Komagataella

Last updated

Komagataella
Pichia pastoris GS115 (Yang 2017).jpg
Komagataella phaffii [1] GS115
Scientific classification OOjs UI icon edit-ltr.svg
Domain: Eukaryota
Kingdom: Fungi
Division: Ascomycota
Class: Saccharomycetes
Order: Saccharomycetales
Family: Phaffomycetaceae
Genus: Komagataella
Y. Yamada, M. Matsuda, K. Maeda & Mikata, 1995
Species

See text

Komagataella is a methylotrophic yeast within the order Saccharomycetales. It was found in the 1960s as Pichia pastoris, with its feature of using methanol as a source of carbon and energy. [2] In 1995, P. pastoris was reassigned into the sole representative of genus Komagataella, becoming Komagataella phaffii. [3] Later studies have further distinguished new species in this genus, resulting in a total of 7 recognized species. [4] It is not uncommon to see the old name still in use in the context of protein production, as of 2023; [5] in less formal use, the yeast may confusingly be referred to as pichia.

Contents

After years of study, Komagataella is widely used in biochemical research and biotech industries. With strong potential for being an expression system for protein production, as well as being a model organism for genetic study, Komagataella phaffii has become important for biological research and biotech applications. [1] [5]

Taxonomy

According to GBIF: [4]

Komagataella in nature

Natural habitat

In nature, Komagataella is found on trees, such as chestnut trees. [7] They are heterotrophs and they can use several carbon sources for living, like glucose, glycerol and methanol. [8] However, they cannot use lactose.

Reproduction

Komagataella can undergo both asexual reproduction and sexual reproduction, by budding and ascospore. [9] In this case, two types of cells of Komagataella exist: haploid and diploid cells. In the asexual life cycle, haploid cells undergo mitosis for reproduction. In the sexual life cycle, diploid cells undergo sporulation and meiosis. [10] The growth rate of its colonies can vary by a large range, from near to 0 to a doubling time of one hour, which is suitable for industrial processes. [11]

Komagataella as a model organism

In the last few years, Komagataella was investigated and identified as a good model organism with several advantages. First of all, Komagataella can be grown and used easily in lab. Like other widely used yeast models, it has relatively short life span and fast regeneration time. Moreover, some inexpensive culture media have been designed, so that Komagataella can grow quickly on them, with high cell density. [12] Whole genome sequencing for Komagataella had been performed. The K. phaffii GS115 genome has been sequenced by the Flanders Institute for Biotechnology and Ghent University, and published in Nature Biotechnology. [13] The genome sequence and gene annotation can be browsed through the ORCAE system. The complete genomic data allows scientists to identify homologous proteins and evolutionary relationships between other yeast species and Komagataella. In addition, all seven species were sequenced by 2022. [7] Furthermore, Komagataella are single eukaryotic cells, which means researchers could investigate the proteins inside Komagataella. Then the homologous comparison to other more complicated eukaryotic species can be processed, to obtain their functions and origins. [14]

Another advantage of Komagataella is its similarity to the well-studied yeast model — Saccharomyces cerevisiae . As a model organism for biology, S. cerevisiae have been well studied for decades and used by researchers for various purposes throughout history. The two yeast genera; Pichia (sensu lato) and Saccharomyces , have similar growth conditions and tolerances; thus, the culture of Komagataella can be adopted by labs without many modifications. [15] Moreover, unlike S. cerevisiae, Komagataella has the ability to functionally process proteins with large molecular weight, which is useful in a translational host. [16] Considering all the advantages, Komagataella can be usefully employed as both a genetic and experimental model organism.

