This review reports results from our laboratory on the development of an effective inducible expression system for the homologous/heterologous protein production in cold-adapted bacteria. Recently, we isolated and characterized a regulative genomic region from Pseudoalteromonas haloplanktis TAC125; in particular, a two-component regulatory system was identified. It is involved in the transcriptional regulation of the gene coding for an outer membrane porin (PSHAb0363) that is strongly induced by the presence of l-malate in the growth medium. We used the regulative region comprising the two-component system located upstream the PSHAb0363 gene to construct an inducible expression vector - named pUCRP - under the control of l-malate. Performances of the inducible system were tested using the psychrophilic β-galactosidase from P. haloplanktis TAE79 as model enzyme to be produced. Our results demonstrate that the recombinant cold-adapted enzyme is produced in P. haloplanktis TAC125 in good yields and in a completely soluble and catalytically competent form. Moreover, an evaluation of optimal induction conditions for protein production was also carried out in two consecutive steps: (1) definition of the optimal cellular growth phase in which the gene expression has to be induced; (2) definition of the optimal inducer concentration that has to be added in the growth medium. © 2012 Springer Science+Business Media, LLC.
CITATION STYLE
Rippa, V., Papa, R., Giuliani, M., Pezzella, C., Parrilli, E., Tutino, M. L., … Duilio, A. (2012). Regulated recombinant protein production in the antarctic bacterium Pseudoalteromonas haloplanktis TAC125. Methods in Molecular Biology, 824, 203–218. https://doi.org/10.1007/978-1-61779-433-9_10
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