Genomic and phenotypic characterization of a refactored xylose utilizing Saccharomyces cerevisiae for lignocellulosic biofuels/biochemicals production
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ABSTRACT:
PROVIDER: PRJNA476957 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR7405150_1.fastq.gz | Fastqsanger.gz | |||
| SRR7405150_2.fastq.gz | Fastqsanger.gz | |||
| SRR7405151_1.fastq.gz | Fastqsanger.gz | |||
| SRR7405151_2.fastq.gz | Fastqsanger.gz | |||
| SRR7405146_1.fastq.gz | Fastqsanger.gz |
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Similar Datasets
Project description:To understand the mechanisms of enhanced xylose metabolism of XUSE, we reported transcriptome analysis of XUS (rationally engineered strain) and XUSE (evolved XUS strain) grown on xylose by performing RNA-sequencing. Compared with XUS, XUSE showed significantly different gene expression landscape with 463 up-regulated and 675 down-regulated genes (>2-fold, p-value<0.05). While the genes involved in non-oxidative pentose phosphate pathway, such as TKL1, TKL2 and TAL1, were down-regulated in XUSE, it shows the decreased transcriptional patterns of a number of genes encoding TCA cycle and respiratory enzymes. Also, the genes involved in the cellular metal ion regulation and sugar transporters showed significantly changed expression levels in XUSE. This study shows the gene expression landscape of XUSE revealing engineering strategies for the enhanced xylose metabolism in S. cerevisiae.
2018-12-26 | GSE116076 | GEO
Project description:Background:Engineered strains of Saccharomyces cerevisiae have significantly improved the prospects of biorefinery by improving the bioconversion yields in lignocellulosic bioethanol production and expanding the product profiles to include advanced biofuels and chemicals. However, the lignocellulosic biorefinery concept has not been fully applied using engineered strains in which either xylose utilization or advanced biofuel/chemical production pathways have been upgraded separately. Specifically, high-performance xylose-fermenting strains have rarely been employed as advanced biofuel and chemical production platforms and require further engineering to expand their product profiles. Results:In this study, we refactored a high-performance xylose-fermenting S. cerevisiae that could potentially serve as a platform strain for advanced biofuels and biochemical production. Through combinatorial CRISPR-Cas9-mediated rational and evolutionary engineering, we obtained a newly refactored isomerase-based xylose-fermenting strain, XUSE, that demonstrated efficient conversion of xylose into ethanol with a high yield of 0.43 g/g. In addition, XUSE exhibited the simultaneous fermentation of glucose and xylose with negligible glucose inhibition, indicating the potential of this isomerase-based xylose-utilizing strain for lignocellulosic biorefinery. The genomic and transcriptomic analysis of XUSE revealed beneficial mutations and changes in gene expression that are responsible for the enhanced xylose fermentation performance of XUSE. Conclusions:In this study, we developed a high-performance xylose-fermenting S. cerevisiae strain, XUSE, with high ethanol yield and negligible glucose inhibition. Understanding the genomic and transcriptomic characteristics of XUSE revealed isomerase-based engineering strategies for improved xylose fermentation in S. cerevisiae. With high xylose fermentation performance and room for further engineering, XUSE could serve as a promising platform strain for lignocellulosic biorefinery.
| S-EPMC6162923 | biostudies-literature
Project description:Xylose-utilizing yeasts with tolerances to fermentation inhibitors (such as weak organic acids) and high temperature are needed for cost-effective simultaneous saccharification and co-fermentation (SSCF) of lignocellulosic materials. We constructed a novel xylose-assimilating Saccharomyces cerevisiae strain with improved fermentation performance under heat and acid co-stress using the genome shuffling technique. Two xylose-utilizing diploid yeasts with different genetic backgrounds were used as the parental strains for genome shuffling. The hybrid strain Hyb-8 showed significantly higher xylose fermentation ability than both parental strains (Sun049T-Z and Sun224T-K) under co-stress conditions of heat and acids. To screen for genes that might be important for fermentation under heat and acid co-stress, a transcriptomic analysis of hybrid strain Hyb-8 and its parental strains was performed.