Komagataella as a genetic model organism

As a genetic model organism, Komagataella can be used for genetic analysis and large-scale genetic crossing, with complete genome data and its ability to carry out complex eukaryotic genetic processing in a relatively small genome. The functional genes for peroxisome assembly were investigated by comparing wild-type and mutant strains of Komagataella. [17]

Komagataella as an experimental model organism

As an experimental model organism, Komagataella was mainly used as the host system for transformation. Due to its abilities of recombination with foreign DNA and processing large proteins, much research has been carried out to investigate the possibility of producing new proteins and the function of artificially designed proteins, using Komagataella as a transformation host. [18] In the last decade, Komagataella was engineered to build expression system platforms, which is a typical application for a standard experimental model organism, as described below.

Komagataella as expression system platform

Komagataella is frequently used as an expression system for the production of heterologous proteins. Several properties make Komagataella suited for this task. Currently, several strains of Komagataella are used for biotechnical purposes, with significant differences among them in growth and protein production. [19] Some common variants possess a mutation in the HIS4 gene, leading to the selection of cells which are transformed successfully with expression vectors. The technology for vector integration into Komagataella genome is similar to that in Saccharomyces cerevisiae. [20]

Advantage

  1. Komagataella is able to grow on simple, inexpensive medium, with high growth rate. Komagataella can grow in either shake flasks or a fermenter, which makes it suitable for both small- and large-scale production. [21]
  2. Komagataella has two alcohol oxidase genes, Aox1 and Aox2, which include strongly inducible promoters. [22] These two genes allow Komagataella to use methanol as a carbon and energy source. The AOX promoters are induced by methanol, and repressed by glucose. Usually, the gene for the desired protein is introduced under the control of the Aox1 promoter, which means that protein production can be induced by the addition of methanol on medium. After several researches, scientists found that the promotor derived from AOX1 gene in Komagataella is extremely suitable to control the expression of foreign genes, which had been transformed into the Komagataella genome, producing heterologous proteins. [23]
  3. With a key trait, Komagataella can grow with extremely high cell density on the culture. This feature is compatible with heterologous protein expression, giving higher yields of production. [24]
  4. The technology required for genetic manipulation of Komagataella is similar to that of Saccharomyces cerevisiae, which is one of the most well-studied yeast model organisms. As a result, the experiment protocol and materials are easy to build for Komagataella. [25]

Disadvantage

As some proteins require chaperonin for proper folding, Komagataella is unable to produce a number of proteins, since it does not contain the appropriate chaperones. The technologies of introducing genes of mammalian chaperonins into the yeast genome and overexpressing existing chaperonins still require improvement. [26] [27]

Comparison with other expression systems

In standard molecular biology research, the bacterium Escherichia coli is the most frequently used organism for expression system, to produce heterologous proteins, due to its features of fast growth rate, high protein production rate, as well as undemanding growth conditions. Protein production in E. coli is usually faster than that in Komagataella, with reasons: Competent E. coli cells can be stored frozen, and thawed before use, whereas Komagataella cells have to be produced immediately before use. Expression yields in Komagataella vary between different clones, so that a large number of clones has to be screened for protein production, to find the best producer. The biggest advantage of Komagataella over E. coli is that Komagataella is capable of forming disulfide bonds and glycosylations in proteins, but E. coli cannot. [28] E. coli might produce a misfolded protein when disulfides are included in final product, leading to inactive or insoluble forms of proteins. [29]

The well-studied Saccharomyces cerevisiae is also used as an expression system with similar advantages over E. coli as Komagataella. However Komagataella has two main advantages over S. cerevisiae in laboratory and industrial settings:

  1. Komagataella, as mentioned above, is a methylotroph, meaning that it can grow with the simple methanol, as the only source of energy — Komagataella can grow fast in cell suspension with reasonably strong methanol solution, which would kill most other micro-organisms. In this case, the expression system is cheap to set up and maintain.
  2. Komagataella can grow up to a very high cell density. Under ideal conditions, it can multiply to the point where the cell suspension is practically a paste. As the protein yield from expression system in a microbe is roughly equal to the product of the proteins produced per cell, which makes Komagataella of great use when trying to produce large quantities of protein without expensive equipment. [28]

