Advanced Strategies for Food-Grade Protein Production: A New E. coli/Lactic Acid Bacteria Shuttle Vector for Improved Cloning and Food-Grade Expression

Tagliavia, Marcello; Nicosia, Aldo

doi:10.3390/microorganisms7050116

Cited by 22 publications

(16 citation statements)

References 50 publications

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“…Alternatively, a shuttle vector with a removable selection marker allows cloning in E. coli and protein expression in LAB. The E. coli cassette can be easily excised from the selected recombinant plasmid, and the resulting antibiotic-selection-marker-free vector transformed into the final food-grade expression host L. lactis NZ3000 with lacF gene deletion [115].…”

Section: Use Of Homologous Dnamentioning

confidence: 99%

Safety Aspects of Genetically Modified Lactic Acid Bacteria

Plavec

Berlec

2020

Microorganisms

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Lactic acid bacteria (LAB) have a long history of use in the food industry. Some species are part of the normal human microbiota and have beneficial properties for human health. Their long-standing use and considerable biotechnological potential have led to the development of various systems for their engineering. Together with novel approaches such as CRISPR-Cas, the established systems for engineering now allow significant improvements to LAB strains. Nevertheless, genetically modified LAB (GM-LAB) still encounter disapproval and are under extensive regulatory requirements. This review presents data on the prospects for LAB to obtain ‘generally recognized as safe’ (GRAS) status. Genetic modification of LAB is discussed, together with problems that can arise from their engineering, including their dissemination into the environment and the spread of antibiotic resistance markers. Possible solutions that would allow the use of GM-LAB are described, such as biocontainment, alternative selection markers, and use of homologous DNA. The use of GM-LAB as cell factories in closed systems that prevent their environmental release is the least problematic aspect, and this is also discussed.

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Section: Use Of Homologous Dnamentioning

confidence: 99%

Safety Aspects of Genetically Modified Lactic Acid Bacteria

Plavec

Berlec

2020

Microorganisms

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“…E. coli expression systems are often used to produce exogenous protein on laboratory and industrial scales owing to the low cost, speed, and simplicity of cultivation [ 22 ]. E. coli strains DH5α and Rosetta (DE3) have been used for the cloning of genes and the expression of proteins.…”

Section: Host Strains For the Overexpression Of Target Proteinsmentioning

confidence: 99%

“…Co-overexpression of molecular chaperones is a potential method for avoiding the development of inclusion bodies. This technique is appealing; however, there is no certainty that chaperones can increase the solubility of recombinant proteins [ 22 , 139 ]. E. coli encode chaperones, some of which drive the process of protein folding, while others inhibit protein aggregation [ 134 ].…”

Section: Co-expression Of Chaperonesmentioning

confidence: 99%

Strategies for Optimizing the Production of Proteins and Peptides with Multiple Disulfide Bonds

Lee

Park

2020

Antibiotics

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Bacteria can produce recombinant proteins quickly and cost effectively. However, their physiological properties limit their use for the production of proteins in their native form, especially polypeptides that are subjected to major post-translational modifications. Proteins that rely on disulfide bridges for their stability are difficult to produce in Escherichia coli. The bacterium offers the least costly, simplest, and fastest method for protein production. However, it is difficult to produce proteins with a very large size. Saccharomyces cerevisiae and Pichia pastoris are the most commonly used yeast species for protein production. At a low expense, yeasts can offer high protein yields, generate proteins with a molecular weight greater than 50 kDa, extract signal sequences, and glycosylate proteins. Both eukaryotic and prokaryotic species maintain reducing conditions in the cytoplasm. Hence, the formation of disulfide bonds is inhibited. These bonds are formed in eukaryotic cells during the export cycle, under the oxidizing conditions of the endoplasmic reticulum. Bacteria do not have an advanced subcellular space, but in the oxidizing periplasm, they exhibit both export systems and enzymatic activities directed at the formation and quality of disulfide bonds. Here, we discuss current techniques used to target eukaryotic and prokaryotic species for the generation of correctly folded proteins with disulfide bonds.

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“…Lactic acid bacteria (LAB) as Streptococcus thermophilus are widely used for industrial food fermentation, but also generate great interest for their food-grade expression capabilities (proteins or metabolites) [15,16]. To do so, replicating plasmids are often used to introduce genetic material into LAB cells [17,18], but these plasmids need to be maintained into the cells.…”

Section: Introductionmentioning

confidence: 99%

“French Phage Network” Annual Conference—Fifth Meeting Report

Laumay

Chaïb

Linares

et al. 2020

Viruses

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Attracting about 100 participants, the fifth edition of our French Phages.fr annual conference was once more a success. This year’s conference took place at the Institute for Structural Biology on the European Electron and Photon Campus in Grenoble, 8–9 October 2019. Similar to previous years, our meeting gathered scientists mainly working in France, from academic labs and hospitals as well as from industry. We also had the pleasure of welcoming attendees from different European countries and even beyond. The conference was divided into four sessions: Ecology and Evolution, Phage Therapy and Biotechnology, Structure and Assembly and Phage–Host Interaction, each opened by a keynote lecture. The talks, selected from abstracts, gave the opportunity for young scientists (especially students and post-docs) to present their project and results in a friendly atmosphere. Poster sessions also favoured interactions and discussions between young researchers and more senior scientists. Here, we provide a summary of the topics developed during the conference.

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Advanced Strategies for Food-Grade Protein Production: A New E. coli/Lactic Acid Bacteria Shuttle Vector for Improved Cloning and Food-Grade Expression

Cited by 22 publications

References 50 publications

Safety Aspects of Genetically Modified Lactic Acid Bacteria

Safety Aspects of Genetically Modified Lactic Acid Bacteria

Strategies for Optimizing the Production of Proteins and Peptides with Multiple Disulfide Bonds

“French Phage Network” Annual Conference—Fifth Meeting Report

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