2017-07-25 | GSE101786 | GEO
Project description:Xylose induced effects on metabolism and gene expression during anaerobic growth of an engineered Saccharomyces cerevisiae on mixed glucose-xylose medium were quantified. Gene expression of S. cerevisiae harbouring an XR-XDH pathway for xylose utilisation was analysed from early cultivation when mainly glucose was metabolised, to times when xylose was co-consumed in the presence of low glucose concentrations, and finally, to glucose depletion and solely xylose being consumed. Cultivations on glucose as a sole carbon source were used as a control. Genome-scale dynamic flux balance analysis models were developed and simulated to analyse the metabolic dynamics of S. cerevisiae in the cultivations. Model simulations quantitatively estimated xylose dependent dynamics of fluxes and challenges to the metabolic network utilisation. Increased relative xylose utilisation was predicted to induce two-directionality of glycolytic flux and a redox challenge already at low glucose concentrations. Xylose effects on gene expression were observed also when glucose was still abundant. Remarkably, xylose was observed to specifically delay the glucose-dependent repression of particular genes in mixed glucose-xylose cultures compared to glucose cultures. The delay occurred during similar metabolic flux activities in the both cultures. Xylose is abundantly present together with glucose in lignocellulosic streams that would be available for the valorisation to biochemicals or biofuels. Yeast S. cerevisiae has superior characteristics for a host of the bioconversion except that it strongly prefers glucose and the co-consumption of xylose is yet a challenge. Further, since xylose is not a natural substrate of S. cerevisiae, the regulatory response it induces in an engineered yeast strain cannot be expected to have evolved for its utilisation. Dynamic cultivation experiments on mixed glucose-xylose medium having glucose cultures as control integrated with mathematical modelling allowed to resolve specific effects of xylose on the gene expression and metabolism of engineered S. cerevisiae in the presence of varying amounts of glucose.
2016-05-24 | GSE69966 | GEO
Project description:We have identified a novel xylose-utilization phenotype controlled by a single gene, and we sought to characterize how both the presence and absence of xylose in the presence or absence of the trait affected the S. cerevisiae transcriptional program over time. To do so, we measured global steady state mRNA levels on three pairs of sister spores from a Simi White strain that was backcrossed twice to S288c. Each pair of spores was from an independent tetrad, and contained one xylose utilizing ("positive", GSY2465, 2466, 2469) and one xylose non-utilizing spore ("negative", GSY2464, 2467, 2470). We grew each of the six spores in the presence or absence of xylose for 72 hours with samples taken every 8 hours. We then assayed relative RNA abundance versus a pooled reference, containing equimolar amounts of each sample, using Agilent yeast catalog arrays.
2010-04-09 | GSE19116 | GEO
Project description:Background: Lignocellulosic biomass is a promising renewable feedstock for the microbial production of fuels. To release the major fermentable sugars such as glucose and xylose, pretreatment, hydrolysis, and subsequent conditioning of biomass feedstock are needed. During this process, many toxic compounds are produced or introduced which subsequently inhibit microbial growth and in many cases the production titer and rate. An understanding of the toxic effects of compounds found in hydrolysate on the fermentation microorganism is critical to improving biofuel yields in the process. One of the inhibitory compounds is furfural, liberated from hemicelluloses, which strongly inhibits the cell growth and ethanol production especially from xylose. Zymomonas mobilis is a capable ethanologenic bacterium with high ethanol productivity and high levels of ethanol tolerance. The development of robust biocatalyst to tolerate the lignocellulosic pretreatment inhibitors is one of the key elements for economic biofuel production. Results: In this study, the molecular responses of Z. mobilis to furfural, one major pretreatment inhibitor, were investigated using transcriptomic approaches of chip-based microarray. Furfural shock time course experiment with 3 g/L furfural supplemented when cells reach exponential phase and stress response experiment in the presence of 2 g/L furfural from the beginning of fermentation were carried out to study the short and long-term effect of furfural on 8b physiological and transcriptional profiles. The presence and supplementation of furfural negatively affect 8b growth in terms of final biomass and the fermentation time. Transcriptomic studies indicated that the response of 8b to furfural is dynamic, complex and differences exist between short-term shock response and long-term stress response. However, the gene function categories are similar with most downregulated genes related to translation and biosynthesis, while the furfural-upregulated genes were mostly related to cellular processes of general stress response and energy metabolism. Conclusions: Similar to previous report that acetate inhibited the growth of Z. mobilis 8b in RM using glucose or xylose as carbon source, the existence or supplementation of another major hydrolysate inhibitor furfural also inhibited 8b growth with slowing the substrate utilization and ethanol production. The difference between carbon sources is more dramatic than that of the major hydrolysate inhibitors of both NH4OAc (GSE57553) and furfural (this study). Several gene targets have been selected for genetic studies with promising preliminary results.
