1 mol/L NaCl) and double distilled water were 6.467×10⁹, 6.267×10⁹ and 6.233×10⁹ cfu/mL, respectively, exhibiting no significant difference. Moreover, the cell morphologies were observed under a transmission electron microscope which showed that the bacterial cells could remain intact. Effects of buffer pH, flow velocity and bacterial cell concentration on the separation efficiency were analyzed. The results showed that the optimum chromatographic separation parameters were as follows: the buffer pH was 8, the flow velocity was 2 mL/min and the bacterial cell concentration was from 1.0×10⁸ cfu/mL to 1.0×10¹⁰ cfu/mL. Under the chromatographic separation conditions, the separation time was shortened and the separation efficiency was improved. Therefore, the chromatographic method developed here may be a promising tool for the rapid separation of Ralstonia solanacearum and other bacteria.

Key words: Ralstonia solanacearum, cell surface properties, high performance ion-exchange chromatography, separation

CLC Number:

O658

ZHENG Xuefang, LIU Bo, ZHU Yujing, CHEN Deju. Optimization of rapid separation conditions of Ralstonia solanacearum using high performance ion-exchange chromatography[J]. Chinese Journal of Chromatography, 2016, 34(11): 1091-1096.

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