Current advances of Pichia pastoris as cell factories for production of recombinant proteins

Pan, Yingjie; Yang, Jing; Wu, Jianping; Yang, Lirong; Hao, Fang

doi:10.3389/fmicb.2022.1059777

Cited by 32 publications

(19 citation statements)

References 139 publications

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“…This yeast has become an organism extensively used for heterologous protein expression. This popularity derives from a number of advantages, such as the ability to grow at high cellular densities, efficient mechanisms for secretion of produced proteins to the medium, low level of secreted endogenous proteins (which facilitates purification of heterologous secreted products), and a more suitable pattern of glycosylation of secreted proteins [ 17 , 18 ]. Heterologous protein expression in this organism is very often driven from the powerful AOX1 promoter, required for methanol utilization [ 19 ].…”

Section: Introductionmentioning

confidence: 99%

Transcriptomic profiling of the yeast Komagataella phaffii in response to environmental alkalinization

et al. 2023

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Background Adaptation to alkalinization of the medium in fungi involves an extensive remodeling of gene expression. Komagataella phaffii is an ascomycetous yeast that has become an organism widely used for heterologous protein expression. We explore here the transcriptional impact of moderate alkalinization in this yeast, in search of suitable novel promoters able to drive transcription in response to the pH signal. Results In spite of a minor effect on growth, shifting the cultures from pH 5.5 to 8.0 or 8.2 provokes significant changes in the mRNA levels of over 700 genes. Functional categories such as arginine and methionine biosynthesis, non-reductive iron uptake and phosphate metabolism are enriched in induced genes, whereas many genes encoding iron-sulfur proteins or members of the respirasome were repressed. We also show that alkalinization is accompanied by oxidative stress and we propose this circumstance as a common trigger of a subset of the observed changes. PHO89, encoding a Na+/Pi cotransporter, appears among the most potently induced genes by high pH. We demonstrate that this response is mainly based on two calcineurin-dependent response elements located in its promoter, thus indicating that alkalinization triggers a calcium-mediated signal in K. phaffii. Conclusions This work defines in K. phaffii a subset of genes and diverse cellular pathways that are altered in response to moderate alkalinization of the medium, thus setting the basis for developing novel pH-controlled systems for heterologous protein expression in this fungus.

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Section: Introductionmentioning

confidence: 99%

Transcriptomic profiling of the yeast Komagataella phaffii in response to environmental alkalinization

et al. 2023

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“…K. phaffii was developed as a platform for SCP production by Philips Petroleum and later used as a basis for the development of a protein production system (Cregg et al 1985 ). The advantages of the K. phaffii production system comprise high folding efficiency, high-cell-density fermentation, a strong and highly regulated expression system, genetic stability, robust protein secretion, easy downstream processing, FDA GRAS status, and suitability for the application of useful genome engineering techniques including the recently developed CRISPR/Cas9 gene editing tool (Zhu et al 2019 ; Pan et al 2022 ). According to Pichia.com, a web platform for posting information about the K. phaffii production system, over 5000 proteins have now been successfully produced by this system (Schwarzhans et al 2017 ).…”

Section: Komagataella Phaffiimentioning

confidence: 99%

Recent progress on heterologous protein production in methylotrophic yeast systems

Tsuda,

Nonaka

2024

World J Microbiol Biotechnol

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Recombinant protein production technology is widely applied to the manufacture of biologics used as drug substances and industrial proteins such as recombinant enzymes and bioactive proteins. Various heterologous protein production systems have been developed using prokaryotic and eukaryotic hosts. Especially methylotrophic yeast in eukaryotic hosts is suggested to be particularly valuable because such systems have the following advantages: protein secretion into culture broth, eukaryotic quality control systems, a post-translational modification system, rapid growth, and established recombinant DNA tools and technologies such as strong promoters, effective selection markers, and gene knock-in and -out systems. Many methylotrophic yeasts such as the genera Candida, Ogataea, and Komagataella have been studied since methylotrophic yeast was first isolated in 1969. The methanol-consumption-related genes in methylotrophic yeast are strongly and strictly regulated under methanol-containing conditions. The well-regulated gene expression systems under the methanol-inducible gene promoter lead to the potential application of heterologous protein production in methylotrophic yeast. In this review, we describe the recent progress of heterologous protein production technology in methylotrophic yeast and introduce Ogataea minuta as an alternative production host as a substitute for K. phaffii and O. polymorpha.

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“…As one of the most widely used protein expression systems in current applications, the Pichia pastoris expression system has been extensively employed for expressing various exogenous proteins such as serum proteins, insulin, and epidermal growth factor, among others [1]. Among these, cell concentration and product concentration are crucial process variables in the fermentation process, and the real-time and accurate detection of their values are essential for improving production efficiency and optimizing control [2]. For the measurement of key biological parameters during the fermentation process, offline sampling and laboratory analysis are mostly adopted [3].…”

Section: Introductionmentioning

confidence: 99%

Modeling and Optimization of an Enhanced Soft Sensor for the Fermentation Process of Pichia pastoris

Wang,

Yu,

Wang

et al. 2024

Sensors

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This paper proposes a novel soft sensor modeling approach, MIC-TCA-INGO-LSSVM, to address the decline in performance of soft sensor models during the fermentation process of Pichia pastoris, caused by changes in working conditions. Initially, the transfer component analysis (TCA) method is utilized to minimize the differences in data distribution across various working conditions. Subsequently, a least squares support vector machine (LSSVM) model is constructed using the dataset adapted by TCA, and strategies for improving the northern goshawk optimization (INGO) algorithm are proposed to optimize the parameters of the LSSVM model. Finally, to further enhance the model’s generalization ability and prediction accuracy, considering the transfer of knowledge from multiple-source working conditions, a sub-model weighted ensemble scheme is proposed based on the maximum information coefficient (MIC) algorithm. The proposed soft sensor model is employed to predict cell and product concentrations during the fermentation process of Pichia pastoris. Simulation results indicate that the RMSE of the INGO-LSSVM model in predicting cell and product concentrations is reduced by 47.3% and 42.1%, respectively, compared to the NGO-LSSVM model. Additionally, TCA significantly enhances the model’s adaptability when working conditions change. Moreover, the soft sensor model based on TCA and the MIC-weighted ensemble method achieves a reduction of 41.6% and 31.3% in the RMSE for predicting cell and product concentrations, respectively, compared to the single-source condition transfer model TCA-INGO-LSSVM. These results demonstrate the high reliability and predictive performance of the proposed soft sensor method under varying working conditions.

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Current advances of Pichia pastoris as cell factories for production of recombinant proteins

Cited by 32 publications

References 139 publications

Transcriptomic profiling of the yeast Komagataella phaffii in response to environmental alkalinization

Transcriptomic profiling of the yeast Komagataella phaffii in response to environmental alkalinization

Recent progress on heterologous protein production in methylotrophic yeast systems

Modeling and Optimization of an Enhanced Soft Sensor for the Fermentation Process of Pichia pastoris

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