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Objective  The genome of  Vibrio cholerae  has three paralog genes encoding for distinct pyruvate kinases. We were interested in elucidating whether they were expressed, and contributed to the pyruvate kinase activity of  V. cholerae. Vc IPK and  Vc IIPK were transformed and expressed in BL21-CodonPlus(DE3)-RIL strain, whereas  Vc IIIPK could not be transformed. Those studied did contribute to the pyruvate kinase activity of the bacteria. Therefore, our aim was to find an efficient transformation and commonly used over-expression heterologous system for  Vc IIIPK and develop its purification protocol.    Results   vcIpk ,  vcIIpk  and  vcIIIpk  genes were transformed in six different BL21 expression strains. No transformants were obtained for the  vcIIIpk  gene using BL21(DE3), BL21(DE3)pLysS and BL21(DE3)CodonPlus-RIL strains. Reduced rates of cell growth were observed for BL21-Gold(DE3)pLysS and Origami B(DE3)pLysS. High efficiency of transformation was obtained for BL21-AI. Using this strain,  Vc IIIPK was purified but proved to be unstable during its purification and storage. Therefore, the transformation of  vcIIIpk  gene resulted in a toxic, mildly toxic or nontoxic product for these BL21 strains. Despite  Vc IIPK and  Vc IIIPK being phylogenetically related, the preservation of the proteins is drastically different; whereas one is preserved during purification and storage, the other is auto-proteolyzed completely in less than a week.    Electronic supplementary material  The online version of this article (10.1186/s13104-018-3651-8) contains supplementary material, which is available to authorized users.

Exploring the differences between the three pyruvate kinase isozymes from Vibrio cholerae in a heterologous expression system