Yeast expression platforms

Böer, Erik; Steinborn, Gerhard; Kunze, Gotthard; Gellissen, Gerd

doi:10.1007/s00253-007-1209-0

Cited by 93 publications

(56 citation statements)

References 83 publications

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“…The heterologous production of EntA by bacterial hosts has been attained through the expression of its native biosynthetic genes (29), by exchange or replacement of the EntA leader peptide and/or dedicated processing and secretion systems (33), or by fusion of mature EntA to signal peptides that act as secretion signals (10,28,32). Recently, several yeast platforms have been developed for the large-scale expression of proteins (5,7,13,19). However, the heterologous production of bacteriocins by yeasts has not yet been fully exploited.…”

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confidence: 99%

Cloning, Production, and Functional Expression of the Bacteriocin Enterocin A, Produced by Enterococcus faecium T136, by the Yeasts Pichia pastoris, Kluyveromyces lactis, Hansenula polymorpha, and Arxula adeninivorans

Borrero

Kunze

Jiménez

et al. 2012

Appl Environ Microbiol

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bThe bacteriocin enterocin A (EntA) produced by Enterococcus faecium T136 has been successfully cloned and produced by the yeasts Pichia pastoris X-33EA, Kluyveromyces lactis GG799EA, Hansenula polymorpha KL8-1EA, and Arxula adeninivorans G1212EA. Moreover, P. pastoris X-33EA and K. lactis GG799EA produced EntA in larger amounts and with higher antimicrobial and specific antimicrobial activities than the EntA produced by E. faecium T136.

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confidence: 99%

Cloning, Production, and Functional Expression of the Bacteriocin Enterocin A, Produced by Enterococcus faecium T136, by the Yeasts Pichia pastoris, Kluyveromyces lactis, Hansenula polymorpha, and Arxula adeninivorans

Borrero

Kunze

Jiménez

et al. 2012

Appl Environ Microbiol

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“…These issues are generally overcome by using alternative recombinant protein expression systems, such as P. pastoris (Macauley-Patrick et al, 2005;Sreekrishna et al, 1988;Eckart and Bussineau, 1996;Jahic et al, 2006;Hamilton and Gerngross, 2007). To date, P. pastoris has been successfully used for the expression of more than 420 heterologous recombinant proteins (MacauleyPatrick et al, 2005;Lin Cereghino et al, 2008;Boer et al, 2007). Green fluorescent protein (GFP) from the jellyfish Aequorea victoria has become one of the most widely used reporter proteins in recent years.…”

Section: Discussionmentioning

confidence: 99%

Expression of GFP using Pichia pastoris vectors with zeocin or G‐418 sulphate as the primary selectable marker

Papakonstantinou

Harris

Hearn

2009

Yeast

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Pichia pastoris is a popular host organism for expressing heterologous proteins, and various expression vectors for this yeast are currently available. Recently, vectors containing novel dominant antibiotic resistance markers have become a strong and developing field of research for this methylotropic yeast strain. We have developed new P. pastoris expression vectors, the pPICKanMX6 and pPICKanMX6α series. These vectors were constructed by replacing the zeocin resistance gene of the pPICZA, B, C and pPICZαA, B and C vectors with the Tn903 kan R marker from pFA6a KanMX6, which confers G-418 sulphate resistance in P. pastoris. The limits of antibiotic resistance in two transformant yeast strains were investigated, and the selection marker was shown to be stably retained. To demonstrate their usefulness, a gene encoding hexa-histidine-tagged green fluorescent protein (GFPH6) was cloned into one of the new vectors and GFP expression examined in P. pastoris cells. The protein expression levels using the pPICKanMX6B vector were comparable with that using the original plasmid, based on zeocin resistance as seen by yeast cell fluorescence. Moreover, GFPH6 was able to be isolated by immobilized metal ion affinity chromatography (IMAC) from lysates of both yeast strains. A model reporter construct has been used to demonstrate successful recombinant protein expression and its subsequent purification using these new vectors. Corresponding vectors can now also be engineered with foreign gene expression under the control of various different promoters, to increase the flexibility of P. pastoris as a cellular factory for heterologous protein production.

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“…Apart from S. cerevisiae , a number of alternative yeast expression systems have been developed (reviewed in [ 103 ]). Among them, the methylotrophic yeast Pichia pastoris as a cellular host for the expression of recombinant proteins has become increasing popular in recent times.…”

Section: Insect Cellsmentioning

confidence: 99%

General Introduction: Recombinant Protein Production and Purification of Insoluble Proteins

Ferrer-Miralles

Saccardo

Corchero

et al. 2014

Methods in Molecular Biology

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Proteins are synthesized in heterologous systems because of the impossibility to obtain satisfactory yields from natural sources. The production of soluble and functional recombinant proteins is among the main goals in the biotechnological field. In this context, it is important to point out that under stress conditions, protein folding machinery is saturated and this promotes protein misfolding and, consequently, protein aggregation. Thus, the selection of the optimal expression organism and the most appropriate growth conditions to minimize the formation of insoluble proteins should be done according to the protein characteristics and downstream requirements. Escherichia coli is the most popular recombinant protein expression system despite the great development achieved so far by eukaryotic expression systems. Besides, other prokaryotic expression systems, such as lactic acid bacteria and psychrophilic bacteria, are gaining interest in this field. However, it is worth mentioning that prokaryotic expression system poses, in many cases, severe restrictions for a successful heterologous protein production. Thus, eukaryotic systems such as mammalian cells, insect cells, yeast, filamentous fungus, and microalgae are an interesting alternative for the production of these difficult-to-express proteins.

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Yeast expression platforms

Cited by 93 publications

References 83 publications

Cloning, Production, and Functional Expression of the Bacteriocin Enterocin A, Produced by Enterococcus faecium T136, by the Yeasts Pichia pastoris, Kluyveromyces lactis, Hansenula polymorpha, and Arxula adeninivorans

Cloning, Production, and Functional Expression of the Bacteriocin Enterocin A, Produced by Enterococcus faecium T136, by the Yeasts Pichia pastoris, Kluyveromyces lactis, Hansenula polymorpha, and Arxula adeninivorans

Expression of GFP using Pichia pastoris vectors with zeocin or G‐418 sulphate as the primary selectable marker

General Introduction: Recombinant Protein Production and Purification of Insoluble Proteins

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