Robust industrial Saccharomyces cerevisiae strains for very high gravity bio-ethanol fermentations

The application and physiological background of two industrial Saccharomyces cerevisiae strains, isolated from harsh industrial environments, were studied in Very High Gravity (VHG) bio-ethanol fermentations. VHG laboratory fermentations, mimicking industrially relevant conditions, were performed with PE-2 and CA1185 industrial strains and the CEN.PK113-7D laboratory strain. The industrial isolates produced remarkable high ethanol titres (>19%, v/v) and accumulated an increased content of sterols (2 to 5-fold), glycogen (2 to 4-fold) and trehalose (1.1-fold), relatively to laboratory strain. For laboratory and industrial strains, a sharp decrease in the viability and trehalose concentration was observed above 90 g l-1 and 140 g l-1 ethanol, respectively. PE-2 and CA1185 industrial strains presented important physiological differences relatively to CEN.PK113-7D strain and showed to be more prepared to cope with VHG stresses. The identification of a critical ethanol concentration above which viability and trehalose concentration decrease significantly is of great importance to guide VHG process engineering strategies. This study contributes to the improvement of VHG processes by identifying yeast isolates and gathering yeast physiological information during the intensified fermentation process, which, besides elucidating important differences between these industrial and laboratory strains, can drive further process optimization.The authors thank Daniel Gomes for performing some of the fermentation samples analyses, COPAM - Companhia Portuguesa de Amidos S.A. (Portugal) for kindly providing the CSL, and Rosane Schwan (Federal University of Lavras, Brazil) for kindly providing the yeast strains PE-2 and CA1185. The financial support of Fundacao para a Ciencia e a Tecnologia (FCT), Portugal, is acknowledged: project ProBioethanol PTDC/BIO/66151/2006, grant SFRH/BD/64776/2009 to F.B. Pereira and grant SFRH/BPD/44328/2008 to P.M.R. Guimaraes

Optimization of low-cost medium for very high gravity ethanol fermentations by Saccharomyces cerevisiae using statistical experimental designs

Statistical experimental designs were used to develop a medium based on corn steep liquor (CSL) and other low-cost nutrient sources for high-performance very high gravity (VHG) ethanol fermentations by Saccharomyces cerevisiae. The critical nutrients were initially selected according to a Plackett–Burman design and the optimized medium composition (44.3 g/L CSL; 2.3 g/L urea; 3.8 g/L MgSO4·7H2O; 0.03 g/L CuSO4·5H2O) for maximum ethanol production by the laboratory strain CEN.PK 113-7D was obtained by response surface methodology, based on a three-level four-factor Box-Behnken design. The optimization process resulted in significantly enhanced final ethanol titre, productivity and yeast viability in batch VHG fermentations (up to 330 g/L glucose) with CEN.PK113-7D and with industrial strain PE-2, which is used for bio-ethanol production in Brazil. Strain PE-2 was able to produce 18.6 ± 0.5% (v/v) ethanol with a corresponding productivity of 2.4 ± 0.1 g/L/h. This study provides valuable insights into cost-effective nutritional supplementation of industrial fuel ethanol VHG fermentations.The authors thank Marisa Cunha for performing the preliminary fermentations with varying CSL concentrations, COPAM - Companhia Portuguesa de Amidos, S.A. (Portugal) for kindly providing glucose syrup and CSL, and Rosane Schwan (University of Lavras, Brazil) for kindly providing the yeast strain PE-2. The financial support of Fundacao para a Ciencia e a Tecnologia (FCT), Portugal, is acknowledged: Project ProBioethanol PTDC/BIO/66151/2006, Grant SFRH/BD/64776/2009 to F.B. Pereira and Grant SFRH/BPD/44328/2008 to P.M.R. Guimaraes

Imidazole processing of wheat straw and eucalyptus residues: comparison of pre-treatment conditions and their influence on enzymatic hydrolysis

