Abstract
BackgroundThe final aim of recombinant protein production is both to have a high specific production rate and a high product quality. It was already shown that using cold-adapted bacteria as host vectors, some "intractable" proteins can be efficiently produced at temperature as low as 4°C.ResultsA novel genetic system for the production and secretion of recombinant proteins in the Antarctic Gram-negative bacterium Pseudoalteromonas haloplanktis TAC125 was set up. This system aims at combining the low temperature recombinant product production with the advantages of extra-cellular protein targeting.The psychrophilic α-amylase from Pseudoalteromonas haloplanktis TAB23 was used as secretion carrier. Three chimerical proteins were produced by fusing intra-cellular proteins to C-terminus of the psychrophilic α-amylase and their secretion was analysed. Data reported in this paper demonstrate that all tested chimeras were translocated with a secretion yield always higher than 80%.ConclusionData presented here demonstrate that the "cold" gene-expression system is efficient since the secretion yield of tested chimeras is always above 80%. These secretion performances place the α-amylase derived secretion system amongst the best heterologous secretion systems in Gram-negative bacteria reported so far. As for the quality of the secreted passenger proteins, data presented suggest that the system also allows the correct disulphide bond formation of chimera components, secreting a fully active passenger.
Highlights
The final aim of recombinant protein production is both to have a high specific production rate and a high product quality
Microbial Cell Factories 2006, 5:40 http://www.microbialcellfactories.com/content/5/1/40 ing of a "difficult" product [3,4], a new expression system [5] was recently developed which implemented the use of the Antarctic Gram-negative bacterium Pseudoalteromonas haloplanktis TAC125 (P. haloplanktis TAC125) [6] as host for protein production
In this paper we report the setting up and use of a "cold" gene-expression system implemented for the secretion of recombinant proteins in P. haloplanktis TAC125
Summary
The final aim of recombinant protein production is both to have a high specific production rate and a high product quality. Either in the research community and biotechnology industry, Escherichia coli is the prokaryotic vector of choice for the high-level expression of proteins [1] This process sometimes results in the production of insoluble protein aggregates, incorrectly folded or nonfunctional proteins and proteins which may be degraded or contaminated with high levels of host-encoded proteins [2]. Microbial Cell Factories 2006, 5:40 http://www.microbialcellfactories.com/content/5/1/40 ing of a "difficult" product [3,4], a new expression system [5] was recently developed which implemented the use of the Antarctic Gram-negative bacterium Pseudoalteromonas haloplanktis TAC125 (P. haloplanktis TAC125) [6] as host for protein production By using such non-conventional system, some "intractable" proteins can be efficiently produced in soluble and active form at temperature as low as 4°C [7,8,9]. Protein secretion into the extra-cellular (outside) environment is the most desirable strategy; secreted proteins are not contaminated with other proteins and can be purified
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