Saccharomyces cerevisiae CEN.PK113-7D
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PROVIDER: PRJNA414371 | ENA |
REPOSITORIES: ENA
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Project description:L-Arabinose occurs at economically relevant levels in lignocellulosic hydrolysates. Especially at low concentrations of L-arabinose, its uptake via the Gal2 galactose transporter is an important rate-controlling step in the complete conversion of these feedstocks by engineered, pentose-metabolizing Saccharomyces cerevisiae strains. Chemostat-based transcriptome analysis yielded 16 putative sugar transporter genes in the filamentous fungus Penicillium chrysogenum whose transcript levels were at least three-fold higher in L-arabinose-limited cultures than in glucose-limited and ethanol-limited cultures. Of five genes that showed an over 30-fold higher transcript level in arabinose-grown cultures, only one (Pc20g01790) restored growth on L-arabinose upon expression in an engineered L-arabinose-fermenting S. cerevisiae strain in which GAL2 had been deleted. Sugar-transport assays indicated that Pc20g01790this transporter, designated as PcAraT, encodes functions as a high-affinity (Km = 0.13 mM) L-arabinose-proton symporter that does not transport xylose or glucose. An L-arabinose-metabolizing S. cerevisiae strain co-expressing Pc20g01790PcAraT and GAL2 showed lower residual substrate concentrations in L-arabinose-limited chemostat cultures (4 mg L-1) than a congenic strain in which L-arabinose import exclusively depended on Gal2 (1.8 g L-1). Inhibition of L-arabinose transport by these sugars was less pronounced than observed with Gal2. A hexose-phosphorylation-deficient, L-arabinose-metabolizing S. cerevisiae strain expressing PcAraT Pc20g0190 grew on 20 g L-1 L-arabinose in the presence of 20 g L-1 glucose, which completely inhibited growth on L-arabinose of a congenic strain dependent on L-arabinose transport via Gal2. Its high affinity and specificity for L-L-arabinose, combined with limited sensitivity to inhibition by glucose and D-D-xylose make PcAraT/ Pc20g01790 a valuable transporter gene for application in metabolic engineering strategies aimed at engineering S. cerevisiae strains for efficient conversion of lignocellulosic hydrolysates.
2017-10-03 | GSE104491 | GEO
Project description:In presence of high concentrations of L-arabinose, Vibrio cholerae enters into a non-proliferative state. In V. cholerae, when L-arabinose in present into the media, it is incorporated and metabolized through the galactose pathway. In the present datasets we show that in presence of L-arabinose, transposon insertions in the galactose pathway confer a resistance to the detrimental effect of L-arabinose.
2020-11-16 | E-MTAB-9747 | biostudies-arrayexpress
Project description:Investigation of whole genome gene expression levels in Clostridium acetobutylicum strain 824 grown individually on starch, galactose, lactose, mannose, fructose, glucose, arabinose, xylose, cellobiose, maltose, sucrose
2010-05-15 | GSE18471 | GEO
Project description:Arabinose and galactose are major components of both particulate and dissolved organic matter. They often constitute polysaccharides such as arabinogalactan. Field studies have demonstrated a turnover of arabinogalactan, but enzymes and organisms involved have not been explored. In this study, the degradation of arabinogalactan by the marine flavobacterium Maribacter sp. MAR_2009_72 was investigated with growth experiments and proteomic analyses. Growth of Maribacter sp. MAR_2009_72 on arabinogalactan displayed its ability to utilize arabinogalactan. Proteomic analysis of cells grown on arabinogalactan, arabinose, galactose or glucose revealed expression of specific proteins in presence of arabinogalactan, mainly glycoside hydrolases. Candidates for extracellular glycan hydrolysis are five alpha-L-arabinofuranosidases affiliating with glycosyl hydrolase (GH) families 43 and 51, four unsaturated rhamnogalacturonyl hydrolases (GH105) and a protein with a glycosyl hydrolase family like domain. We detected expression of three induced TonB-dependent SusCD transporter systems, one SusC and nine glycosyl hydrolases with a predicted periplasmatic location. These are affiliated with the families GH3, GH10, GH29, GH31, GH67, GH78 and GH115. The genes are located within three canonical polysaccharide utilization loci (PUL), but also outside of the defined loci. PULs can be classified as specific for arabinogalactan, galactose-containing glycans and for arabinose-containing glycans. The breadth of enzymatic functions expressed in Maribacter sp. MAR_2009_72 as response to arabinogalactan from terrestrial plant larch suggests that Flavobacteriia are main catalysts of the rapid turnover of arabinogalactans in the marine environment.
2024-05-02 | PXD049074 | Pride
Project description:Alkaline hemicellulytic bacteria Bacillus sp. N16-5 has abroad substrate spectrum and exhibits great growth ability on complex carbohydrates. In order to get insight into its carbohydrate utilization mechanism, global transcriptional profiles were separately determined for growth on glucose, fructose, mannose, galactose, arabinose, xylose, galactomannan, xylan, pectin and carboxymethyl cellulose by using one-color microarrays. Substrate induced gene expression was measured when culture was grown on glucose, fructose, mannose, galactose, arabinose, xylose, galactomannan, xylan and CMC to mid-logarithmic phase.
