m extorquens am1  (ATCC)

Journal: Antonie Van Leeuwenhoek

Article Title: Methylotrophic bacteria with cobalamin-dependent mutases in primary metabolism as potential strains for vitamin B 12 production

doi: 10.1007/s10482-022-01795-9

Figure Lengend Snippet: Detection of vitamin B 12 in the cell extract of M. extorquens AM1. Shown is an MRM-chromatogram of the cell extract obtained from cultivation of M. extorquens AM1 in minimal medium with 0.5% (v/v) methanol

Article Snippet: M. extorquens AM1 (DSM1338) and all strains identified in this work were obtained from DSMZ (German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany) and propagated in the media recommended by DSMZ or previously described in literature for every specific strain.

Techniques:

Journal: Antonie Van Leeuwenhoek

Article Title: Methylotrophic bacteria with cobalamin-dependent mutases in primary metabolism as potential strains for vitamin B 12 production

doi: 10.1007/s10482-022-01795-9

Figure Lengend Snippet: Comparison of vitamin B 12 amounts produced with Hyphomicrobium sp. DSM3646 and M. extorquens AM1 in the minimal medium with 0.5% methanol. The data are represented as the mean values and standard deviations of three biological replicates. Two-sample t-test was used for statistical comparison between the strains, differences considered as significant at p < 0.05 are denoted by an asterisk

Article Snippet: M. extorquens AM1 (DSM1338) and all strains identified in this work were obtained from DSMZ (German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany) and propagated in the media recommended by DSMZ or previously described in literature for every specific strain.

Techniques: Comparison, Produced

Journal: Frontiers in Microbiology

Article Title: Continuous Culture Adaptation of Methylobacterium extorquens AM1 and TK 0001 to Very High Methanol Concentrations

doi: 10.3389/fmicb.2019.01313

Figure Lengend Snippet: Continuous culture adaptation of M. extorquens TK 0001 bacteria to increasing methanol concentrations in a GM3 device. (A) Growth rate recorded for 260 days of a bacterial population cultivated on augmenting methanol concentrations. Bronze areas correspond to periods of cultivation under medium-swap regime. Black lines show the average daily generation time during cultivation periods under turbidostat regime. The methanol concentration of the culture medium applied for each evolution step (delimited by vertical gray bars) is indicated on top of the graph. Blue dots indicate time points of sampling and isolation of intermediate strains. The designation of the strain inoculated for the following adaptation step is indicated. (B) Evolutionary kinetics of adaptive growth on minimal medium 5% methanol of M. extorquens TK 0001 wildtype bacteria cultivated under medium-swap regime. A generation time of 10 h was set by the volume of the medium pulses injected at regular time intervals. The daily ratio of stressing medium pulses is plotted as a function of time. (C) Evolutionary kinetics of adaptive growth on minimal medium 7% methanol of the 5% methanol-adapted strain G4105 cultivated under medium-swap regime. A generation time of 13 h was set. The daily ratio of stressing medium pulses is plotted as a function of time.

Article Snippet: The strains M. extorquens AM1 (DSM 1338) and TK 0001 (DSM 1337) were grown at 30°C on a standard mineral (SM) medium (DSMZ n°1629) supplemented with methanol at different concentrations (v/v) as the sole carbon source.

Techniques: Bacteria, Concentration Assay, Sampling, Isolation, Injection

Journal: Frontiers in Microbiology

Article Title: Continuous Culture Adaptation of Methylobacterium extorquens AM1 and TK 0001 to Very High Methanol Concentrations

doi: 10.3389/fmicb.2019.01313

Figure Lengend Snippet: Growth phenotype of M. extorquens TK 0001 wildtype strain (blue bars) and methanol evolved isolates G4105 (green bars) and G4521 (gray bars). Generation time during exponential growth phase (A) and maximal OD 600 nm as an estimate of biomass yield (B) were determined after cultivation on minimal medium supplemented with 0.25 or 1% methanol (v/v) in shaking flasks. Each result is the mean of triplicates; ± standard deviation is indicated.

Article Snippet: The strains M. extorquens AM1 (DSM 1338) and TK 0001 (DSM 1337) were grown at 30°C on a standard mineral (SM) medium (DSMZ n°1629) supplemented with methanol at different concentrations (v/v) as the sole carbon source.

