Pichia pastoris (Komagataella phaffii) as a Cost-Effective Tool for Vaccine Production for Low- and Middle-Income Countries (LMICs)

Magalhaes, Salome De Sa; Keshavarz-Moore, E

doi:10.3390/bioengineering8090119

Cited by 30 publications

(14 citation statements)

References 126 publications

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“…Development of facilities needed to produce mRNA therapeutics in massive quantities has been and is currently underway, with infrastructure capable of producing tens of millions of these products for vaccination and related applications [96]. The market value of vaccine production is also predicted to increase from 28 billion dollars reported in 2017 to over 80 billion dollars by 2027, with a CAGR of 8.7% [97]. With this increase in vaccine production worldwide, an updated view of vaccine manufacturing methods is required.…”

Section: Vaccinesmentioning

confidence: 99%

Biotechnology Applications of Cell-Free Expression Systems

Brookwell

Oza

Caschera

2021

Life

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Cell-free systems are a rapidly expanding platform technology with an important role in the engineering of biological systems. The key advantages that drive their broad adoption are increased efficiency, versatility, and low cost compared to in vivo systems. Traditionally, in vivo platforms have been used to synthesize novel and industrially relevant proteins and serve as a testbed for prototyping numerous biotechnologies such as genetic circuits and biosensors. Although in vivo platforms currently have many applications within biotechnology, they are hindered by time-constraining growth cycles, homeostatic considerations, and limited adaptability in production. Conversely, cell-free platforms are not hindered by constraints for supporting life and are therefore highly adaptable to a broad range of production and testing schemes. The advantages of cell-free platforms are being leveraged more commonly by the biotechnology community, and cell-free applications are expected to grow exponentially in the next decade. In this study, new and emerging applications of cell-free platforms, with a specific focus on cell-free protein synthesis (CFPS), will be examined. The current and near-future role of CFPS within metabolic engineering, prototyping, and biomanufacturing will be investigated as well as how the integration of machine learning is beneficial to these applications.

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Section: Vaccinesmentioning

confidence: 99%

Biotechnology Applications of Cell-Free Expression Systems

Brookwell

Oza

Caschera

2021

Life

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“…1,2 P. pastoris has already been proven repeatedly to allow high level recombinant protein production. 3,4,5,6 It is known from literature that recombinant protein production causes a metabolic burden on cellular metabolism. The phenotype of cells producing recombinant proteins can be a decreased growth rate, decreased biomass formation, and overall decreased cell fitness.…”

Section: Introductionmentioning

confidence: 99%

“…as the major players 1,2 . P. pastoris has already been proven repeatedly to allow high level recombinant protein production 3,4,5,6 …”

Section: Introductionmentioning

confidence: 99%

Customizing amino acid metabolism of Pichia pastoris for recombinant protein production

Rußmayer,

Buchetics,

Mattanovich

et al. 2023

Biotechnology Journal

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Amino acids are the building blocks of proteins. In this respect, a reciprocal effect of recombinant protein production on amino acid biosynthesis as well as the impact of the availability of free amino acids on protein production can be anticipated. In this study, the impact of engineering the amino acid metabolism on the production of recombinant proteins was investigated in the yeast Pichia pastoris (syn Komagataella phaffii). Based on comprehensive systems‐level analyses of the metabolomes and transcriptomes of different P. pastoris strains secreting antibody fragments, cell engineering targets were selected. Our working hypothesis that increasing intracellular amino acid levels could help unburden cellular metabolism and improve recombinant protein production was examined by constitutive overexpression of genes related to amino acid metabolism. In addition to twelve genes involved in specific amino acid biosynthetic pathways, the transcription factor GCN4 responsible for regulation of amino acid biosynthetic genes was overexpressed. The production of the used model protein, a secreted carboxylesterase (CES) from Sphingopyxis macrogoltabida, was increased by overexpression of pathway genes for alanine and for aromatic amino acids, and most pronounced, when overexpressing the regulator GCN4. The analysis of intracellular amino acid levels of selected clones indicated a direct linkage of improved recombinant protein production to the increased availability of intracellular amino acids. Finally, fed batch cultures showed that overexpression of GCN4 increased CES titers 2.6‐fold, while the positive effect of other amino acid synthesis genes could not be transferred from screening to bioreactor cultures.This article is protected by copyright. All rights reserved

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“…Yeast is a workhorse for the production of vaccines due to expression of different forms of antigens that can be scalable with reasonable cost. Among several yeast species, Pichia pastoris is considered one of the most promising options for heterologous protein production in vaccine development [ 8 ]. P. pastoris synthesizes high cell densities under a controlled environment and the cell lines are genetically stable in terms of the post translational modifications of protein in the endoplasmic reticulum (ER) and resistance to contamination.…”

mentioning

confidence: 99%

“…It has already been reported that more than 500 heterologous proteins have been successfully expressed using this host, with several of them being approved for human and commercial use. Human insulin (SuperMan5) and recombinant hepatitis C (HCV) are good examples of recombinant protein expressed in P. pastoris [ 8 ].…”

mentioning

confidence: 99%