Lactate Stress in L. plantarum WCFS1
Ontology highlight
ABSTRACT:
ORGANISM(S): Lactobacillus plantarum
SUBMITTER: Douwe Molenaar
PROVIDER: E-GEOD-18354 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:Cultures were grown untill an optical density of 1.0. Cells were subsequently stressed with 2000 mM Na-lactate and samples were taken in time (15, 30, and 60 minutes).
2010-01-01 | GSE18354 | GEO
Project description:Transcriptome profiles of control Lactobacillus plantarum WCFS1 cells were compared with 8% ethanol adapted cells and with 10 min or 30 min 8% ethanol shocked cells. A continuous culture was performed in duplicate on medium containing ehtanol (tE). A shock experiment was performed in duplicate in which ethanol was added to a batch culture immediately after t0 (=control). The culture was sampled at time=0 (t0), time =10 min (t1) and time=30 min (t2).
2010-05-16 | E-GEOD-17847 | biostudies-arrayexpress
Project description:Transcriptome profiles of control Lactobacillus plantarum WCFS1 cells were compared with ctsR, hrcA, and ctsR-hrcA deletion mutants grown in MRS media at 28 and 40 degrees Celcius. Continuous cultures were performed in duplo on MRS media and whole genome transcriptome analysis was performed on the WCFS1 (WT), NZ3410CM (harboring a chlooramphenicol resistance marker (cat) replacement mutation in ctsR, dc), NZ3425CM (harboring a cat replacement mutation in hrcA, dh), and NZ3423 (harboring deletion mutation in ctsR and a cat replacement mutation inhrcA, dcdh) strains.
2012-01-01 | E-GEOD-31253 | biostudies-arrayexpress
Project description:Lactobacillus plantarum WCFS1 was grown under anaerobic carbon-limited conditions in a chemostat with complete biomass retention (retentostat). In this cultivation system, the biomass concentration progressively increases while the dilution rate is kept constant, resulting in decreased specific susbtrate availibility, and hence, a progressive decrease in the specific growth rate. During the progressive transition from growth to virtually no growth, the global changes occurring at the level of metabolism and gene expression were studied using a genome-scale metabolic model and DNA microarrays. Four different time-points are compared, corresponding to 4 different specific growth rates, and hence, 4 different ratios of energy used for maintenance and growth. The samples taken at the start of retentostat cultivation serves as a a reference sample, to which the three other samples (taken after 3 days, 17 days, and 31 days under retentostat conditions) are compared. No biological replicates: all samples were taken from the same retentostat fermentation.
2010-05-16 | E-GEOD-18340 | biostudies-arrayexpress
Project description:The level of the Central glycolytic gene regulator (CggR) was engineered in Lactobacillus plantarum NC8 and WCFS1 by over-expression and in-frame mutation of the cggR gene in order to evaluate its regulatory role on the glycolytic gap operon and the glyco The hybridization scheme of the different strains (wild-type, cggRKO mutant, cggR+) in relation to carbon source (glucose or ribose) has a loop design.
2010-05-16 | E-GEOD-18435 | biostudies-arrayexpress
Project description:Transcriptome profiles of control Lactobacillus plantarum WCFS1 cells were compared with a lp_1669 deletion mutant in MRS and 2*CDM media. A continuous culture was performed on MRS and 2*CDM media and whole genome transcriptome analysis was perfomred on the WCFS1 and NZ3417CM (harboring a chlooramphenicol resistance marker (cat) replacement mutation in lp_1669) strains.
2012-01-01 | E-GEOD-31254 | biostudies-arrayexpress
Project description:L. plantarum is known to possess an L-lactate inducible lactate racemase activity (Goffin et al. 2005. J. Bacteriol. 187:6750). In the present study, microarrays were used in order to identify all genes that are up-regulated by L-lactate, but not by a racemic mixture of D- and L-lactate. A mutant of L. plantarum NCIMB8826 deficient for NAD-dependent L-lactate activity (TF101; Ferain et al. 1994. 176:596), and thus producing no L-lactate, was grown in MRS medium at 28°C until mid-exponential phase (OD600nm 0.75). The culture was then divided into 3 sub-cultures. Optically pure sodium L-lactate (200 mM) was added to the first sub-culture (TF101 + L-lac 200 mM). An equimolar mixture of sodium D- and L-lactate (100 mM each) was added to the second sub-culture (TF101 + L/D-lac 200 mM). The third sub-culture was not treated (TF101; reference sample). The three sub-cultures were further incubated at 28°C for 1h30 (a time known to be sufficient for induction of lactate racemase activity by L-lactate). Cells were harvested by centrifugation. Microarray data were used ot identify genes that are specifically induced by L-lactate (comparison of TF101 with TF101 + L-lac 200 mM), but not by DL-lactate (comparison of TF101 with T101 + L/D-lac 200 mM). There are no biological replicates.
