53 research outputs found
Fermentation of deproteinized cheese whey powder solutions to ethanol by engineered Saccharomyces cerevisiae : effect of supplementation with corn steep liquor and repeated-batch operation with biomass recycling by flocculation
The lactose in cheese whey is an interesting
substrate for the production of bulk commodities such as
bio-ethanol, due to the large amounts of whey surplus
generated globally. In this work, we studied the performance
of a recombinant Saccharomyces cerevisiae strain
expressing the lactose permease and intracellular ß-galactosidase
from Kluyveromyces lactis in fermentations of
deproteinized concentrated cheese whey powder solutions.
Supplementation with 10 g/l of corn steep liquor significantly
enhanced whey fermentation, resulting in the production
of 7.4% (v/v) ethanol from 150 g/l initial lactose in
shake-flask fermentations, with a corresponding productivity
of 1.2 g/l/h. The flocculation capacity of the yeast
strain enabled stable operation of a repeated-batch process
in a 5.5-l air-lift bioreactor, with simple biomass recycling
by sedimentation of the yeast flocs. During five consecutive
batches, the average ethanol productivity was 0.65 g/l/h
and ethanol accumulated up to 8% (v/v) with lactose-toethanol
conversion yields over 80% of theoretical. Yeast
viability (>97%) and plasmid retention (>84%) remained
high throughout the operation, demonstrating the stability
and robustness of the strain. In addition, the easy and
inexpensive recycle of the yeast biomass for repeated utilization
makes this process economically attractive for
industrial implementation.Fundação para a Ciência e a Tecnologia (FCT)LACTOGAL-Produtos Alimentares S.A.Companhia Portuguesa de Amidos, S.A
TGA2 signaling in response to reactive electrophile species is not dependent on cysteine modification of TGA2
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.Reactive electrophile species (RES), including prostaglandins, phytoprostanes and 12-oxo phytodienoic acid (OPDA), activate detoxification responses in plants and animals. However, the pathways leading to the activation of defense reactions related to abiotic or biotic stress as a function of RES formation, accumulation or treatment are poorly understood in plants. Here, the thiol-modification of proteins, including the RES-activated basic region/leucine zipper transcription factor TGA2, was studied. TGA2 contains a single cysteine residue (Cys186) that was covalently modified by reactive cyclopentenones but not required for induction of detoxification genes in response to OPDA or prostaglandin A1. Activation of the glutathione-S-transferase 6 (GST6) promoter was responsive to cyclopentenones but not to unreactive cyclopentanones, including jasmonic acid suggesting that thiol reactivity of RES is important to activate the TGA2-dependent signaling pathway resulting in GST6 activation We show that RES modify thiols in numerous proteins in vivo, however, thiol reactivity alone appears not to be sufficient for biological activity as demonstrated by the failure of several membrane permeable thiol reactive reagents to activate the GST6 promoter.Peer reviewedFinal Published versio
VEGFR2 Translocates to the Nucleus to Regulate Its Own Transcription
Vascular Endothelial Growth Factor Receptor-2 (VEGFR2) is the major mediator of the angiogenic effects of VEGF. In addition to its well known role as a membrane receptor that activates multiple signaling pathways, VEGFR2 also has a nuclear localization. However, what VEGFR2 does in the nucleus is still unknown. In the present report we show that, in endothelial cells, nuclear VEGFR2 interacts with several nuclear proteins, including the Sp1, a transcription factor that has been implicated in the regulation of genes needed for angiogenesis. By in vivo chromatin immunoprecipitation (ChIP) assays, we found that VEGFR2 binds to the Sp1-responsive region of the VEGFR2 proximal promoter. These results were confirmed by EMSA assays, using the same region of the VEGFR2 promoter. Importantly, we show that the VEGFR2 DNA binding is directly linked to the transcriptional activation of the VEGFR2 promoter. By reporter assays, we found that the region between -300/-116 relative to the transcription start site is essential to confer VEGFR2-dependent transcriptional activity. It was previously described that nuclear translocation of the VEGFR2 is dependent on its activation by VEGF. In agreement, we observed that the binding of VEGFR2 to DNA requires VEGF activation, being blocked by Bevacizumab and Sunitinib, two anti-angiogenic agents that inhibit VEGFR2 activation. Our findings demonstrate a new mechanism by which VEGFR2 activates its own promoter that could be involved in amplifying the angiogenic response
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