Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 99, Issue 14, Pages 5863-5874Publisher
SPRINGER
DOI: 10.1007/s00253-014-6328-9
Keywords
Recombinant protein; Expression systems; Escherichia coli; Pseudoalteromonas haloplanktis TAC125; Human alpha-galactosidase A; Fabry's disease
Categories
Funding
- European Union [ERANET-IB08-007]
- national project [EUI2008-03610]
- Universitat Autonoma de Barcelona [EME2007-08]
- Antartide
- MIUR Azioni Integrate Italia-Spagna [Prot. IT10LECLM9]
- MINECO [IT2009-0021]
- AGAUR [2009SGR-108]
- Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN, Spain), an initiative - VI National RDi Plan
- Iniciativa Ingenio, Consolider Program, CIBER Actions
- Instituto de Salud Carlos III
- ISCIII
- ICREA ACADEMIA award (Catalonia, Spain)
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Obtaining high levels of pure proteins remains the main bottleneck of many scientific and biotechnological studies. Among all the available recombinant expression systems, Escherichia coli facilitates gene expression by its relative simplicity, inexpensive and fast cultivation, well-known genetics and the large number of tools available for its biotechnological application. However, recombinant expression in E. coli is not always a straightforward procedure and major obstacles are encountered when producing many eukaryotic proteins and especially membrane proteins, linked to missing posttranslational modifications, proteolysis and aggregation. In this context, many conventional and unconventional eukaryotic hosts are under exploration and development, but in some cases linked to complex culture media or processes. In this context, alternative bacterial systems able to overcome some of the limitations posed by E. coli keeping the simplicity of prokaryotic manipulation are currently emerging as convenient hosts for protein production. We have comparatively produced a difficult-to-express human protein, the lysosomal enzyme alpha-galactosidase A (hGLA) in E. coli and in the psychrophilic bacterium Pseudoalteromonas haloplanktis TAC125 cells (P. haloplanktis TAC125). While in E. coli the production of active hGLA was unreachable due to proteolytic instability and/or protein misfolding, the expression of hGLA gene in P. haloplanktis TAC125 allows obtaining active enzyme. These results are discussed in the context of emerging bacterial systems for protein production that represent appealing alternatives to the regular use of E. coli and also of more complex eukaryotic systems.
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