The new strategies to overcome challenges in protein production in bacteria

Lipońska, Anna; Ousalem, Farès; Aalberts, Daniel P.; Hunt, J.F.; Boël, Grégory

doi:10.1111/1751-7915.13338

Cited by 10 publications

(8 citation statements)

References 25 publications

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“…Moreover, additional challenges include mimicking eukaryotic post-translational modifications (such as glycosylation) [11], production of harmful endotoxins (E. coli) [12], reduced cell viability resulting from unwanted by-products [13], and low protein yields [14]. To tackle these obstacles, different recombinant DNA technologies, including manipulation of gene expression control [15][16][17], directing proper bond formation and protein folding [18,19], interfering host metabolic pathways [20,21], random or directed evolutions of bacterial strains or enzymes [22], and series of other methods have been systematically employed (summarized in [23]). While these engineering strategies already provide practical solutions for the expression of various protein forms, the engineered expression systems can further be optimized to achieve a better performance.…”

Section: Introductionmentioning

confidence: 99%

Engineering Biology to Construct Microbial Chassis for the Production of Difficult-to-Express Proteins

Kim

Choe

Lee

et al. 2020

IJMS

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A large proportion of the recombinant proteins manufactured today rely on microbe-based expression systems owing to their relatively simple and cost-effective production schemes. However, several issues in microbial protein expression, including formation of insoluble aggregates, low protein yield, and cell death are still highly recursive and tricky to optimize. These obstacles are usually rooted in the metabolic capacity of the expression host, limitation of cellular translational machineries, or genetic instability. To this end, several microbial strains having precisely designed genomes have been suggested as a way around the recurrent problems in recombinant protein expression. Already, a growing number of prokaryotic chassis strains have been genome-streamlined to attain superior cellular fitness, recombinant protein yield, and stability of the exogenous expression pathways. In this review, we outline challenges associated with heterologous protein expression, some examples of microbial chassis engineered for the production of recombinant proteins, and emerging tools to optimize the expression of heterologous proteins. In particular, we discuss the synthetic biology approaches to design and build and test genome-reduced microbial chassis that carry desirable characteristics for heterologous protein expression.

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Section: Introductionmentioning

confidence: 99%

Engineering Biology to Construct Microbial Chassis for the Production of Difficult-to-Express Proteins

Kim

Choe

Lee

et al. 2020

IJMS

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“…Two strategies can be applied to alleviate the host burden: heterologous gene codon optimization and supplementation of rare tRNAs. The former not only requires significant experimental resources, but also results in heavy competition for the internal tRNA pool, placing a heavier burden on the host [67]. Conversely, the appropriate introduction of rare codons can improve the yield and solubility of RPs and reduce the host burden [68,69].…”

Section: Balancing Cell Growth and Recombinant Protein Productionmentioning

confidence: 99%

Strategies for efficient production of recombinant proteins in Escherichia coli: alleviating the host burden and enhancing protein activity

et al. 2022

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Escherichia coli, one of the most efficient expression hosts for recombinant proteins (RPs), is widely used in chemical, medical, food and other industries. However, conventional expression strains are unable to effectively express proteins with complex structures or toxicity. The key to solving this problem is to alleviate the host burden associated with protein overproduction and to enhance the ability to accurately fold and modify RPs at high expression levels. Here, we summarize the recently developed optimization strategies for the high-level production of RPs from the two aspects of host burden and protein activity. The aim is to maximize the ability of researchers to quickly select an appropriate optimization strategy for improving the production of RPs.

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“…While the repetitive primary structure of PBMs can complicate in vitro DNA synthesis and in vivo stability, these challenges can be largely overcome by using algorithms to design DNA sequences that balance codon usage and minimize repetition. More problematically, while protein expression from a bacterial host is a seemingly well-established task, 20 many PBMs are of eukaryotic origin 5,6 and lead to assembled protein structures orders of magnitude larger than any bacterial cell. This is the case for spider silk fibers, which are also hard to synthesize due to metabolic burden and genetic instability caused by repetitions.…”

Section: Challenge 1: Controlling Secretion and Assembly To Deliver Engineerable Protein-based Elmsmentioning

confidence: 99%

Bottom-up approaches to engineered living materials: Challenges and future directions

Molinari

Tesoriero

Ajo‐Franklin

2021

Matter

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Biomaterials made by living systems, while very diverse, generally share similar organization: different lineages of cells are precisely patterned within a matrix to create a hierarchically defined structure. This meticulous organization allows for their multifunctional properties, which often exceed those of traditional materials. For these reasons, there is growing interest in creating engineered living materials (ELMs) that will have capabilities similar to those of natural living materials yet with tailored functions. This review aims to highlight technologies still missing from the field of ELMs, which currently prevents more impactful applications. We briefly review the existing literature and identify challenges in designing novel protein-based and non-ribosomally synthesized matrix elements, producing and incorporating biominerals, and improving critical material properties such as lifespan, spatial patterning, and cell-cell communication. We also discuss the interplay between these challenges and the need for the development of new chassis and corresponding genetic toolboxes. By overcoming these obstacles, we come ever closer to unlocking the potential and versatility of biomaterials to create designer ELMs.Protein-based materials (PBMs) are ubiquitous in nature and have often been studied and explored for commercial applications-especially in the biomedical field.

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The new strategies to overcome challenges in protein production in bacteria

Abstract: Recombinant proteins are essential for biotechnology. Here we review some of the key points for improving the production of heterologous proteins, and what can be the future of the field.

Cited by 10 publications

References 25 publications

Engineering Biology to Construct Microbial Chassis for the Production of Difficult-to-Express Proteins

Engineering Biology to Construct Microbial Chassis for the Production of Difficult-to-Express Proteins

Strategies for efficient production of recombinant proteins in Escherichia coli: alleviating the host burden and enhancing protein activity

Bottom-up approaches to engineered living materials: Challenges and future directions

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