Comparison of expression systems in the yeastsSaccharomyces cerevisiae, Hansenula polymorpha, Klyveromyces lactis, Schizosaccharomyces pombe andYarrowia lipolytica. Cloning of two novel promoters fromYarrowia lipolytica

Müller, Sven; Sandal, Thomas; Kamp‐Hansen, Peter; Dalbøge, Henrik

doi:10.1002/(sici)1097-0061(1998100)14:14<1267::aid-yea327>3.0.co;2-2

Cited by 208 publications

(29 citation statements)

References 41 publications

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“…Y. lipolytica is a recognized system for heterologous protein expression (Barth and Gaillardin 1996; Madzak et al 2004). Direct comparison of different expression platforms— Y. lipolytica , S. cerevisiae , P. pastoris , Arxula adeninivorans , Hansenula polymorpha , Kluyveromyces lactis , and Schizosaccharomyces pombe (Gellissen et al 2005; Müller et al 1998)—demonstrated that Y. lipolytica is characterized by several advantageous traits for heterologous protein expression over the other expression systems. Several detailed review papers regarding the applied strategies used vectors, and heterologous proteins expressed in Y. lipolytica are available in the literature (e.g., Madzak et al 2004; Madzak and Beckerich 2013).…”

Section: Introductionmentioning

confidence: 99%

Cloning, expression, and purification of insect (Sitophilus oryzae) alpha-amylase, able to digest granular starch, in Yarrowia lipolytica host

Celińska

Białas

Borkowska

et al. 2014

Appl Microbiol Biotechnol

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Raw-starch-digesting enzymes (RSDE) are of major importance for industrial applications, as their usage greatly simplifies the starch processing pipeline. To date, only microbial RSDE have gained considerable attention, since only microbial production of enzymes meets industrial demands. In this study, α-amylase from rice weevil (Sitophilus oryzae), the major rice pest, was cloned and expressed in Yarrowia lipolytica Po1g strain. The enzyme was secreted into the culture medium, and the peak activity (81 AU/L) was reached after only 29 h of culturing in 5-L bioreactors. Through simple purification procedure of ammonium sulfate precipitation and affinity chromatography, it was possible to purify the enzyme to apparent homogeneity (25-fold purification factor, at 5 % yield). The optimal conditions for the α-amylase activity were pH 5.0 and a temperature of 40 °C. The α-amylase studied here did not show any obligate requirement for Ca2+ ions. The recombinant α-amylase appeared to efficiently digest granular starch from pea, amaranth, waxy corn, and waxy rice.

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Section: Introductionmentioning

confidence: 99%

Cloning, expression, and purification of insect (Sitophilus oryzae) alpha-amylase, able to digest granular starch, in Yarrowia lipolytica host

Celińska

Białas

Borkowska

et al. 2014

Appl Microbiol Biotechnol

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“…The most attractive yeast, especially in terms of performance reproducibility, was Y. lipolytica [3]. …”

Section: Introductionmentioning

confidence: 99%

“…Y. lipolytica was also distinguished for its ability to secrete naturally several metabolites and proteins in large amounts into the culture medium. But, probably the most important characteristic of Y. lipolytica as a host is the convenience of its secretory apparatus; high efficiency, co-translational pathway and low overglycosylation, in some regards closer to that of mammalian cells than to those of many yeasts [3,4,6]. Moreover, development of molecular and cellular tools and a wide range of host strains make Y. lipolytica one of the most attractive host for the expression of heterologous proteins [6].…”

Section: Introductionmentioning

confidence: 99%

Development of a cultivation process for the enhancement of human interferon alpha 2b production in the oleaginous yeast, Yarrowia lipolytica

et al. 2011

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BackgroundAs an oleaginous yeast, Yarrowia lipolytica is able to assimilate hydrophobic substrates. This led to the isolation of several promoters of key enzymes of this catabolic pathway. Less is known about the behavior of Y. lipolytica in large bioreactors using these substrates. There is therefore a lack of established know-how concerning high cell density culture protocols of this yeast. Consequently, the establishment of suitable induction conditions is required, to maximize recombinant protein production under the control of these promoters.ResultsHuman interferon α2b (huIFN α2b) production in Yarrowia lipolytica was used as a model for the enhancement of recombinant protein production under the control of the oleic acid (OA)-inducible promoter POX2. Cell viability and heterologous protein production were enhanced by exponential glucose feeding, to generate biomass before OA induction. The optimal biomass level before induction was determined (73 g L-1), and glucose was added with oleic acid during the induction phase. Several oleic acid feeding strategies were assessed. Continuous feeding with OA at a ratio of 0.02 g OA per g dry cell weight increased huIFNα2b production by a factor of 1.88 (425 mg L-1) and decreased the induction time (by a factor of 2.6, 21 h). huIFN α2b degradation by an aspartic protease secreted by Y. lipolytica was prevented by adding pepstatin (10 μM), leading to produce a 19-fold more active huIFN α2b (26.2 × 107 IU mg-1).ConclusionY. lipolytica, a generally regarded as safe (GRAS) microorganism is one of the most promising non conventional yeasts for the production of biologically active therapeutic proteins under the control of hydrophobic substrate-inducible promoter.

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“…It is thus important to have a range of possible hosts to investigate the best one for efficient scFv production. Yarrowia lipolytica and Kluyveromyces lactis, two non-conventional yeasts, can efficiently secrete heterologous proteins (Gellissen & Hollenberg, 1997 ;van der Berg et al, 1990 ;Fleer, 1992 ;Muller et al, 1998 ;Dominguez et al, 1998). Both are generally recognized as safe (GRAS) organisms.…”

Section: Introductionmentioning

confidence: 99%

“…Y. lipolytica is able to secrete approximately 1 g alkaline extracellular protease (AEP) l −" into the medium under optimal conditions (Barth & Gaillardin, 1997), suggesting good potential for secretion of heterologous proteins. The promoter of the XPR2 gene encoding AEP has been used to direct heterologous protein expression in Y. lipolytica (for examples see Park et al, 1997 ;Muller et al, 1998). Regulation of this promoter is very complex thus limiting its industrial use.…”

Section: Introductionmentioning

confidence: 99%

Secretion of active anti-Ras single-chain Fv antibody by the yeasts Yarrowia lipolytica and Kluyveromyces lactis

Swennen

Paul²,

Vernis

et al. 2002

Microbiology

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Comparison of expression systems in the yeastsSaccharomyces cerevisiae, Hansenula polymorpha, Klyveromyces lactis, Schizosaccharomyces pombe andYarrowia lipolytica. Cloning of two novel promoters fromYarrowia lipolytica

Cited by 208 publications

References 41 publications

Cloning, expression, and purification of insect (Sitophilus oryzae) alpha-amylase, able to digest granular starch, in Yarrowia lipolytica host

Cloning, expression, and purification of insect (Sitophilus oryzae) alpha-amylase, able to digest granular starch, in Yarrowia lipolytica host

Development of a cultivation process for the enhancement of human interferon alpha 2b production in the oleaginous yeast, Yarrowia lipolytica

Secretion of active anti-Ras single-chain Fv antibody by the yeasts Yarrowia lipolytica and Kluyveromyces lactis

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