Saccharomyces cerevisiae stress responses to HMF and furfural (xylose and glucose) Oct
ABSTRACT: HMF and furfural were pulse added to xylose-utilizing Saccharomyces cerevisiae during either the glucose consumption phase or the xylose consumption phase. Transcriptome samples were collected before and one hour after pulsing of inhibitors. Three biological replicates from each conditions analyzed.
Project description:The inhibitors hydroxymethylfurfural (HMF) and furfural were added to the feed-medium of carbon-limited anaerobic chemostat cultures. Samples were taken for transcriptome analysis at steady-state from cultures with inhibitors and without inhibitors. Three biological replicates from each condition (inhibitors, no inhibitors) were analyzed.
Project description:Adapted tolerant yeast strain Y-50049 is able to in situ detoxify furfural and HMF while the wild type control Y-12632 repressed to loss function under challenges of 20 mM each of furfural and HMF A time course study during the lag phase with cells harvested at 18, 24, 28, and 42 h after 20 mM furfural and 20 mM HMF treatment
Project description:The data explore the transcriptional response of strain LY180 and the furfural-resistant derivative EMFR9 to 0.5 g/L furfural LY180 and EMFR9 and differences in their expression profiles are described in Miller, E. N., L. R. Jarboe, L. P. Yomano, S. W. York, K. T. Shanmugam, and L. O. Ingram. 2009. Silencing of NADPH-dependent oxidoreductase genes (yqhD and dkgA) in furfural-resistant ethanologenic Escherichia coli. Appl Environ Microbiol 75:4315-23. The response of LY180 to furfural is described in Miller, E. N., L. R. Jarboe, P. C. Turner, P. Pharkya, L. P. Yomano, S. W. York, D. Nunn, K. T. Shanmugam, and L. O. Ingram. 2009. Furfural Inhibits Growth by Limiting Sulfur Assimilation in Ethanologenic Escherichia coli strain LY180. Appl Environ Microbiol., 2009. Total RNA was prepared from cultures of LY180 and EMFR9 immediately before and 15 min after addition of furfural to 0.5 g/L. The Nimblegen TI83333 chip measures expression of 4,237 genes, with 5 replicates, and 18 probes average per gene. Data from replicate 1 (OID 8641) and replicate 2 (OID 14117) was analyzed independently.
Project description:Cellular tolerance toward furfural is a complex phenotype involved many genes, and hard to be improved by manipulating individual genes. We previously established exogenous global regulator IrrE mutants that confer Escherichia coli with significantly enhanced tolerance to furfural stress. In order to elucidate the mechanism for enhancement of furfural tolerance in the mutants and to identify new genes and pathways that can be possible targets for engineering of furfural tolerance, we carried out comparative transcriptomic with the representative strains F1-37 and WT (harboring the furfural-tolerant mutant F1-37 of IrrE and the wild type IrrE, respectively). The data from transcriptome analyses were deposited here. Cells of furfural-tolerant mutant F1-37 and wild-type strain WT were grown in LB medium supplemented with furfural, and the cells were harvested in the exponential phase. The samples for both of these two strains were prepared in triplicate with biological replicates.
Project description:Furfural is the prevalent microbial inhibitor generated during pretreatment and hydrolysis of lignocellulosic biomass to monomeric sugars, but the molecular response of Clostridium beijerinckii NCIMB 8052 to this compound is unknown. To discern the effect of furfural on C. beijerinckii and to gain insights into the molecular mechanisms of action and detoxification, we studied the physiological changes of furfural-stressed cultures during acetone-butanol-ethanol (ABE) fermentation, and profiled differentially expressed genes by genome-wide transcriptional analysis. C. beijerinckii exposed to furfural stress during the acidogenic growth phase produced 13% more ABE than the unstressed control. The growth and ABE by C. beijerinckii ceased following exposure to furfural stress during the solventogenic growth phase. By comparing gene expression of furfural-stressed cultures to that of the unstressed control, at both the acidogenic and solventogenic phases, we ascertained that furfural induces expression of several genes including those that code for heat shock proteins, redox enzymes and cofactor associated proteins, and ATP-binding cassette transporters, and represses genes belonging to the phosphotransferase system, two-component system, chemotaxis and cell motility. Based on these results, we discuss the underpinning for furfural-mediated change in ABE fermentation by the solventogenic Clostridium species. C. beijerinckii 8052 pre-culture was incubated anaerobically to attain acidogenic or solventogenic growth phase. The culture was then subdivided into two bottles. One bottle was challenged with furfural and the other bottle was left unchallenged. After 3 h growth, C. beijerinckii 8052 samples were collected, during which the original concentration of furfural in the growth medium was reduced to more than half. Total RNA was isolated, purified, converted to enriched mRNA, and dye-coupled (Alexa Fluor 555) complementary cRNA. This was followed by hybridization and microarray data analysis.
Project description:To understrand the altered global gene expression levels in C. glutamicum wild type in presence of furfural, transcriptome profiling was performed. Transcriptome profiles of the wild type grown in CgXII medium without furfural and with furfural stresses (each 6.5 mM, 13 mM, and 20 mM) were compared by using the samples taken at the OD600 of 6 (for the control and experiments). Each experiment was performed with a duplicate.
Project description:The data explore the transcription of strain LY180 and the yqhC deletion mutant LY180 del yqhC without and with 0.5 g/L furfural. LY180 and LY180 del yqhC are described in Turner, PC, EN Miller, LR Jarboe, P Pharkya, KT Shanmugam, and LO Ingram. 2009. Escherichia coli YqhC regulates transcription of the adjacent yqhD and dkgA genes, and mutations in yqhC contribute to furfural resistance in ethanologenic strains (in preparation for submission to Appl Env Microbiol) Total RNA was prepared from cultures of LY180 and LY180 del yqhC immediately before and 15 min after addition of furfural to 0.5 g/L. The Nimblegen TI83333 chip measures expression of 4,237 genes, with 5 replicates, and 18 probes average per gene.
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. Overall design: 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 (0min before furfural shock, 15min and 60min post-furfural shock) 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. Three biological replicates were used for each condition.
Project description:Investigation of the expression profiling of the ethanologenic Zymomonas mobilis in response to furfural stress. A six chip study using total RNA recovered from three separate wild-type cultures of Zymomonas mobilis ATCC31821 and three separate cultures of a triple treated with 1.0 g/l furfural. Each chip measures the expression level of 1800 genes from Zymomonas mobilis ATCC31821 and the associated plasmids, with three-fold technical redundancy.
Project description:This SuperSeries is composed of the following subset Series: GSE28374: DNA methylation of miRNA genes in HMEC and HMF GSE28375: Histone H3 acetylation of miRNA genes in HMEC and HMF GSE28376: Histone H3 lysine 27 trimethylation of miRNA genes in HMEC and HMF GSE28377: Histone H3 lysine 4 trimethylation of miRNA genes in HMEC and HMF GSE28378: Histone H3 lysine 9 dimethylation of miRNA genes in HMEC and HMF Refer to individual Series