ABSTRACT: Biomass pre-treatment is a key step in achieving the economic competitiveness of biomass conversion. In the present work, an imidazole pre-treatment process was performed and evaluated using wheat straw and eucalyptus residues as model feedstocks for agriculture and forest-origin biomasses, respectively. Results showed that imidazole is an efficient pre-treatment agent; however, better results were obtained for wheat straw due to the recalcitrant behavior of eucalyptus residues. The temperature had a stronger effect than time on wheat straw pre-treatment but at 160 degrees C and 4 h, similar results were obtained for cellulose and hemicellulose content from both biomasses (ca. 54% and 24%, respectively). Lignin content in the pre-treated solid was higher for eucalyptus residues (16% vs. 4%), as expected. Enzymatic hydrolysis, applied to both biomasses after different pre-treatments, revealed that results improved with increasing temperature/time for wheat straw. However, these conditions had no influence on the results for eucalyptus residues, with very low glucan to glucose enzymatic hydrolysis yield (93% for wheat straw vs. 40% for eucalyptus residues). Imidazole can therefore be considered as a suitable solvent for herbaceous biomass pre-treatment.info:eu-repo/semantics/publishedVersio

Cell recycling during repeated very high gravity bio-ethanol fermentations using the industrial Saccharomyces cerevisiae strain PE-2

A very high gravity (VHG) repeatedbatch fermentation system using an industrial strain of Saccharomyces cerevisiae PE-2 (isolated from sugarcane-to-ethanol distillery in Brazil) and mimicking industrially relevant conditions (high inoculation rates and low O2 availability) was successfully operated during fifteen consecutive fermentation cycles, attaining ethanol at 17.1 ± 0.2% (v/v) with a batch productivity of 3.5 ± 0.04 g l-1 h-1. Moreover, this innovative operational strategy (biomass refreshing step) prevented critical decreases on yeast viability levels and promoted high accumulation of intracellular glycerol and trehalose, which can provide an adaptive advantage to yeast cells under harsh industrial environments. This study contributes to the improvement of VHG fermentation processes by exploring an innovative operational strategy that allows attaining very high ethanol titres without a critical decrease of the viability level thus minimizing the production costs due to energy savings during the distillation process.The authors thank COPAM-Companhia Portuguesa de Amidos SA (Portugal) for kindly providing the CSL, and Rosane Schwan (Federal University of Lavras, Brazil) for kindly providing the PE-2 yeast strain. The financial support of Fundacao para a Ciencia e a Tecnologia (FCT), Portugal, is acknowledged: project ProBioethanol PTDC/BIO/66151/2006, grant SFRH/BD/64776/2009 to F. B. Pereira and grant SFRH/BPD/44328/2008 to P. M. R. Guimaraes

Plasmid-mediate transfer of FLO-1 into industrial Saccharomyces cerevisiae PE-2 strain creates a strain useful for repeat-batch fermentations involving flocculation-sedimentation

The flocculation gene FLO1 was transferred into the robust industrial strain Saccharomyces cerevisiae PE-2 by the lithium acetate method. The recombinant strain showed a fermentation performance similar to that of the parental strain. In 10 repeat-batch cultivations in VHG medium with 345 g glucose/L and cell recycling by flocculation–sedimentation, an average final ethanol concentration of 142 g/L and an ethanol productivity of 2.86 g/L/h were achieved. Due to the flocculent nature of the recombinant strain it is possible to reduce the ethanol production cost because of lower centrifugation and distillation costs.The authors thank COPAM - Companhia Portuguesa de Amidos S.A. (Portugal) for kindly providing the CSL, Rosane Schwan (Federal University of Lavras, Brazil) for kindly providing the PE-2 yeast strain and Merja Penttila (VTT, Finland) for plasmids pET13.1 and pBR-ADH1-FL01. The financial support of Fundacao para a Ciencia e a Tecnologia (FCT), Portugal, is acknowledged. Project ProBioethano

Multiomics implicate gut microbiota in altered lipid and energy metabolism in Parkinson's disease

We aimed to investigate the link between serum metabolites, gut bacterial community composition, and clinical variables in Parkinson's disease (PD) and healthy control subjects (HC). A total of 124 subjects were part of the study (63 PD patients and 61 HC subjects). 139 metabolite features were found to be predictive between the PD and Control groups. No associations were found between metabolite features and within-PD clinical variables. The results suggest alterations in serum metabolite profiles in PD, and the results of correlation analysis between metabolite features and microbiota suggest that several bacterial taxa are associated with altered lipid and energy metabolism in PD.Peer reviewe