Techniques: Standard Deviation

Journal: Frontiers in Microbiology

Article Title: Continuous Culture Adaptation of Methylobacterium extorquens AM1 and TK 0001 to Very High Methanol Concentrations

doi: 10.3389/fmicb.2019.01313

Figure Lengend Snippet: Plate reader growth profiles of M. extorquens TK 0001 wildtype strain and evolved isolates (A) and of M. extorquens AM1 wildtype strain and evolved isolates (B) on SM media supplemented with 5, 7, and 8% methanol (v/v). Growth experiments were performed in triplicates; measure deviations between samples were less than 5%.

Article Snippet: The strains M. extorquens AM1 (DSM 1338) and TK 0001 (DSM 1337) were grown at 30°C on a standard mineral (SM) medium (DSMZ n°1629) supplemented with methanol at different concentrations (v/v) as the sole carbon source.

Techniques:

Journal: Frontiers in Microbiology

Article Title: Continuous Culture Adaptation of Methylobacterium extorquens AM1 and TK 0001 to Very High Methanol Concentrations

doi: 10.3389/fmicb.2019.01313

Figure Lengend Snippet: Genetic differences in isolates sampled at various time points of the adaptation of M. extorquens TK 0001 to high methanol concentrations.

Article Snippet: The strains M. extorquens AM1 (DSM 1338) and TK 0001 (DSM 1337) were grown at 30°C on a standard mineral (SM) medium (DSMZ n°1629) supplemented with methanol at different concentrations (v/v) as the sole carbon source.

Techniques:

Journal: Frontiers in Microbiology

Article Title: Continuous Culture Adaptation of Methylobacterium extorquens AM1 and TK 0001 to Very High Methanol Concentrations

doi: 10.3389/fmicb.2019.01313

Figure Lengend Snippet: Continuous culture adaptation of M. extorquens AM1 bacteria to increasing methanol concentrations in a GM3 device. (A) Growth rate recorded for 290 days of a bacterial population cultivated on augmenting methanol concentrations. Bronze areas correspond to periods of cultivation under medium-swap regime. Black lines show the average daily generation times during cultivation periods under turbidostat regime. The methanol concentration of the culture medium applied for each evolution step (delimited by vertical gray bars) is indicated on top of the graph. Blue dots indicate time points of sampling and isolation of adaptation intermediate strains for physiology and genomic analysis. (B) Evolutionary kinetics of adaptive growth on minimal medium supplemented with 5% methanol of M. extorquens AM1 wildtype bacteria cultivated under medium-swap regime. A generation time of 8 h was set by the volume of the medium pulses injected at regular time intervals. The daily ratio of stressing medium pulses is plotted as a function of time. (C) Evolutionary kinetics of adaptive growth on minimal medium supplemented with 7% methanol of the 5% methanol-adapted AM1 cell population cultivated under medium-swap regime. A generation time of 7 h 30 was set. The daily ratio of stressing medium pulses is plotted as a function of time.

Article Snippet: The strains M. extorquens AM1 (DSM 1338) and TK 0001 (DSM 1337) were grown at 30°C on a standard mineral (SM) medium (DSMZ n°1629) supplemented with methanol at different concentrations (v/v) as the sole carbon source.

Techniques: Bacteria, Concentration Assay, Sampling, Isolation, Injection

Journal: Frontiers in Microbiology

Article Title: Continuous Culture Adaptation of Methylobacterium extorquens AM1 and TK 0001 to Very High Methanol Concentrations

doi: 10.3389/fmicb.2019.01313

Figure Lengend Snippet: Genetic differences in isolates sampled at various time points of the adaptation of M. extorquens AM1 to high methanol concentrations.

Article Snippet: The strains M. extorquens AM1 (DSM 1338) and TK 0001 (DSM 1337) were grown at 30°C on a standard mineral (SM) medium (DSMZ n°1629) supplemented with methanol at different concentrations (v/v) as the sole carbon source.

Techniques: Membrane

Journal: Frontiers in Microbiology

Article Title: Continuous Culture Adaptation of Methylobacterium extorquens AM1 and TK 0001 to Very High Methanol Concentrations

doi: 10.3389/fmicb.2019.01313

Figure Lengend Snippet: Mutations in MetY identified in the isolates analyzed at each step of the adaptation to increasing concentrations of methanol of both M. extorquens strains TK 0001 and AM1.

Article Snippet: The strains M. extorquens AM1 (DSM 1338) and TK 0001 (DSM 1337) were grown at 30°C on a standard mineral (SM) medium (DSMZ n°1629) supplemented with methanol at different concentrations (v/v) as the sole carbon source.