Comparing to other expression systems, such as S2-cells from Drosophila melanogaster and Chinese hamster ovary cells, Komagataella usually gives much better yields. Generally, cell lines from multicellular organisms require complex and expensive types of media, including amino acids, vitamins, as well as other growth factors. These types of media significantly increase the cost of producing heterologous proteins. Additionally, Komagataella can grow in media containing only one carbon source and one nitrogen source, which is suitable for isotopic labelling applications, like protein NMR. [28]

Industrial applications

Komagataella have been used in several kinds of biotech industries, such as pharmaceutical industry. All the applications are based on its feature of expressing proteins.

Biotherapeutic production

In the last few years, Komagataella had been used for the production of over 500 types of biotherapeutics, such as IFNγ. At the beginning, one drawback of this protein expression system is the over-glycosylation with high density of mannose structure, which is a potential cause of immunogenicity. [30] [31] In 2006, a research group managed to create a new strain called YSH597. [lower-alpha 1] This strain can express erythropoietin in its normal glycosylation form, by exchanging the enzymes responsible for the fungal type glycosylation, with the mammalian homologs. Thus, the altered glycosylation pattern allowed the protein to be fully functional. [32]

Enzyme production

In food industries, like brewery and bake house, Komagataella is used to produce different kinds of enzymes, as processing aids and food additives, with many functions. For example, some enzymes produced by genetically modified Komagataella can keep the bread soft. Meanwhile, in beer, enzymes could be used to lower the alcohol concentration. [33] Recombinant phospholipase C can degum high-phosphorus oils by breaking down phospholipids. [34]

In animal feed, K. phaffi-produced phytase is used to break down phytic acid, an antinutrient. [34]

Related Research Articles

<span class="mw-page-title-main">Protein production</span>

Protein production is the biotechnological process of generating a specific protein. It is typically achieved by the manipulation of gene expression in an organism such that it expresses large amounts of a recombinant gene. This includes the transcription of the recombinant DNA to messenger RNA (mRNA), the translation of mRNA into polypeptide chains, which are ultimately folded into functional proteins and may be targeted to specific subcellular or extracellular locations.

<span class="mw-page-title-main">Codon usage bias</span> Genetic bias in coding DNA

Codon usage bias refers to differences in the frequency of occurrence of synonymous codons in coding DNA. A codon is a series of three nucleotides that encodes a specific amino acid residue in a polypeptide chain or for the termination of translation.

<span class="mw-page-title-main">Cloning vector</span> Small piece of maintainable DNA

A cloning vector is a small piece of DNA that can be stably maintained in an organism, and into which a foreign DNA fragment can be inserted for cloning purposes. The cloning vector may be DNA taken from a virus, the cell of a higher organism, or it may be the plasmid of a bacterium. The vector contains features that allow for the convenient insertion of a DNA fragment into the vector or its removal from the vector, for example through the presence of restriction sites. The vector and the foreign DNA may be treated with a restriction enzyme that cuts the DNA, and DNA fragments thus generated contain either blunt ends or overhangs known as sticky ends, and vector DNA and foreign DNA with compatible ends can then be joined by molecular ligation. After a DNA fragment has been cloned into a cloning vector, it may be further subcloned into another vector designed for more specific use.

<span class="mw-page-title-main">Expression vector</span> Virus or plasmid designed for gene expression in cells

An expression vector, otherwise known as an expression construct, is usually a plasmid or virus designed for gene expression in cells. The vector is used to introduce a specific gene into a target cell, and can commandeer the cell's mechanism for protein synthesis to produce the protein encoded by the gene. Expression vectors are the basic tools in biotechnology for the production of proteins.

<span class="mw-page-title-main">Recombinant DNA</span> DNA molecules formed by human agency at a molecular level generating novel DNA sequences

Recombinant DNA (rDNA) molecules are DNA molecules formed by laboratory methods of genetic recombination that bring together genetic material from multiple sources, creating sequences that would not otherwise be found in the genome.

