Purification and immunodetection of the complete recombinant HER-2[neu] receptor produced in yeast

Chamas, Alexandre; Giersberg, Martin; Friedrich, Katrin; Sonntag, Frank; Kunze, D.; Uhlig, Steffen; Simon, Kirsten; Baronian, Keith; Kunze, Gotthard

doi:10.1016/j.pep.2014.10.004

Cited by 5 publications

(4 citation statements)

References 35 publications

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“…The non-conventional yeast Arxula adeninivorans , previously described as a valuable platform for production of recombinant proteins (Rauter et al 2014; Chamas et al 2015; Jankowska et al 2013) and as a producer of the copolymer PHB-V (Terentiev et al 2004), was proposed for the synthesis of (R)-3-HB. The advantages of using A. adeninivorans as a host are its wide substrate spectrum, its robustness and non-pathogenicity (Lindenkamp et al 2012).…”

Section: Introductionmentioning

confidence: 99%

Production of (R)-3-hydroxybutyric acid by Arxula adeninivorans

Biernacki

Riechen²,

Hähnel

et al. 2017

AMB Expr

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(R)-3-hydroxybutyric acid can be used in industrial and health applications. The synthesis pathway comprises two enzymes, β-ketothiolase and acetoacetyl-CoA reductase which convert cytoplasmic acetyl-CoA to (R)-3-hydroxybutyric acid [(R)-3-HB] which is released into the culture medium. In the present study we used the non-conventional yeast, Arxula adeninivorans, for the synthesis enantiopure (R)-3-HB. To establish optimal production, we investigated three different endogenous yeast thiolases (Akat1p, Akat2p, Akat4p) and three bacterial thiolases (atoBp, thlp, phaAp) in combination with an enantiospecific reductase (phaBp) from Cupriavidus necator H16 and endogenous yeast reductases (Atpk2p, Afox2p). We found that Arxula is able to release (R)-3-HB used an existing secretion system negating the need to engineer membrane transport. Overexpression of thl and phaB genes in organisms cultured in a shaking flask resulted in 4.84 g L−1 (R)-3-HB, at a rate of 0.023 g L−1 h−1 over 214 h. Fed-batch culturing with glucose as a carbon source did not improve the yield, but a similar level was reached with a shorter incubation period [3.78 g L−1 of (R)-3-HB at 89 h] and the rate of production was doubled to 0.043 g L−1 h−1 which is higher than any levels in yeast reported to date. The secreted (R)-3-HB was 99.9% pure. This is the first evidence of enantiopure (R)-3-HB synthesis using yeast as a production host and glucose as a carbon source.

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Section: Introductionmentioning

confidence: 99%

Production of (R)-3-hydroxybutyric acid by Arxula adeninivorans

Biernacki

Riechen²,

Hähnel

et al. 2017

AMB Expr

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“…Furthermore, an HER-2 cancer cell detector utilizing surface plasmon resonance (SPR) has been constructed for the rapid diagnosis of a particularly aggressive type of breast cancer (Chamas et al, 2015b).…”

Section: Arxula Adeninivorans As a Biocomponent For Biosensorsmentioning

confidence: 99%

Blastobotrys (Arxula) adeninivorans: a promising alternative yeast for biotechnology and basic research

Malak

Baronian

Kunze

2016

Yeast

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Blastobotrys adeninivorans (syn. Arxula adeninivorans) is a non-conventional, non-pathogenic, imperfect, haploid yeast, belonging to the subphylum Saccharomycotina, which has to date received comparatively little attention from researchers. It possesses unusual properties such as thermo- and osmotolerance, and a broad substrate spectrum. Depending on the cultivation temperature B. (A.) adeninivorans exhibits different morphological forms and various post-translational modifications and protein expression properties that are strongly correlated with the morphology. The genome has been completely sequenced and, in addition, there is a well-developed transformation/expression platform, which makes rapid, simple gene manipulations possible. This yeast species is a very good host for homologous and heterologous gene expression and is also a useful gene donor. Blastobotrys (A.) adeninivorans is able to use a very wide range of substrates as carbon and/or nitrogen sources and is an interesting organism owing to the presence of many metabolic pathways, for example degradation of n-butanol, purines and tannin. In addition, its unusual properties and robustness make it a useful bio-component for whole cell biosensors. There are currently a number of products on the market produced by B. (A.) adeninivorans and further investigation may contribute further innovative solutions for current challenges that exist in the biotechnology industry. Additionally it may become a useful alternative to existing commercial yeast strains and as a model organism in research. In this review we present information relevant to the exploitation of B. (A.) adeninivorans in research and industrial settings. Copyright © 2016 John Wiley & Sons, Ltd.

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“…The optical system approach is derived from a system previously developed for parallel SPR analysis on polymer chips 12 the performance of which has been demonstrated recently 20,21 . A simplified overview of the optical system modified for BSW based analysis is presented in Figure 4.…”

Section: Optical Analysismentioning

confidence: 99%

Biosensing platform combining label-free and labelled analysis using Bloch surface waves

et al. 2015

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Bloch surface waves (BSW) propagating at the boundary of truncated photonic crystals (1D-PC) have emerged as an attractive approach for label-free sensing in plasmon-like sensor configurations. Due to the very low losses in such dielectric thin film stacks, BSW feature very low angular resonance widths compared to the surface plasmon resonance (SPR) case. Besides label-free operation, the large field enhancement and the absence of quenching allow utilizing BSW coupled fluorescence detection to additionally sense the presence of fluorescent labels. This approach can be adapted to the case of angularly resolved resonance detection, thus giving rise to a combined label-free / labelled biosensor platform. It features a parallel analysis of multiple spots arranged as a one-dimensional array inside a microfluidic channel of a disposable chip. Application of such a combined biosensing approach to the detection of the Angiopoietin-2 cancer biomarker in buffer solutions is reported.

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Purification and immunodetection of the complete recombinant HER-2[neu] receptor produced in yeast

Cited by 5 publications

References 35 publications

Production of (R)-3-hydroxybutyric acid by Arxula adeninivorans

Production of (R)-3-hydroxybutyric acid by Arxula adeninivorans

Blastobotrys (Arxula) adeninivorans: a promising alternative yeast for biotechnology and basic research

Biosensing platform combining label-free and labelled analysis using Bloch surface waves

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