scholarly journals Lactococcus lactis, an Efficient Cell Factory for Recombinant Protein Production and Secretion

2007 ◽  
Vol 14 (1-3) ◽  
pp. 48-58 ◽  
Author(s):  
E. Morello ◽  
L.G. Bermúdez-Humarán ◽  
D. Llull ◽  
V. Solé ◽  
N. Miraglio ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Lactococcus Lactis ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Cell Factory

scholarly journals Bioprocess performance analysis of novel methanol-independent promoters for recombinant protein production with Pichia pastoris

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Javier Garrigós-Martínez ◽  
Kiira Vuoristo ◽  
Miguel Angel Nieto-Taype ◽  
Juha Tähtiharju ◽  
Jaana Uusitalo ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Recombinant Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Expression System ◽  
Cell Factory ◽  
The Novel ◽  
Expression Systems ◽  
Fed Batch ◽  
Model Protein

Abstract Background Pichia pastoris is a powerful and broadly used host for recombinant protein production (RPP), where past bioprocess performance has often been directed with the methanol regulated AOX1 promoter (PAOX1), and the constitutive GAP promoter (PGAP). Since promoters play a crucial role in an expression system and the bioprocess efficiency, innovative alternatives are constantly developed and implemented. Here, a thorough comparative kinetic characterization of two expression systems based on the commercial PDF and UPP promoters (PPDF, PUPP) was first conducted in chemostat cultures. Most promising conditions were subsequently tested in fed-batch cultivations. These new alternatives were compared with the classical strong promoter PGAP, using the Candida antarctica lipase B (CalB) as model protein for expression system performance. Results Both the PPDF and PUPP-based expression systems outperformed similar PGAP-based expression in chemostat cultivations, reaching ninefold higher specific production rates (qp). CALB transcription levels were drastically higher when employing the novel expression systems. This higher expression was also correlated with a marked upregulation of unfolded protein response (UPR) related genes, likely from an increased protein burden in the endoplasmic reticulum (ER). Based on the chemostat results obtained, best culture strategies for both PPDF and PUPP expression systems were also successfully implemented in 15 L fed-batch cultivations where qp and product to biomass yield (YP/X*) values were similar than those obtained in chemostat cultivations. Conclusions As an outcome of the macrokinetic characterization presented, the novel PPDF and PUPP were observed to offer much higher efficiency for CalB production than the widely used PGAP-based methanol-free alternative. Thus, both systems arise as highly productive alternatives for P. pastoris-based RPP bioprocesses. Furthermore, the different expression regulation patterns observed indicate the level of gene expression can be adjusted, or tuned, which is interesting when using Pichia pastoris as a cell factory for different products of interest.

Recent Advances in Recombinant Protein Production by Bacillus subtilis

2020 ◽  
Vol 11 (1) ◽  
pp. 295-318 ◽  
Author(s):  
Kang Zhang ◽  
Lingqia Su ◽  
Jing Wu
Keyword(s):  
Bacillus Subtilis ◽  
Recombinant Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Genetic Manipulation ◽  
Microbial Cell ◽  
Secretory Proteins ◽  
Cell Factory ◽  
Microbial Cell Factory ◽  
Recent Advances

Bacillus subtilis has become a widely used microbial cell factory for the production of recombinant proteins, especially those associated with foods and food processing. Recent advances in genetic manipulation and proteomic analysis have been used to greatly improve protein production in B. subtilis. This review begins with a discussion of genome-editing technologies and application of the CRISPR–Cas9 system to B. subtilis. A summary of the characteristics of crucial legacy strains is followed by suggestions regarding the choice of origin strain for genetic manipulation. Finally, the review analyzes the genes and operons of B. subtilis that are important for the production of secretory proteins and provides suggestions and examples of how they can be altered to improve protein production. This review is intended to promote the engineering of this valuable microbial cell factory for better recombinant protein production.

scholarly journals Recombinant protein secretion by Bacillus subtilis and Lactococcus lactis: pathways, applications, and innovation potential

2021 ◽  
Author(s):  
Jolanda Neef ◽  
Jan Maarten van Dijl ◽  
Girbe Buist
Keyword(s):  
Bacillus Subtilis ◽  
Recombinant Protein ◽  
Lactococcus Lactis ◽  
Protein Secretion ◽  
Protein Production ◽  
Cell Factory ◽  
Efficient Production ◽  
High Quality ◽  
Gram Positive ◽  
Secretion Pathway

Abstract Secreted recombinant proteins are of great significance for industry, healthcare and a sustainable bio-based economy. Consequently, there is an ever-increasing need for efficient production platforms to deliver such proteins in high amounts and high quality. Gram-positive bacteria, particularly bacilli such as Bacillus subtilis, are favored for the production of secreted industrial enzymes. Nevertheless, recombinant protein production in the B. subtilis cell factory can be very challenging due to bottlenecks in the general (Sec) secretion pathway as well as this bacterium’s intrinsic capability to secrete a cocktail of highly potent proteases. This has placed another Gram-positive bacterium, Lactococcus lactis, in the focus of attention as an alternative, non-proteolytic, cell factory for secreted proteins. Here we review our current understanding of the secretion pathways exploited in B. subtilis and L. lactis to deliver proteins from their site of synthesis, the cytoplasm, into the fermentation broth. An advantage of this cell factory comparison is that it identifies opportunities for protein secretion pathway engineering to remove or bypass current production bottlenecks. Noteworthy new developments in cell factory engineering are the mini-Bacillus concept, highlighting potential advantages of massive genome minimization, and the application of thus far untapped ‘non-classical’ protein secretion routes. Altogether, it is foreseen that engineered lactococci will find future applications in the production of high-quality proteins at the relatively small pilot scale, while engineered bacilli will remain a favored choice for protein production in bulk.

scholarly journals Bioreactor-Scale Strategies for the Production of Recombinant Protein in the Yeast Yarrowia lipolytica

2019 ◽  
Vol 7 (2) ◽  
pp. 40 ◽  
Author(s):  
Marie Vandermies ◽  
Patrick Fickers
Keyword(s):  
Recombinant Protein ◽  
Yarrowia Lipolytica ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Genetic Manipulation ◽  
Research Field ◽  
Cell Factory ◽  
Cell Factories ◽  
A Cell ◽  
High Level

Recombinant protein production represents a multibillion-dollar market. Therefore, it constitutes an important research field both in academia and industry. The use of yeast as a cell factory presents several advantages such as ease of genetic manipulation, growth at high cell density, and the possibility of post-translational modifications. Yarrowia lipolytica is considered as one of the most attractive hosts due to its ability to metabolize raw substrate, to express genes at a high level, and to secrete protein in large amounts. In recent years, several reviews have been dedicated to genetic tools developed for this purpose. Though the construction of efficient cell factories for recombinant protein synthesis is important, the development of an efficient process for recombinant protein production in a bioreactor constitutes an equally vital aspect. Indeed, a sports car cannot drive fast on a gravel road. The aim of this review is to provide a comprehensive snapshot of process tools to consider for recombinant protein production in bioreactor using Y. lipolytica as a cell factory, in order to facilitate the decision-making for future strain and process engineering.

Continuous bleed recycling significantly increases recombinant protein production yield in perfusion cell cultures

2021 ◽  
Vol 169 ◽  
pp. 107966
Author(s):  
Jean-Marc Bielser ◽  
Mathieu Aeby ◽  
Stefania Caso ◽  
Anaïs Roulet ◽  
Hervé Broly ◽  
...  
Keyword(s):  
Recombinant Protein ◽  
Cell Cultures ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Production Yield
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