The word metagenics uses the prefix meta and the suffix gen. Literally, it means "the creation of something which creates". In the context of biotechnology, metagenics is the practice of engineering organisms to create a specific enzyme, protein, or other biochemicals from simpler starting materials. The genetic engineering of E. coli with the specific task of producing human insulin from starting amino acids is an example. E. coli has also been engineered to digest plant biomass and use it to produce hydrocarbons in order to synthesize biofuels. The applications of metagenics on E. coli also include higher alcohols, fatty-acid based chemicals and terpenes.

Recombineering is a genetic and molecular biology technique based on homologous recombination systems, as opposed to the older/more common method of using restriction enzymes and ligases to combine DNA sequences in a specified order. Recombineering is widely used for bacterial genetics, in the generation of target vectors for making a conditional mouse knockout, and for modifying DNA of any source often contained on a bacterial artificial chromosome (BAC), among other applications.

<i>Pichia</i> Genus of fungi

Pichia is a genus of yeasts in the family Pichiaceae with spherical, elliptical, or oblong acuminate cells. Pichia is a teleomorph, and forms hat-shaped, hemispherical, or round ascospores during sexual reproduction. The anamorphs of some Pichia species are Candida species. The asexual reproduction is by multilateral budding.

Fed-batch culture is, in the broadest sense, defined as an operational technique in biotechnological processes where one or more nutrients (substrates) are fed (supplied) to the bioreactor during cultivation and in which the product(s) remain in the bioreactor until the end of the run. An alternative description of the method is that of a culture in which "a base medium supports initial cell culture and a feed medium is added to prevent nutrient depletion". It is also a type of semi-batch culture. In some cases, all the nutrients are fed into the bioreactor. The advantage of the fed-batch culture is that one can control concentration of fed-substrate in the culture liquid at arbitrarily desired levels.

Cell wall protein 2 (CWP2) is a cell wall protein, produced by Saccharomyces cerevisiae and Saccharomyces pastorianus. It occurs throughout the cell wall and has close homology with the CWP1 gene.

<i>Ogataea polymorpha</i> Species of fungus

Ogataea polymorpha is a methylotrophic yeast with unusual characteristics. It is used as a protein factory for pharmaceuticals.

A yeast expression platform is a strain of yeast used to produce large amounts of proteins, sugars or other compounds for research or industrial uses. While yeast are often more resource-intensive to maintain than bacteria, certain products can only be produced by eukaryotic cells like yeast, necessitating use of a yeast expression platform. Yeasts differ in productivity and with respect to their capabilities to secrete, process and modify proteins. As such, different types of yeast are better suited for different research and industrial applications.

Lachancea kluyveri is an ascomycetous yeast associated with fruit flies, slime fluxes, soils and foods.

<span class="mw-page-title-main">Xylose metabolism</span>

D-Xylose is a five-carbon aldose that can be catabolized or metabolized into useful products by a variety of organisms.

<span class="mw-page-title-main">ATG7</span> Protein-coding gene in the species Homo sapiens

Autophagy related 7 is a protein in humans encoded by ATG7 gene. Related to GSA7; APG7L; APG7-LIKE.

In molecular cloning, a vector is any particle used as a vehicle to artificially carry a foreign nucleic sequence – usually DNA – into another cell, where it can be replicated and/or expressed. A vector containing foreign DNA is termed recombinant DNA. The four major types of vectors are plasmids, viral vectors, cosmids, and artificial chromosomes. Of these, the most commonly used vectors are plasmids. Common to all engineered vectors are an origin of replication, a multicloning site, and a selectable marker.

A subunit vaccine is a vaccine that contains purified parts of the pathogen that are antigenic, or necessary to elicit a protective immune response. Subunit vaccine can be made from dissembled viral particles in cell culture or recombinant DNA expression, in which case it is a recombinant subunit vaccine.