2014-06-17 | E-GEOD-38276 | biostudies-arrayexpress
Project description:Background: Bacterial fermentation of carbohydrates from sustainable lignocellulosic biomass into biofuels by the anaerobic bacterium Clostridium acetobutylicum is a promising alternative energy source to fossil fuels. Understanding the complex metabolic pathways it employs and how they are regulated will contribute to improved biofuel production. Recently, it has been demonstrated that xylose is not appreciably fermented in the presence of arabinose, suggesting a hierarchy of pentose utilization in this organism. Our goal is to uncover if transcriptional regulation contributes to this hierarchy. Results: Growth and sugar consumption rates showed that arabinose, like glucose, actively represses xylose utilization in cultures fermenting xylose. RNA-Seq revealed there was a large overlap in differentially regulated genes after addition of arabinose or glucose, suggesting a common mechanism of regulation. A putative ORF was identified that may be important for transcriptional regulation in response to the nutritional state of the cells. Conclusions: Decreased xylose consumption, increased acetate production, and transcription of the phosphoketolase gene (CA_C1343) revealed a transition of pentose catabolism from the pentose phosphate pathway to the phosphoketolase pathway after addition of arabinose. Together, these results substantiate the claim that arabinose is utilized preferentially over xylose in C. acetobutylicum. Furthermore, they demonstrate that this phenomenon is modulated in part at the transcriptional level, and they provide valuable insight into potential mechanisms for altering pentose utilization to modulate fermentation products for biofuel production.
2017-12-20 | GSE107804 | GEO
Project description:Alkaline hemicellulytic bacteria Bacillus sp. N16-5 has abroad substrate spectrum and exhibits great growth ability on complex carbohydrates. In order to get insight into its carbohydrate utilization mechanism, global transcriptional profiles were separately determined for growth on glucose, fructose, mannose, galactose, arabinose, xylose, galactomannan, xylan, pectin and carboxymethyl cellulose by using one-color microarrays.
2014-06-17 | GSE38276 | GEO
Project description:Investigation of whole genome gene expression levels in Clostridium acetobutylicum strain 824 grown individually on starch, galactose, lactose, mannose, fructose, glucose, arabinose, xylose, cellobiose, maltose, sucrose A 33 array study using total RNA recovered from three separate wild-type cultures of Clostridium acetobutylicum 824 grown on each of the following sugars: starch, galactose, lactose, mannose, fructose, glucose, arabinose, xylose, cellobiose, maltose, sucrose. RNA extraction, cDNA production, and labeling were performed independently on three separate cultures for each carbohydrate. Each independent sample was hybridized to one array equating to one technical replicate for each biological sample and three biological replicates for each condition tested. Each array contains 2 replicates of 9 probes approximately 60 bp in length for each of the 3,842 open reading frames annotated in the C. acetobutylicum 824 genome.
2010-05-15 | E-GEOD-18471 | biostudies-arrayexpress
Project description:The haploid Saccharomyces cerevisiae strain CEN.PK113-7D is a popular model system for metabolic engineering and systems biology research. Current genome assemblies are based on short-read sequencing data scaffolded based on homology to strain S288C. However, these assemblies contain large sequence gaps, particularly in subtelomeric regions, and the assumption of perfect homology to S288C for scaffolding introduces bias. In this study, we obtained a near-complete genome assembly of CEN.PK113-7D using only Oxford Nanopore Technology's MinION sequencing platform. Fifteen of the 16 chromosomes, the mitochondrial genome and the 2-μm plasmid are assembled in single contigs and all but one chromosome starts or ends in a telomere repeat. This improved genome assembly contains 770 Kbp of added sequence containing 248 gene annotations in comparison to the previous assembly of CEN.PK113-7D. Many of these genes encode functions determining fitness in specific growth conditions and are therefore highly relevant for various industrial applications. Furthermore, we discovered a translocation between chromosomes III and VIII that caused misidentification of a MAL locus in the previous CEN.PK113-7D assembly. This study demonstrates the power of long-read sequencing by providing a high-quality reference assembly and annotation of CEN.PK113-7D and places a caveat on assumed genome stability of microorganisms.
| S-EPMC5812507 | biostudies-literature
Project description:Trichoderma reesei is used to produce saccharifying enzyme cocktails for biofuels. There is limited understanding of the transcription factors (TFs) that regulate the release and catabolism of L-arabinose and D-galactose, as the main TF XYR1 is only partially involved. The T. reesei ortholog of ARA1 from Pyricularia oryzae that regulates L-arabinose release and catabolism, was deleted and characterized by growth profiling and transcriptomics along with a xyr1 mutant and xyr1/ara1 double mutant. Our results show that, in addition to the L-arabinose-related role, T. reesei ARA1 is essential for D-galactose release and catabolism, while XYR1 is not involved in this process.
2017-11-02 | GSE104606 | GEO