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PubMed 22160900


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Automatically associated channels: Kv2.1



Title: Regulated recombinant protein production in the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125.

Authors: Valentina Rippa, Rosanna Papa, Maria Giuliani, Cinzia Pezzella, Ermenegilda Parrilli, Maria Luisa Tutino, Gennaro Marino, Angela Duilio

Journal, date & volume: Methods Mol. Biol., 2012 , 824, 203-18

PubMed link: http://www.ncbi.nlm.nih.gov/pubmed/22160900


Abstract
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.