Scheffersomyces stipitis is a species of yeast, belonging to the "CUG Clade" of ascomycetous yeasts. This is a group of fungi that substitute serine for leucine when the CUG codon is encountered. S. stipitis is distantly related to brewer's yeast, Saccharomyces cerevisiae, which uses the conventional codon system. Found, among other places, in the guts of passalid beetles, S. stipitis is capable of both aerobic and oxygen limited fermentation, and has the highest known natural ability of any yeast to directly ferment xylose, converting it to ethanol, a potentially economically valuable trait. Xylose is a hemicellulosic sugar found in all angiosperm plants. As such xylose constitutes the second most abundant carbohydrate moiety in nature. Xylose can be produced from wood or agricultural residues through auto- or acid hydrolysis. Ethanol production from such lignocellulosic residues does not compete with food production through the consumption of grain.

Heterologous expression refers to the expression of a gene or part of a gene in a host organism that does not naturally have the gene or gene fragment in question. Insertion of the gene in the heterologous host is performed by recombinant DNA technology. The purpose of heterologous expression is often to determine the effects of mutations and differential interactions on protein function. It provides an easy path to efficiently express and experiment with combinations of genes and mutants that do not naturally occur.

Aerobic fermentation or aerobic glycolysis is a metabolic process by which cells metabolize sugars via fermentation in the presence of oxygen and occurs through the repression of normal respiratory metabolism. Preference of aerobic fermentation over aerobic respiration is referred to as the Crabtree effect in yeast, and is part of the Warburg effect in tumor cells. While aerobic fermentation does not produce adenosine triphosphate (ATP) in high yield, it allows proliferating cells to convert nutrients such as glucose and glutamine more efficiently into biomass by avoiding unnecessary catabolic oxidation of such nutrients into carbon dioxide, preserving carbon-carbon bonds and promoting anabolism.