Serotonin and tryptophan metabolites, autoantibodies and gut microbiome in APECED

Objective: Intestinal autoimmunity with gastrointestinal (GI) dysfunction has been shown in patients with autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED). Patients lack entero-endocrine (EE) cells and have circulating autoantibodies (Aabs) against critical enzymes in serotonin (5-HT) biosynthesis. Design: We sought to determine the serum levels of 5-HT, tryptophan (Trp) metabolites and L-DOPA in 37 Finnish APECED patients and to correlate their abundance with the presence of TPH and AADC Aabs, GI dysfunction and depressive symptoms. We also performed an exploratory analysis of the gut microbiome. Methods: Serum 5-HT, L-DOPA and Trp metabolite levels were determined by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). TPH and AADC Aabs were measured by ELISA. Depression was assessed with a structured RBDI questionnaire. The V3-V4 regions of the bacterial 16S rRNA gene were sequenced for gut microbiome exploration. Results: Serum 5-HT levels were significantly decreased (130 +/- 131 nmol/L vs 686 +/- 233 nmol/L, P <0.0001) in APECED patients with TPH-1 (+/- AADC) Aabs compared to controls and patients with only AADC Aabs. Reduced 5-HT levels correlated with constipation. The genus Escherichia/Shigella was overrepresented in the intestinal microbiome. No correlation between serum Trp, 5-HT or L-DOPA levels and the RBDI total score, fatigue or sleep disorders was found. Conclusions: This exploratory study found low serum levels of 5-HT to be associated with constipation and the presence of TPH-1 and AADC Aabs, but not with symptoms of depression. Hence, serum 5-HT, TPH1 and AADC Aabs should be determined in APECED patients presenting with GI symptoms.Peer reviewe

Gut microbiota in Parkinson's disease : Temporal stability and relations to disease progression

Background: Several publications have described differences in cross-sectional comparisons of gut microbiota between patients with Parkinson's disease and control subjects, with considerable variability of the reported differentially abundant taxa. The temporal stability of such microbiota alterations and their relationship to disease progression have not been previously studied with a high-throughput sequencing based approach. Methods: We collected clinical data and stool samples from 64 Parkinson's patients and 64 control subjects twice, on average 2.25 years apart. Disease progression was evaluated based on changes in Unified Parkinson's Disease Rating Scale and Levodopa Equivalent Dose, and microbiota were characterized with 16S rRNA gene amplicon sequencing. Findings: We compared patients to controls, and patients with stable disease to those with faster progression. There were significant differences between microbial communities of patients and controls when corrected for confounders, but not between timepoints. Specific bacterial taxa that differed between patients and controls at both timepoints included several previously reported ones, such as Roseburia, Prevotella and Bifidobacterium. In progression comparisons, differentially abundant taxa were inconsistent across methods and timepoints, but there was some support for a different distribution of enterotypes and a decreased abundance of Prevotella in faster-progressing patients. Interpretation: The previously detected gut microbiota differences between Parkinson's patients and controls persisted after 2 years. While we found some evidence for a connection between microbiota and disease progression, a longer follow-up period is required to confirm these findings. (C) 2019 The Authors. Published by Elsevier B.V.Peer reviewe

On the nature of the (de)coupling of the magnetostructural transition in Er5_5Si4_4

In this report, a successful thermodynamical model was employed to understand the structural transition in Er$_5$Si$_4$, able to explain the decoupling of the magnetic and structural transition. This was achieved by the DFT calculations which were used to determine the energy differences at 0 K, using a LSDA+U approximation. It was found that the M structure as the stable phase at low temperatures as verified experimentally with a $\Delta F_0 = -$0.262 eV. Finally, it was achieved a variation of Seebeck coefficient ($\sim$ 6 $\mu$V) at the structural transition which allow to conclude that the electronic entropy variation is negligible in the transition.Comment: 17 pages, 3 figures, 1 tabl