Effect of Methionine on Gene Expression in Komagataella phaffii Cells

Ianshina, Tatiana; Sidorin, Anton; Petrova, Kristina; Shubert, Maria; Makeeva, Anastasiya S.; Самбук, Е. В.; Govdi, Anastasia I.; Rumyantsev, Andrey M.; Падкина, М. В.

doi:10.3390/microorganisms11040877

Cited by 3 publications

(3 citation statements)

References 73 publications

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“…The practical importance of K. phaffii and its growing recognition among researchers allowed this yeast to become a new model organism in fundamental research [ 10 ]. “Omics” technologies (genomics [ 11 ], transcriptomics [ 12 , 13 ], proteomics [ 14 ], lipidomics [ 15 ] and metabolomics [ 16 ]) have greatly accelerated the use of K. phaffii .…”

Section: Introductionmentioning

confidence: 99%

Alternative PCR-Based Approaches for Generation of Komagataella phaffii Strains

Makeeva,

Muzaev,

Shubert

et al. 2023

Microorganisms

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Komagataella phaffii (Pichia pastoris) is a widely known microbial host for recombinant protein production and an emerging model organism in fundamental research. The development of new materials and techniques on this yeast improves heterologous protein synthesis. One of the most prominent ways to enhance protein production efficiency is to select K. phaffii strains with multiple expression cassettes and generate MutS strains using various vectors. In this study, we demonstrate approaches to expand the applications of pPICZ series vectors. Procedures based on PCR amplification and in vivo cloning allow rapid exchange of selectable markers. The combination of PCR amplification with split-marker-mediated transformation allows the development of K. phaffii MutS strains with two expression cassettes using pPICZ vectors. Both PCR-based approaches were applied to efficiently produce interleukin-2 mimetic Neo-2/15 in K. phaffii. The described techniques provide alternative ways to generate and improve K. phaffii strains without the need for obtaining new specific vectors or additional cloning of expression cassettes.

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Section: Introductionmentioning

confidence: 99%

Alternative PCR-Based Approaches for Generation of Komagataella phaffii Strains

Makeeva,

Muzaev,

Shubert

et al. 2023

Microorganisms

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“…Protein synthesis improvement in this yeast includes selection of optimal cultivation parameters [1, 2]. Therefore, much attention is paid to the influence of media components on physiology of this yeast [3][4][5].One of the essential media components is biotin. In yeast cells it plays a crucial role as a cofactor of enzymes, providing carboxylation reactions in lipo-, gluconeogenesis, and nitrogen metabolism.…”

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confidence: 99%

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confidence: 99%

Effect of biotin starvation on gene expression in industrially significant yeast Komagataella phaffii

Makeeva,

Sidorin,

Ishtuganova

et al. 2023

Ecological genetics

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Komagataella phaffii(Pichia pastoris) is known to be an excellent producer of recombinant proteins for industrial and research purposes. Protein synthesis improvement in this yeast includes selection of optimal cultivation parameters [1, 2]. Therefore, much attention is paid to the influence of media components on physiology of this yeast [3–5]. One of the essential media components is biotin. In yeast cells it plays a crucial role as a cofactor of enzymes, providing carboxylation reactions in lipo-, gluconeogenesis, and nitrogen metabolism.K. phaffiiis biotin auxotrophic organism unable to synthesize this vitaminde novo. Thus, it necessarily requires adding of biotin in the media. In this study, we analyzed the effect of biotin starvation on gene expression inK. phaffiicells during its growth on methanol- and glycerin-containing media. These carbon sources are the most commonly used in standard protocols for recombinant protein biosynthesis inK. phaffii. It was shown, that biotin starvation cell response significantly depends on carbon source. In glycerol-containing media biotin deficiency enhanced transcription of genes involved in biotin and thiamine metabolism, glyoxylate cycle, synthesis of acetyl-CoA in cytoplasm and its carnitine-mediated transport into mitochondria. Genes involved in biosynthesis of lipids and glucose were repressed in media with glycerol. In methanol-containing media the biotin deficiency effect was more pronounced and led to repression of numerous genes involved in protein and amino acids synthesis and activation of cell response to oxidative stress. The obtained results are thought to be important for optimizing the culture conditions in theK. phaffiiexpression systems.

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Genetic Collections of St. Petersburg University

Andreeva,

Burlakovskiy,

Buzovkina

et al. 2023

BioComm

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Bioresource collections represent a unique source of biological diversity for research in genetics and related disciplines. The Department of Genetics and Biotechnology of St. Petersburg State University is the oldest department of genetics in Russia, founded in 1919. Throughout the entire period of development, the geneticists of St. Petersburg University have collected unique forms of plants, animals and microorganisms, on which their research was based. Many of these studies including regulation of translation termination in yeast, amyloids and prions of different organisms, genetic mapping of valuable morphological and biochemical traits to create first rye chromosome maps, and several aspects of transcription regulation in plants, had a significant novelty. The most active accumulation of collections of genetic resources at St. Petersburg State University started in the 1950-1970s when important scientific directions in the genetics of microorganisms, plants and animals, many of which continue today, were established at the department. Genetic collections are actively used in educational work for teaching dozens of educational courses. Currently, the interdisciplinary genetic collections of St. Petersburg State University consist of seven sections including genetic collections of rye, radish, garden pea, Chlamydomonas algae, Saccharomyces yeast and plasmids, Komagataella yeast, Drosophila fly. This review describes in detail the collections of the Department of Genetics and Biotechnology of St. Petersburg State University and discusses their current state, application and development prospects.

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Effect of Methionine on Gene Expression in Komagataella phaffii Cells

Cited by 3 publications

References 73 publications

Alternative PCR-Based Approaches for Generation of Komagataella phaffii Strains

Alternative PCR-Based Approaches for Generation of Komagataella phaffii Strains

Effect of biotin starvation on gene expression in industrially significant yeast Komagataella phaffii

Genetic Collections of St. Petersburg University

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