References

  1. ↑ YSH597 is based on strain NRRL-Y11430, now considered part of K. phaffi.
  1. 1 2 De Schutter, K., Lin, Y., Tiels, P. (2009). "Genome sequence of the recombinant protein production host Pichia pastoris". Nature Biotechnology . 27 (6): 561–566. doi: 10.1038/nbt.1544 . PMID   19465926.{{cite journal}}: CS1 maint: multiple names: authors list (link)
  2. ↑ Koichi Ogata, Hideo Nishikawa & Masahiro Ohsugi (1969). "A Yeast Capable of Utilizing Methanol". Agricultural and Biological Chemistry . 33 (10): 1519–1520. doi:10.1080/00021369.1969.10859497.
  3. ↑ Yamada, Yuzo; Matsuda, Minako; Maeda, Kojiro; Mikata, Kozaburo (January 1995). "The Phylogenetic Relationships of Methanol-assimilating Yeasts Based on the Partial Sequences of 18S and 26S Ribosomal RNAs: The Proposal of Komagataella Gen. Nov. (Saccharomycetaceae)". Bioscience, Biotechnology, and Biochemistry. 59 (3): 439–444. doi:10.1271/bbb.59.439. PMID   7766181.
  4. 1 2 "Komagataella Y.Yamada, M.Matsuda, K.Maeda & Mikata, 1995". www.gbif.org.
  5. 1 2 Heistinger, Lina; Gasser, Brigitte; Mattanovich, Diethard (2020-07-01). "Microbe Profile: Komagataella phaffii: a methanol devouring biotech yeast formerly known as Pichia pastoris". Microbiology. 166 (7): 614–616. doi: 10.1099/mic.0.000958 . ISSN   1350-0872. PMID   32720891.
  6. ↑ Kurtzman, Cletus Paul (November 2009). "Biotechnological strains of Komagataella (Pichia) pastoris are Komagataella phaffii as determined from multigene sequence analysis". Journal of Industrial Microbiology & Biotechnology. 36 (11): 1435–1438. doi: 10.1007/s10295-009-0638-4 .
  7. 1 2 Heistinger, L; Dohm, JC; Paes, BG; Koizar, D; Troyer, C; Ata, Ö; Steininger-Mairinger, T; Mattanovich, D (25 April 2022). "Genotypic and phenotypic diversity among Komagataella species reveals a hidden pathway for xylose utilization". Microbial Cell Factories. 21 (1): 70. doi: 10.1186/s12934-022-01796-3 . PMC   9036795 . PMID   35468837.
  8. ↑ Rebnegger, C., Vos, T., Graf, A. B., Valli, M., Pronk, J. T., Daran-Lapujade, P., & Mattanovich, D. (2016). "Pichia pastoris exhibits high viability and a low maintenance energy requirement at near-zero specific growth rates". Applied and Environmental Microbiology . 82 (15): 4570–4583. Bibcode:2016ApEnM..82.4570R. doi: 10.1128/AEM.00638-16 . PMC   4984280 . PMID   27208115.{{cite journal}}: CS1 maint: multiple names: authors list (link)
  9. ↑ Kurtzman (1998). "42 - Pichia E.C. Hansen emend. Kurtzman". The Yeasts: A Taxonomic Study. 1: 273–352. doi:10.1016/B978-044481312-1/50046-0. ISBN   9780444813121.
  10. ↑ Zörgö E, Chwialkowska K, Gjuvsland AB, Garré E, Sunnerhagen P, Liti G, Blomberg A, Omholt SW, Warringer J (2013). "Ancient Evolutionary Trade-Offs between Yeast Ploidy States". PLOS Genetics . 9 (3): e1003388. doi: 10.1371/journal.pgen.1003388 . PMC   3605057 . PMID   23555297.{{cite journal}}: CS1 maint: multiple names: authors list (link)
  11. ↑ Kastilan, R., Boes, A., Spiegel, H. (2017). "Improvement of a fermentation process for the production of two PfAMA1-DiCo-based malaria vaccine candidates in Pichia pastoris". Nature . 1 (1): 7. Bibcode:2017NatSR...711991K. doi: 10.1038/s41598-017-11819-4 . PMC   5607246 . PMID   28931852.{{cite journal}}: CS1 maint: multiple names: authors list (link)
  12. ↑ M. M. Guarna G. J. Lesnicki B. M. Tam J. Robinson C. Z. Radziminski D. Hasenwinkle A. Boraston E. Jervis R. T. A. MacGillivray R. F. B. Turner D. G. Kilburn (1997). "On‐line monitoring and control of methanol concentration in shake‐flask cultures of Pichia pastoris". Biotechnology and Bioengineering . 56 (3): 279–286. doi:10.1002/(SICI)1097-0290(19971105)56:3<279::AID-BIT5>3.0.CO;2-G. PMID   18636643.