2016-11-18 | GSE63540 | GEO
Project description:As advances are made toward the industrial feasibility of mass-producing biofuels and commodity chemicals with sugar-fermenting microbes, high feedstock costs continue to inhibit commercial application. Hydrolyzed lignocellulosic biomass represents an ideal feedstock for these purposes as it is cheap and prevalent. However, many microbes, including Escherichia coli, struggle to efficiently utilize this mixture of hexose and pentose sugars due to the regulation by the carbon catabolite repression (CCR) system. CCR causes a sequential utilization of sugars, rather than simultaneous utilization, resulting in reduced carbon yield and complex process implications in fed-batch fermentation. A mutation in the gene encoding the cyclic AMP receptor protein, Crp*, has been shown to disable CCR and improve co-utilization of mixed sugar substrates. Here, we present the strain construction and characterization of a site specific crp* chromosomal mutant in E. coli BL21 starTM (DE3). The crp* mutant strain demonstrates simultaneous consumption of glucose and xylose, suggesting a deregulated CCR system. The proteomic analysis further showed that cells link xylose consumption to energy production through the de novo nucleotide synthesis pathway, explaining the relatively slower growth of the crp* mutant strain. This highly characterized strain can be particularly beneficial for chemical production by simultaneously utilizing both C5 and C6 substrates from lignocellulosic biomass.
| MSV000092810 | MassIVE
Project description:We have identified a novel xylose-utilization phenotype controlled by a single gene, and we sought to characterize how both the presence and absence of xylose in the presence or absence of the trait affected the S. cerevisiae transcriptional program over time. To do so, we measured global steady state mRNA levels on three pairs of sister spores from a Simi White strain that was backcrossed twice to S288c. Each pair of spores was from an independent tetrad, and contained one xylose utilizing ("positive", GSY2465, 2466, 2469) and one xylose non-utilizing spore ("negative", GSY2464, 2467, 2470). We grew each of the six spores in the presence or absence of xylose for 72 hours with samples taken every 8 hours. We then assayed relative RNA abundance versus a pooled reference, containing equimolar amounts of each sample, using Agilent yeast catalog arrays. time_series_design
2010-04-09 | E-GEOD-19116 | biostudies-arrayexpress
Project description:The xylose fermentation rate during xylose consumption phase after glucose depleted in glucose-xylose co-fermentation (defined as GX stage) was much lower than that when xylose was the sole carbon source (defined as X stage). BSGX001 and XH7 are two engineered strains that have the xylose-utilizing capacity. Here,we investigate the transcriptional differences between GX stage and X stage of BSGX001 and XH7, respectively.
2018-04-25 | GSE95076 | GEO
Project description:The conversion of sugars in lignocellulosic hydrolysates to bioethanol represents an industrially relevant system for understanding microbial physiology associated with production of bio-based fuels and chemicals. To this end we have developed a new version of synthetic hydrolysate (SynH) modeled on highly concentrated 9% AFEX-pretreated cornstove hydrolysate (ACSH) with and without lignocellulose-derived inhibitors (LDIs) added, termed SynH3 and SynH3- respectively. We profiled the cellular responses of xylose-utilizing Z. mobilis 2032 grown in both SynH3- and SynH3 via collection and analysis of multiple omics-based data (multiomics) including transcriptomics, proteomics, and metabolomics. Our study was focused on answering the following two questions. First, how does Z. mobilis respond to LDIs in SynH3 and how does this compare to our previous studies with E. coli in 6% ACSH SynH? Second, what is the potential cause for the poor xylose conversion in the presence of LDIs? Addressing these questions will provide critical information for engineering of Z. mobilis strains with improved productivities in lignocellulosic hydrolysates.
2019-10-25 | GSE135718 | GEO