Regulated recombinant protein production in the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125.
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;...
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Argelia Lorence
2012
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ftunivnapoliiris:oai:www.iris.unina.it:11588/430492 2024-06-23T07:47:45+00:00 Regulated recombinant protein production in the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125. RIPPA, VALENTINA PAPA, ROSANNA GIULIANI, MARIA PEZZELLA, Cinzia PARRILLI, ERMENEGILDA TUTINO, MARIA LUISA MARINO, GENNARO DUILIO, ANGELA Argelia Lorence Rippa, Valentina Papa, Rosanna Giuliani, Maria Pezzella, Cinzia Parrilli, Ermenegilda Tutino, MARIA LUISA Marino, Gennaro Duilio, Angela 2012 STAMPA http://hdl.handle.net/11588/430492 https://doi.org/10.1007/978-1-61779-433-9_10 eng eng Argelia Lorence country:USA place:New York info:eu-repo/semantics/altIdentifier/isbn/9781617794322 info:eu-repo/semantics/altIdentifier/wos/WOS:000303413800010 ispartofbook:Recombinant Gene Expression: Reviews and Protocols,Methods in Molecular Biology, volume:824 firstpage:203 lastpage:218 numberofpages:16 http://hdl.handle.net/11588/430492 doi:10.1007/978-1-61779-433-9_10 info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-84855968754 Cold-adapted bacteria - Regulated gene expression system - Pseudoalteromonas haloplanktis - Malate induction - Minimal medium info:eu-repo/semantics/bookPart 2012 ftunivnapoliiris https://doi.org/10.1007/978-1-61779-433-9_10 2024-06-10T14:58:48Z 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 b-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. Book Part Antarc* Antarctic IRIS Università degli Studi di Napoli Federico II Antarctic The Antarctic 203 218 |
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Open Polar |
collection |
IRIS Università degli Studi di Napoli Federico II |
op_collection_id |
ftunivnapoliiris |
language |
English |
topic |
Cold-adapted bacteria - Regulated gene expression system - Pseudoalteromonas haloplanktis - Malate induction - Minimal medium |
spellingShingle |
Cold-adapted bacteria - Regulated gene expression system - Pseudoalteromonas haloplanktis - Malate induction - Minimal medium RIPPA, VALENTINA PAPA, ROSANNA GIULIANI, MARIA PEZZELLA, Cinzia PARRILLI, ERMENEGILDA TUTINO, MARIA LUISA MARINO, GENNARO DUILIO, ANGELA Regulated recombinant protein production in the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125. |
topic_facet |
Cold-adapted bacteria - Regulated gene expression system - Pseudoalteromonas haloplanktis - Malate induction - Minimal medium |
description |
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 b-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. |
author2 |
Argelia Lorence Rippa, Valentina Papa, Rosanna Giuliani, Maria Pezzella, Cinzia Parrilli, Ermenegilda Tutino, MARIA LUISA Marino, Gennaro Duilio, Angela |
format |
Book Part |
author |
RIPPA, VALENTINA PAPA, ROSANNA GIULIANI, MARIA PEZZELLA, Cinzia PARRILLI, ERMENEGILDA TUTINO, MARIA LUISA MARINO, GENNARO DUILIO, ANGELA |
author_facet |
RIPPA, VALENTINA PAPA, ROSANNA GIULIANI, MARIA PEZZELLA, Cinzia PARRILLI, ERMENEGILDA TUTINO, MARIA LUISA MARINO, GENNARO DUILIO, ANGELA |
author_sort |
RIPPA, VALENTINA |
title |
Regulated recombinant protein production in the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125. |
title_short |
Regulated recombinant protein production in the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125. |
title_full |
Regulated recombinant protein production in the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125. |
title_fullStr |
Regulated recombinant protein production in the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125. |
title_full_unstemmed |
Regulated recombinant protein production in the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125. |
title_sort |
regulated recombinant protein production in the antarctic bacterium pseudoalteromonas haloplanktis tac125. |
publisher |
Argelia Lorence |
publishDate |
2012 |
url |
http://hdl.handle.net/11588/430492 https://doi.org/10.1007/978-1-61779-433-9_10 |
geographic |
Antarctic The Antarctic |
geographic_facet |
Antarctic The Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_relation |
info:eu-repo/semantics/altIdentifier/isbn/9781617794322 info:eu-repo/semantics/altIdentifier/wos/WOS:000303413800010 ispartofbook:Recombinant Gene Expression: Reviews and Protocols,Methods in Molecular Biology, volume:824 firstpage:203 lastpage:218 numberofpages:16 http://hdl.handle.net/11588/430492 doi:10.1007/978-1-61779-433-9_10 info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-84855968754 |
op_doi |
https://doi.org/10.1007/978-1-61779-433-9_10 |
container_start_page |
203 |
op_container_end_page |
218 |
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1802651904011927552 |