Techniques:

Journal: Frontiers in Microbiology

Article Title: Continuous Culture Adaptation of Methylobacterium extorquens AM1 and TK 0001 to Very High Methanol Concentrations

doi: 10.3389/fmicb.2019.01313

Figure Lengend Snippet: Kinetic parameters of metY -encoded wildtype O -acetyl- L -homoserine sulfhydrylase of M. extorquens TK 0001.

Article Snippet: The strains M. extorquens AM1 (DSM 1338) and TK 0001 (DSM 1337) were grown at 30°C on a standard mineral (SM) medium (DSMZ n°1629) supplemented with methanol at different concentrations (v/v) as the sole carbon source.

Techniques:

Journal: Frontiers in Microbiology

Article Title: Continuous Culture Adaptation of Methylobacterium extorquens AM1 and TK 0001 to Very High Methanol Concentrations

doi: 10.3389/fmicb.2019.01313

Figure Lengend Snippet: Growth of M. extorquens methanol adapted strain G4105 transformed with the empty vector pTE102 (light green bars) or the plasmid pTE102_ metY (dark green bars). Generation time (A) and maximal OD 600 nm as an estimate of biomass yield (B) were determined after cultivation on minimal medium supplemented with 0.25, 1, or 5% methanol (v/v). Each result is the mean of triplicates; ± standard deviation is indicated.

Article Snippet: The strains M. extorquens AM1 (DSM 1338) and TK 0001 (DSM 1337) were grown at 30°C on a standard mineral (SM) medium (DSMZ n°1629) supplemented with methanol at different concentrations (v/v) as the sole carbon source.

Techniques: Transformation Assay, Plasmid Preparation, Standard Deviation

Journal: Frontiers in Microbiology

Article Title: Continuous Culture Adaptation of Methylobacterium extorquens AM1 and TK 0001 to Very High Methanol Concentrations

doi: 10.3389/fmicb.2019.01313

Figure Lengend Snippet: Comparative analysis of the transcriptomes of M. extorquens TK 0001 wildtype strain and G4105 methanol-adapted strain upon exposure to 1 and 5% methanol. Differentially expressed genes (DEGs) [log 2 (fold change) >2 or <2] were categorized based on eggNOG functional classification. Red bars represent the number of upregulated genes and green bars the number of downregulated genes in each eggNOG class. (A) Number of DEGs in TK 0001 cells exposed to 5% versus TK 0001 cells exposed to 1% methanol for 5 min. (B) Number of DEGs in G4105 cells exposed to 5% versus G4105 cells exposed to 1% methanol for 5 min. (C) Number of DEGs in TK 0001 cells exposed to 5% methanol for 3 h versus TK 0001 cells exposed to 5% methanol for 5 min. (D) Number of DEGs in G4105 cells exposed to 5% methanol for 3 h versus G4105 cells exposed to 5% methanol for 5 min. (E) Number of DEGs in G4105 versus TK 0001 cells, similarly exposed to 1% methanol for 5 min. (F) Number of DEGs in G4105 versus TK 0001 cells, similarly exposed to 5% methanol for 5 min. (G) Number of DEGs in G4105 versus TK 0001 cells, similarly exposed to 1% methanol for 3 h. (H) Number of DEGs in G4105 versus TK 0001 cells, similarly exposed to 5% methanol for 3 h.

Article Snippet: The strains M. extorquens AM1 (DSM 1338) and TK 0001 (DSM 1337) were grown at 30°C on a standard mineral (SM) medium (DSMZ n°1629) supplemented with methanol at different concentrations (v/v) as the sole carbon source.

Techniques: Functional Assay

Journal: Frontiers in Microbiology

Article Title: Continuous Culture Adaptation of Methylobacterium extorquens AM1 and TK 0001 to Very High Methanol Concentrations

doi: 10.3389/fmicb.2019.01313

Figure Lengend Snippet: Lactate intracellular concentration (mean ± SEM) formed by G4605 ( n = 23), G4836 ( n = 23), G4891 ( n = 23), and G4892 ( n = 23). Data were normalized for 2.5 ml cell cultures at an OD 600 nm = 1. A non-parametric Kruskal–Wallis test followed by Dunn test was used to determine difference between groups. Bars not sharing the same letter are statistically different ( p < 0.0083). Cellular concentrations were calculated assuming the same cell internal volume 2.310 –9 μl for M. extorquens as for E. coli .

Article Snippet: The strains M. extorquens AM1 (DSM 1338) and TK 0001 (DSM 1337) were grown at 30°C on a standard mineral (SM) medium (DSMZ n°1629) supplemented with methanol at different concentrations (v/v) as the sole carbon source.

Techniques: Concentration Assay