Pichia pastoris versus Saccharomyces cerevisiae: a case study on the recombinant production of human granulocyte-macrophage colony-stimulating factor

Tran, Anh Tu; Nguyen, Thanh Thao; Nguyen, Cong Thuan; Huynh-Thi, Xuan Mai; Nguyen, Cao T.D.; Trinh, Minh Thuong; Tran, Linh Thuoc; Cartwright, Stephanie P.; Bill, Roslyn M.; Tran‐Van, Hieu

doi:10.1186/s13104-017-2471-6

Cited by 26 publications

(12 citation statements)

References 15 publications

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“…Delic and colleagues highlighted differences between yeast species by comparing their canonical protein secretion pathway with S. cerevisiae [ 33 , 88 ]. In several cases, the secretion yields of the recombinant products by K. phaffii often surpass those achieved with S. cerevisiae , which can also result from higher biomass accumulation [ 33 , 89 , 90 , 91 ]. Approximately 10% of the total genes in K. phaffii ’s genome is predicted to have a role in the secretory pathway, comprising those marked to (i) ER, (ii) protein folding, (iii) glycosylation, (iv) proteolytic processing, (v) ER-associated degradation (ERAD) pathway, (vi) Golgi apparatus, (vii) SNAREs, and (viii) others involved in vesicle-mediated transport [ 33 , 88 ].…”

Section: Protein Secretion: Bottlenecks Of the Secretory Pathwaymentioning

confidence: 99%

Industrial Production of Proteins with Pichia pastoris—Komagataella phaffii

Barone

Emmerstorfer-Augustin

Biundo

et al. 2023

Biomolecules

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Since the mid-1960s, methylotrophic yeast Komagataella phaffii (previously described as Pichia pastoris) has received increasing scientific attention. The interest for the industrial production of proteins for different applications (e.g., feed, food additives, detergent, waste treatment processes, and textile) is a well-consolidated scientific topic, and the importance for this approach is rising in the current era of environmental transition in human societies. This review aims to summarize fundamental and specific information in this scientific field. Additionally, an updated description of the relevant products produced with K. phaffii at industrial levels by a variety of companies—describing how the industry has leveraged its key features, from products for the ingredients of meat-free burgers (e.g., IMPOSSIBLE™ FOODS, USA) to diabetes therapeutics (e.g., Biocon, India)—is provided. Furthermore, active patents and the typical workflow for industrial protein production with this strain are reported.

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Section: Protein Secretion: Bottlenecks Of the Secretory Pathwaymentioning

confidence: 99%

Industrial Production of Proteins with Pichia pastoris—Komagataella phaffii

Barone

Emmerstorfer-Augustin

Biundo

et al. 2023

Biomolecules

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“… 2011 ; Tran et al . 2017 ). Similar to S. cerevisiae , about 10% of the total genes in the P. pastoris genome are predicted to have a function in the secretory pathway, including genes annotated to ER, protein folding, glycosylation, proteolytic processing, ERAD, Golgi, the vacuole, SNAREs and other genes involved in vesicle‐mediated transport (Delic et al .…”

Section: Protein Secretionmentioning

confidence: 99%

What makes Komagataella phaffii non-conventional?

Ata

Ergün

Fickers

et al. 2021

FEMS Yeast Research

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The important industrial protein production host Komagataella phaffii (syn Pichia pastoris) is classified as a non-conventional yeast. But what exactly makes K. phaffii non-conventional? In this review we set out to address the main differences to the ‘conventional’ yeast Saccharomyces cerevisiae, but also pinpoint differences to other non-conventional yeasts used in biotechnology. Apart from its methylotrophic lifestyle, K. phaffii is a Crabtree-negative yeast species. But even within the methylotrophs, K. phaffii possesses distinct regulatory features such as glycerol-repression of the methanol-utilization pathway or the lack of nitrate assimilation. Rewiring of the transcriptional networks regulating carbon (and nitrogen) source utilization clearly contributes to our understanding of genetic events occurring during evolution of yeast species. The mechanisms of mating-type switching and the triggers of morphogenic phenotypes represent further examples for how K. phaffii is distinguished from the model yeast S. cerevisiae. With respect to heterologous protein production, K. phaffii features high secretory capacity but secretes only low amounts of endogenous proteins. Different to S. cerevisiae, the Golgi apparatus of K. phaffii is stacked like in mammals. While it is tempting to speculate that Golgi architecture is correlated to the high secretion levels or the different N-glycan structures observed in K. phaffii, there is recent evidence against this. We conclude that K. phaffii is a yeast with unique features that has a lot of potential to explore both fundamental research questions and industrial applications.

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“…Commercial kits for this expression system are available [8]. Moreover, less extensive glycosylation in P. pastoris than in S. cerevisiae creates additional benefits in reducing immunogenicity [9].…”

Section: Introductionmentioning

confidence: 99%

Evaluating Pulses and Modified Fed-batch Feeding of Methanol to Increase Expression Level of Human Insulin Precursor in Pichia Pastoris High-Density Cultivation

Putro¹,

Nurdiani²,

Hariyatun³

et al. 2022

International Journal on Advanced Science, Engineering and Information Technology

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Production of sufficient insulin at a more affordable price is necessary. The increase in the number of people living with diabetes puts more burden on healthcare and the economy. P. pastoris is a promising host to produce human insulin precursors at a high yield in minimal medium and secretes low levels of endogenous protein impurities. Production of the precursor involves several parameters, including glycerol concentration, culture density, methanol concentration, and medium composition. This study evaluated the effect of those parameters on protein expression in the flask culture. Subsequently, fermentation in the bioreactor was carried on according to the information obtained from flask culture. Methanol feeding to induce protein expression was undertaken by pulses and fed-batch modes. The fed-batch method was modified from a standard technique by incorporating constant flow rates with variable feed concentrations. Cell density was determined based on optical density measurement at 600 nm and dry cell weight. Tricine SDS-PAGE and reversed-phase HPLC conducted protein analysis. The pulse feeding produced higher precursor concentrations at ~445 mg/L than modified fed-batch feeding at ~267 mg/L. However, the modified fed-batch feeding can be an alternative to producing human insulin precursors when a standard fed-batch feeding with variable flow rates and 100% (v/v) methanol feed is difficult to apply.

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Pichia pastoris versus Saccharomyces cerevisiae: a case study on the recombinant production of human granulocyte-macrophage colony-stimulating factor

Cited by 26 publications

References 15 publications

Industrial Production of Proteins with Pichia pastoris—Komagataella phaffii

Industrial Production of Proteins with Pichia pastoris—Komagataella phaffii

What makes Komagataella phaffii non-conventional?

Evaluating Pulses and Modified Fed-batch Feeding of Methanol to Increase Expression Level of Human Insulin Precursor in Pichia Pastoris High-Density Cultivation

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