Design of a novel switchable antibody display system in Pichia pastoris

Gätjen, Dominic; Tomszak, Florian; Dettmann, Johann-Christoph; Droste, Miriam; Nölle, Volker; Wieczorek, Marek

doi:10.1007/s00253-022-12108-5

Cited by 6 publications

(6 citation statements)

References 68 publications

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“…Although this review outlines molecular display technologies only for S. cerevisiae , other yeasts such as Komagataella pastoris ( Pichia pastoris ) and Yarrow lipolitica have also been developed as possible hosts for recombinant protein production [ 134 , 135 ]. For example, the lipase Lip2 was displayed on the surface of Y. lipolitica [ 136 ], and the investigation showed that Lip2 exhibited higher stability in organic acids and detergents, such as EDTA, SDS, dimethyl sulfoxide, and Tween 80.…”

Section: Discussionmentioning

confidence: 99%

Progress of Molecular Display Technology Using Saccharomyces cerevisiae to Achieve Sustainable Development Goals

Shibasaki

Ueda

2023

Microorganisms

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In the long history of microorganism use, yeasts have been developed as hosts for producing biologically active compounds or for conventional fermentation. Since the introduction of genetic engineering, recombinant proteins have been designed and produced using yeast or bacterial cells. Yeasts have the unique property of expressing genes derived from both prokaryotes and eukaryotes. Saccharomyces cerevisiae is one of the well-studied yeasts in genetic engineering. Recently, molecular display technology, which involves a protein-producing system on the yeast cell surface, has been established. Using this technology, designed proteins can be displayed on the cell surface, and novel abilities are endowed to the host yeast strain. This review summarizes various molecular yeast display technologies and their principles and applications. Moreover, S. cerevisiae laboratory strains generated using molecular display technology for sustainable development are described. Each application of a molecular displayed yeast cell is also associated with the corresponding Sustainable Development Goals of the United Nations.

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Section: Discussionmentioning

confidence: 99%

Progress of Molecular Display Technology Using Saccharomyces cerevisiae to Achieve Sustainable Development Goals

Shibasaki

Ueda

2023

Microorganisms

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“…Promoters are usually selected from either natural resources or available modified promoters. For example, except for P AOX1 and P ADH3 , numerous types of natural promoters, i.e., methanol-induced P DAS1 [ 39 ], P DAS2 [ 39 ], and P FLD1 [ 40 ], rhamnose-induced P LAR3 [ 41 ], and constitutive promoters P TEF1 [ 42 ] and P GCW14 [ 43 ], have been reported to promote the strong expression of recombinant proteins. Furthermore, in addition to the modified promoter P AOXm , modifications are also made for P GAP [ 44 ] and P CAT1 [ 45 ], among others, to improve the expression yield.…”

Section: Discussionmentioning

confidence: 99%

Enhanced Expression of Alcohol Dehydrogenase I in Pichia pastoris Reduces the Content of Acetaldehyde in Wines

Geng,

Lin,

Zheng

et al. 2023

Microorganisms

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Acetaldehyde is an important carbonyl compound commonly detected in wines. A high concentration of acetaldehyde can affect the flavor of wines and result in adverse effects on human health. Alcohol dehydrogenase I (ADH1) in Saccharomyces cerevisiae catalyzes the reduction reaction of acetaldehyde into ethanol in the presence of cofactors, showing the potential to reduce the content of acetaldehyde in wines. In this study, ADH1 was successfully expressed in Pichia pastoris GS115 based on codon optimization. Then, the expression level of ADH1 was enhanced by replacing its promoter with optimized promoters and increasing the copy number of the expression cassette, with ADH1 being purified using nickel column affinity chromatography. The enzymatic activity of purified ADH1 reached 605.44 ± 44.30 U/mg. The results of the effect of ADH1 on the content of acetaldehyde in wine revealed that the acetaldehyde content of wine samples was reduced from 168.05 ± 0.55 to 113.17 ± 6.08 mg/L with the addition of 5 mM NADH and the catalysis of ADH1, and from 135.53 ± 4.08 to 52.89 ± 2.20 mg/L through cofactor regeneration. Our study provides a novel approach to reducing the content of acetaldehyde in wines through enzymatic catalysis.

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“…Besides HR efficiency, the vector carrying sgRNA expression cassette could be eliminated easily, benefiting the editing of other genes ( Yang et al, 2020 ). It is necessary to identify two genetic elements, namely, the centromere and the autonomously replicating sequence (ARS), to engineer a steady episomal plasmid in a microorganism without carrying a plasmid ( Piva et al, 2020 ; Schultz et al, 2021 ; Gatjen et al, 2022 ). Several recently identified ARS enable maintenance of episomal plasmids and efficient expression of recombinant proteins ( Camattari et al, 2016 ; Schwarzhans et al, 2017 ; Nakamura et al, 2018 ; Gatjen et al, 2022 ).…”

Section: Host Strain Engineeringmentioning

confidence: 99%

“…It is necessary to identify two genetic elements, namely, the centromere and the autonomously replicating sequence (ARS), to engineer a steady episomal plasmid in a microorganism without carrying a plasmid ( Piva et al, 2020 ; Schultz et al, 2021 ; Gatjen et al, 2022 ). Several recently identified ARS enable maintenance of episomal plasmids and efficient expression of recombinant proteins ( Camattari et al, 2016 ; Schwarzhans et al, 2017 ; Nakamura et al, 2018 ; Gatjen et al, 2022 ). For instance, the episomal plasmids can be maintained in P. pastoris by a 452 bp DNA sequence (pan ARS) recovered from Kluyveromyces lactis ( Camattari et al, 2016 ).…”

Section: Host Strain Engineeringmentioning

confidence: 99%

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

Pan

Yang²,

Wu³

et al. 2022

Front. Microbiol.

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Pichia pastoris (syn. Komagataella spp.) has attracted extensive attention as an efficient platform for recombinant protein (RP) production. For obtaining a higher protein titer, many researchers have put lots of effort into different areas and made some progress. Here, we summarized the most recent advances of the last 5 years to get a better understanding of its future direction of development. The appearance of innovative genetic tools and methodologies like the CRISPR/Cas9 gene-editing system eases the manipulation of gene expression systems and greatly improves the efficiency of exploring gene functions. The integration of novel pathways in microorganisms has raised more ideas of metabolic engineering for enhancing RP production. In addition, some new opportunities for the manufacture of proteins have been created by the application of novel mathematical models coupled with high-throughput screening to have a better overview of bottlenecks in the biosynthetic process.

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Design of a novel switchable antibody display system in Pichia pastoris

Cited by 6 publications

References 68 publications

Progress of Molecular Display Technology Using Saccharomyces cerevisiae to Achieve Sustainable Development Goals

Progress of Molecular Display Technology Using Saccharomyces cerevisiae to Achieve Sustainable Development Goals

Enhanced Expression of Alcohol Dehydrogenase I in Pichia pastoris Reduces the Content of Acetaldehyde in Wines

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

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