  13. ↑ De Schutter K, Lin YC, Tiels P, Van Hecke A, Glinka S, Weber-Lehmann J, Rouzé P, Van de Peer Y, Callewaert N (June 2009). "Genome sequence of the recombinant protein production host Pichia pastoris". Nature Biotechnology. 27 (6): 561–6. doi: 10.1038/nbt.1544 . PMID   19465926.
  14. ↑ Brigitte Gasser, Roland Prielhofer, Hans Marx, Michael Maurer, Justyna Nocon, Matthias Steiger, Verena Puxbaum, Michael Sauer & Diethard Mattanovich (2013). "Pichia pastoris: protein production host and model organism for biomedical research". Future Microbiology . 8 (2): 191–208. doi:10.2217/fmb.12.133. PMID   23374125.{{cite journal}}: CS1 maint: multiple names: authors list (link)
  15. ↑ Tran, A., Nguyen, T., Nguyen, C. (2017). "Pichia pastoris versus Saccharomyces cerevisiae: a case study on the recombinant production of human granulocyte-macrophage colony-stimulating factor". BMC Res Notes . 10 (1): 148. doi: 10.1186/s13104-017-2471-6 . PMC   5379694 . PMID   28376863.{{cite journal}}: CS1 maint: multiple names: authors list (link)
  16. ↑ Heidebrecht, Aniela, and Thomas Scheibel (2013). "Recombinant production of spider silk proteins". Advances in Applied Microbiology. 82: 115–153. doi:10.1016/B978-0-12-407679-2.00004-1. ISBN   9780124076792. PMID   23415154.{{cite journal}}: CS1 maint: multiple names: authors list (link)
  17. ↑ Gould, S. J., McCollum, D., Spong, A. P., Heyman, J. A., & Subramani, S. (1992). "Development of the yeast Pichia pastoris as a model organism for a genetic and molecular analysis of peroxisome assembly". The Yeasts: A Taxonomic Study. 8 (8): 613–628. doi:10.1002/yea.320080805. PMID   1441741. S2CID   8840145.{{cite journal}}: CS1 maint: multiple names: authors list (link)
  18. ↑ Cregg, J. M., Barringer, K. J., Hessler, A. Y., & Madden, K. R. (1985). "Pichia pastoris as a host system for transformations". Molecular and Cellular Biology . 5 (12): 3376–3385. doi:10.1128/MCB.5.12.3376. PMC   369166 . PMID   3915774.{{cite journal}}: CS1 maint: multiple names: authors list (link)
  19. ↑ Brady, J.R. (2020). "Comparative genome-scale analysis of Pichia pastoris variants informs selection of an optimal base strain". Biotechnology and Bioengineering. 117 (2): 543–555. doi:10.1002/bit.27209. PMC   7003935 . PMID   31654411.
  20. ↑ Higgins, D. R., & Cregg, J. M. (1998). "Introduction to Pichia pastoris". Pichia Protocols. Methods in Molecular Biology. Vol. 103. pp. 1–15. doi:10.1385/0-89603-421-6:1. ISBN   0-89603-421-6. PMID   9680629.{{cite book}}: CS1 maint: multiple names: authors list (link)
  21. ↑ Wenhui Zhang Mark A. Bevins Bradley A. Plantz Leonard A. Smith Michael M. Meagher. (2000). "Modeling Pichia pastoris growth on methanol and optimizing the production of a recombinant protein, the heavy‐chain fragment C of botulinum neurotoxin, serotype A". Biotechnology and Bioengineering . 70 (1): 1–8. doi:10.1002/1097-0290(20001005)70:1<1::AID-BIT1>3.0.CO;2-Y. PMID   10940857.
  22. ↑ Daly R, Hearn MT (2005). "Expression of heterologous proteins in Pichia pastoris: a useful experimental tool in protein engineering and production". Journal of Molecular Recognition. 18 (2): 119–38. doi:10.1002/jmr.687. PMID   15565717. S2CID   7476149.
  23. ↑ Romanos, Mike. (1995). "Advances in the use of Pichia pastoris for high-level gene expression". Current Opinion in Biotechnology . 6 (5): 527–533. doi:10.1016/0958-1669(95)80087-5.
  24. ↑ Zhou, X., Yu, Y., Tao, J., & Yu, L. (2014). "Production of LYZL6, a novel human c-type lysozyme, in recombinant Pichia pastoris employing high cell density fed-batch fermentation". Journal of Bioscience and Bioengineering . 118 (4): 420–425. doi:10.1016/j.jbiosc.2014.03.009. PMID   24745549.{{cite journal}}: CS1 maint: multiple names: authors list (link)
