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Kluyveromyces marxianus as a host for heterologous protein synthesis.
Appl Microbiol Biotechnol. 2016 Jul; 100(14):6193-6208.AM

Abstract

The preferentially respiring and thermotolerant yeast Kluyveromyces marxianus is an emerging host for heterologous protein synthesis, surpassing the traditional preferentially fermenting yeast Saccharomyces cerevisiae in some important aspects: K . marxianus can grow at temperatures 10 °C higher than S. cerevisiae, which may result in decreased costs for cooling bioreactors and reduced contamination risk; has ability to metabolize a wider variety of sugars, such as lactose and xylose; is the fastest growing eukaryote described so far; and does not require special cultivation techniques (such as fed-batch) to avoid fermentative metabolism. All these advantages exist together with a high secretory capacity, performance of eukaryotic post-translational modifications, and with a generally regarded as safe (GRAS) status. In the last years, replication origins from several Kluyveromyces spp. have been used for the construction of episomal vectors, and also integrative strategies have been developed based on the tendency for non-homologous recombination displayed by K. marxianus. The recessive URA3 auxotrophic marker and the dominant Kan(R) are mostly used for selection of transformed cells, but other markers have been made available. Homologous and heterologous promoters and secretion signals have been characterized, with the K. marxianus INU1 expression and secretion system being of remarkable functionality. The efficient synthesis of roughly 50 heterologous proteins has been demonstrated, including one thermophilic enzyme. In this mini-review, we summarize the physiological characteristics of K. marxianus relevant for its use in the efficient synthesis of heterologous proteins, the efforts performed hitherto in the development of a molecular toolbox for this purpose, and some successful examples.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Gombert AK
School of Food Engineering, University of Campinas, Rua Monteiro Lobato, 80, Campinas, SP, 13083-862, Brazil.
Madeira JV
School of Food Engineering, University of Campinas, Rua Monteiro Lobato, 80, Campinas, SP, 13083-862, Brazil.
Cerdán ME
Grupo EXPRELA, Centro de Investigacións Científicas Avanzadas (CICA), Departamento de Bioloxía Celular e Molecular, Facultade de Ciencias, Universidade da Coruña, Campus de A Coruña, 15071, A Coruña, Spain.
González-Siso MI
Grupo EXPRELA, Centro de Investigacións Científicas Avanzadas (CICA), Departamento de Bioloxía Celular e Molecular, Facultade de Ciencias, Universidade da Coruña, Campus de A Coruña, 15071, A Coruña, Spain. migs@udc.es.

MeSH

FermentationFungal ProteinsGene Expression Regulation, FungalGenomicsIndustrial MicrobiologyKluyveromycesLactosePlasmidsPromoter Regions, GeneticProtein BiosynthesisProtein Processing, Post-TranslationalSaccharomyces cerevisiaeTemperatureXylose

Pub Type(s)

Journal Article
Review

Language

eng

PubMed ID

27260286

Citation

Gombert, Andreas K., et al. "Kluyveromyces Marxianus as a Host for Heterologous Protein Synthesis." Applied Microbiology and Biotechnology, vol. 100, no. 14, 2016, pp. 6193-6208.
Gombert AK, Madeira JV, Cerdán ME, et al. Kluyveromyces marxianus as a host for heterologous protein synthesis. Appl Microbiol Biotechnol. 2016;100(14):6193-6208.
Gombert, A. K., Madeira, J. V., Cerdán, M. E., & González-Siso, M. I. (2016). Kluyveromyces marxianus as a host for heterologous protein synthesis. Applied Microbiology and Biotechnology, 100(14), 6193-6208. https://doi.org/10.1007/s00253-016-7645-y
Gombert AK, et al. Kluyveromyces Marxianus as a Host for Heterologous Protein Synthesis. Appl Microbiol Biotechnol. 2016;100(14):6193-6208. PubMed PMID: 27260286.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Kluyveromyces marxianus as a host for heterologous protein synthesis. AU - Gombert,Andreas K, AU - Madeira,José Valdo,Jr AU - Cerdán,María-Esperanza, AU - González-Siso,María-Isabel, Y1 - 2016/06/03/ PY - 2016/02/22/received PY - 2016/05/25/accepted PY - 2016/05/22/revised PY - 2016/6/5/entrez PY - 2016/6/5/pubmed PY - 2017/3/9/medline KW - Cell factory KW - Eukaryotic post-translational modifications KW - Heterologous protein secretion KW - Heterologous protein synthesis KW - Kluyveromyces marxianus KW - Thermotolerant yeast SP - 6193 EP - 6208 JF - Applied microbiology and biotechnology JO - Appl Microbiol Biotechnol VL - 100 IS - 14 N2 - The preferentially respiring and thermotolerant yeast Kluyveromyces marxianus is an emerging host for heterologous protein synthesis, surpassing the traditional preferentially fermenting yeast Saccharomyces cerevisiae in some important aspects: K . marxianus can grow at temperatures 10 °C higher than S. cerevisiae, which may result in decreased costs for cooling bioreactors and reduced contamination risk; has ability to metabolize a wider variety of sugars, such as lactose and xylose; is the fastest growing eukaryote described so far; and does not require special cultivation techniques (such as fed-batch) to avoid fermentative metabolism. All these advantages exist together with a high secretory capacity, performance of eukaryotic post-translational modifications, and with a generally regarded as safe (GRAS) status. In the last years, replication origins from several Kluyveromyces spp. have been used for the construction of episomal vectors, and also integrative strategies have been developed based on the tendency for non-homologous recombination displayed by K. marxianus. The recessive URA3 auxotrophic marker and the dominant Kan(R) are mostly used for selection of transformed cells, but other markers have been made available. Homologous and heterologous promoters and secretion signals have been characterized, with the K. marxianus INU1 expression and secretion system being of remarkable functionality. The efficient synthesis of roughly 50 heterologous proteins has been demonstrated, including one thermophilic enzyme. In this mini-review, we summarize the physiological characteristics of K. marxianus relevant for its use in the efficient synthesis of heterologous proteins, the efforts performed hitherto in the development of a molecular toolbox for this purpose, and some successful examples. SN - 1432-0614 UR - https://www.unboundmedicine.com/medline/citation/27260286/Kluyveromyces_marxianus_as_a_host_for_heterologous_protein_synthesis_ DB - PRIME DP - Unbound Medicine ER -