2013-07-01 | E-GEOD-43518 | biostudies-arrayexpress
Project description:Eubacterium limosum ATCC 8486 makes acetate and butyrate from various substrates and is found in the human intestine. The proteome of lactate-grown Eubacterium limosum was obtained in order to identify enzymes required for growth on this substrate, in particular to identify components that are unique to growth on lactate in comparison to other substrates for acetogenesis.
2019-07-25 | PXD013806 | Pride
Project description:This study aimed to characterise the transcriptomic response of the lactate-utilizing bacteria, Coprococcus catus and Anaerobutyricum soehngenii, grown on varying carbon sources. This work has allowed for identification of divergent gene clusters in each species contributing to the lactate utilisation pathway.
2021-04-18 | E-MTAB-10136 | biostudies-arrayexpress
Project description:Background: Evolutionary engineering is a powerful approach to isolate suppressor mutants and industrially relevant genotypes. Until recently, DNA microarray analysis was the only affordable genome-wide approach to identify the responsible mutations. This situation has changed due to the rapidly decreasing costs of whole genome (re)sequencing. DNA microarray-based mRNA expression analysis and whole genome resequencing were combined in a study on lactate transport in Saccharomyces cerevisiae. Jen1p is the only S. cerevisiae lactate transporter reported in literature. To identify alternative lactate transporters, a jen1Δ strain was evolved for growth on lactate. Results: Two independent evolution experiments yielded Jen1p-independent growth on lactate (μmax 0.14 and 0.18 h-1 for single-cell lines IMW004 and IMW005, respectively). Whereas mRNA expression analysis did not provide leads, whole-genome resequencing showed different single nucleotide changes (C755G/Leu219Val and C655G/Ala252Gly) in the acetate transporter gene ADY2. Analysis of mRNA levels and depth of coverage of DNA sequencing combined with karyotyping, gene deletions and diagnostic PCR showed that in IMW004 an isochromosome III (~475 kb), which contains two additional copies of ADY2C755G, was formed via crossover between YCLWΔ15 and YCRCΔ6. Introduction of the ADY2 alleles in a jen1 ady2 strain resulted in growth on lactate (μmax 0.14 h-1 for Ady2pLeu219Val and 0.12 h-1 for Ady2pAla252Gly). Conclusions: Whole-genome resequencing of yeast strains obtained from independent evolution experiments enabled rapid identification of a key gene that was not identified by mRNA expression analysis of the same strains. Reverse metabolic engineering showed that mutated alleles of ADY2 (C655G and C755G) encode efficient lactate transporters. The goal of the present study was to investigate whether evolutionary engineering and subsequent elucidation of the underlying mutations can lead to the identification of alternative lactate transporters in S. cerevisiae. Transport of carboxylic acids plays a key role in weak organic acids stress and the responsible membrane transporters are often poorly studied and encoded by multiple redundant genes . Additionally, import of lactate is an essential step in the reutilization of lactate produced during intense exercise in mammals [8]. In S. cerevisiae, Jen1p was previously identified as the only efficient lactate importer by generation of a UV-mutant unable to grow on lactate and subsequent functional complementation with a genomic library . The lactate uptake rate of the resulting jen1 knockout strain was close to the detection limit. Interestingly, export of lactate in engineered lactate producing S. cerevisiae is unaffected by deletion of JEN1 (our unpublished results). These observations suggest the presence of at least one alternative lactate transporter. To test this, a jen1 knockout strain was constructed and evolved for faster growth on lactate as the sole carbon and energy source. The evolved strains were subjected to a combination of mRNA expression analysis and whole genome DNA (re)sequencing to identify the relevant mutations. The resulting leads were tested for lactate transport activity via knockout studies in the evolved strains and reverse engineering of the portable genetic elements into non-evolved strains.
2011-12-21 | E-GEOD-31634 | biostudies-arrayexpress