  25. ↑ Morton, C. L., & Potter, P. M. (2000). "Comparison of Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris, Spodoptera frugiperda, and COS7 cells for recombinant gene expression". Molecular Biotechnology . 16 (3): 193–202. doi:10.1385/MB:16:3:193. PMID   11252804. S2CID   22792748.{{cite journal}}: CS1 maint: multiple names: authors list (link)
  26. ↑ Bankefa, OE; Wang, M; Zhu, T; Li, Y (July 2018). "Hac1p homologues from higher eukaryotes can improve the secretion of heterologous proteins in the yeast Pichia pastoris". Biotechnology Letters. 40 (7): 1149–1156. doi: 10.1007/s10529-018-2571-y . PMID   29785668. S2CID   29155989.
  27. ↑ Yu, Xiao-Wei; Sun, Wei-Hong; Wang, Ying-Zheng; Xu, Yan (24 November 2017). "Identification of novel factors enhancing recombinant protein production in multi-copy Komagataella phaffii based on transcriptomic analysis of overexpression effects". Scientific Reports. 7 (1): 16249. Bibcode:2017NatSR...716249Y. doi: 10.1038/s41598-017-16577-x . PMC   5701153 . PMID   29176680.
  28. 1 2 3 Cregg JM, Tolstorukov I, Kusari A, Sunga J, Madden K, Chappell T (2009). "Chapter 13 Expression in the Yeast Pichia pastoris". Guide to Protein Purification, 2nd Edition. Methods in Enzymology. Vol. 463. pp. 169–89. doi:10.1016/S0076-6879(09)63013-5. ISBN   978-0-12-374536-1. PMID   19892173.{{cite book}}: |journal= ignored (help)
  29. ↑ Brondyk WH (2009). "Chapter 11 Selecting an Appropriate Method for Expressing a Recombinant Protein". Guide to Protein Purification, 2nd Edition. Methods in Enzymology. Vol. 463. pp. 131–47. doi:10.1016/S0076-6879(09)63011-1. ISBN   978-0-12-374536-1. PMID   19892171.{{cite book}}: |journal= ignored (help)
  30. ↑ Razaghi A, Tan E, Lua LH, Owens L, Karthikeyan OP, Heimann K (January 2017). "Is Pichia pastoris a realistic platform for industrial production of recombinant human interferon gamma?". Biologicals. 45: 52–60. doi:10.1016/j.biologicals.2016.09.015. PMID   27810255. S2CID   28204059.
  31. ↑ Ali Razaghi; Roger Huerlimann; Leigh Owens; Kirsten Heimann (2015). "Increased expression and secretion of recombinant hIFNγ through amino acid starvation-induced selective pressure on the adjacent HIS4 gene in Pichia pastoris" (PDF). European Pharmaceutical Journal. 62 (2): 43–50. doi: 10.1515/afpuc-2015-0031 .
  32. ↑ Hamilton SR, Davidson RC, Sethuraman N, Nett JH, Jiang Y, Rios S, Bobrowicz P, Stadheim TA, Li H, Choi BK, Hopkins D, Wischnewski H, Roser J, Mitchell T, Strawbridge RR, Hoopes J, Wildt S, Gerngross TU (September 2006). "Humanization of yeast to produce complex terminally sialylated glycoproteins". Science. 313 (5792): 1441–3. Bibcode:2006Sci...313.1441H. doi:10.1126/science.1130256. PMID   16960007. S2CID   43334198.
  33. ↑ Spohner, S. C., Müller, H., Quitmann, H., & Czermak, P. (2015). "Expression of enzymes for the usage in food and feed industry with Pichia pastoris". Journal of Biotechnology. 202: 420–425. doi:10.1016/j.jbiotec.2015.01.027. PMID   25687104.{{cite journal}}: CS1 maint: multiple names: authors list (link)
  34. 1 2 Barone, GD; Emmerstorfer-Augustin, A; Biundo, A; Pisano, I; Coccetti, P; Mapelli, V; Camattari, A (26 February 2023). "Industrial Production of Proteins with Pichia pastoris-Komagataella phaffii". Biomolecules. 13 (3): 441. doi: 10.3390/biom13030441 . PMC   10046876 . PMID   36979376.
This page is based on this Wikipedia article
Text is available under the CC BY-SA 4.0 license; additional terms may apply.
Images, videos and audio are available under their respective licenses.