hplc ultimate 3000  (Thermo Fisher)

Journal: Metabolites

Article Title: HR-LCMS/MS-Based Dereplication of Plant-Derived Autophagy Inducers Revealed Astragalus dasyanthus as a New Glabrol Producer

doi: 10.3390/metabo16050311

Figure Lengend Snippet: Identification of the active compound in Saponaria officinalis . ( a ) Western blot analysis of the ability of HPLC fractions of the ethanolic extract of Saponaria officinalis to induce autophagy (LC3 I/II staining: the panel shows the fragment containing the active fraction; the full image is presented in ; densitometric analysis of the LC3 II/LC3 I ratio is shown under the staining result; the fraction sent for further analysis is marked in bold). ( b ) MS/MS comparison of the observed ion in positive-ion mode (top graph, m / z 595.156) with the deposited GNPS reference spectrum for saponarin [M + H] + (bottom graph, GNPS LIBRARY: accession: CCMSLIB00010125140, m / z 595.1660). ( c ) Chemical structure of saponarin.

Article Snippet: The analysis was performed using an UltiMate 3000 Acclaim RSLC HPLC system (Fisher Scientific, Waltham, MA, USA) with a 120 C18 2.2 um 2.1 × 100 mm column and a gradient of acetonitrile (0.1% FA) in water (0.1% TFA) of 5–95% for 10 min, while connected to a Bruker maXis II 4 G ETD (Bruker, Billerica, MA, USA).

Techniques: Western Blot, Staining, Tandem Mass Spectroscopy, Comparison

Journal: Metabolites

Article Title: HR-LCMS/MS-Based Dereplication of Plant-Derived Autophagy Inducers Revealed Astragalus dasyanthus as a New Glabrol Producer

doi: 10.3390/metabo16050311

Figure Lengend Snippet: Identification of the active compounds in Veronica officinalis . ( a ) Western blot analysis of the ability of HPLC fractions of the ethanolic extract of Veronica officinalis to induce autophagy (LC3 I/II staining: the panel shows the fragment containing the active fraction; the full image is presented in , and densitometric analysis of the LC3 II/LC3 I ratio is shown under the staining result; the fraction sent for further analysis is marked in bold). ( b ) MS/MS comparison of the observed ions in positive-ion mode ( m / z 447.091) and negative-ion mode ( m / z 445.076) with the deposited GNPS reference spectra for baicalin [M + H] + and [M − H] − (GNPS-LIBRARY: accession: CCMSLIB00013029693, m / z 447.092; GNPS-LIBRARY: accession: CCMSLIB00012855511, m / z 445.078, respectively). ( c ) Chemical structure of baicalin.

Article Snippet: The analysis was performed using an UltiMate 3000 Acclaim RSLC HPLC system (Fisher Scientific, Waltham, MA, USA) with a 120 C18 2.2 um 2.1 × 100 mm column and a gradient of acetonitrile (0.1% FA) in water (0.1% TFA) of 5–95% for 10 min, while connected to a Bruker maXis II 4 G ETD (Bruker, Billerica, MA, USA).

Techniques: Western Blot, Staining, Tandem Mass Spectroscopy, Comparison

Journal: Metabolites

Article Title: HR-LCMS/MS-Based Dereplication of Plant-Derived Autophagy Inducers Revealed Astragalus dasyanthus as a New Glabrol Producer

doi: 10.3390/metabo16050311

Figure Lengend Snippet: Identification of the active compounds in Astragalus dasyanthus . ( a ) Western blot analysis of the ability of HPLC fractions of the ethanolic extract of Astragalus dasyanthus to induce autophagy (LC3 I/II staining: the panel shows the fragment containing the active fraction; the full image is presented in ; the fraction sent for further analysis is marked in bold). ( b ) MS/MS comparison of the observed ion in negative-ion mode ( m / z 391.1910) with the reference spectrum for glabrol [M − H] − (GNPS-LIBRARY; accession: CCMSLIB00005749603, m / z 393.19). ( c ) Chemical structure of glabrol.

Article Snippet: The analysis was performed using an UltiMate 3000 Acclaim RSLC HPLC system (Fisher Scientific, Waltham, MA, USA) with a 120 C18 2.2 um 2.1 × 100 mm column and a gradient of acetonitrile (0.1% FA) in water (0.1% TFA) of 5–95% for 10 min, while connected to a Bruker maXis II 4 G ETD (Bruker, Billerica, MA, USA).

Techniques: Western Blot, Staining, Tandem Mass Spectroscopy, Comparison

Journal: Synthetic and Systems Biotechnology

Article Title: Recombinant production of amaranthin and other betalain variants with yeast cell factories

doi: 10.1016/j.synbio.2025.05.008

Figure Lengend Snippet: Screening of glucuronosyltransferases in betanin-producing S. cerevisiae . a HPLC-UV chromatogram, comparing the total fraction of the parent strain ST12160 with the strains expressing Ah AmaSy1, Cq AmaSy1 or Cc AmaSy1 and the plant extract from B. glabra . A compound with an RT of 5.4 min, also present in the plant, was detected in the strains expressing Ah AmaSy1 and Cc AmaSy1. The upward arrow (↑) indicates overexpression of the gene listed after the symbol. b Extracted-ion chromatogram (XIC) and the corresponding MS 2 spectrum from the LC-MS analysis revealed that this new compound with m / z = 713.2039 is bougainvillein-r I. The product ion at m / z = 389.09, characteristic for all betacyanins, corresponds to betanidin, the product ion at m / z = 343.09 corresponds to monodecarboxylated betanidin.

Article Snippet: To quantify the betalains produced by the yeast cultures, the samples were analysed via high-performance liquid chromatography (HPLC) using a Dionex Ultimate 3000 HPLC system (Thermo Fisher Scientific, US).

Techniques: Expressing, Plant Extract, Over Expression, Liquid Chromatography with Mass Spectroscopy

Journal: Synthetic and Systems Biotechnology

Article Title: Recombinant production of amaranthin and other betalain variants with yeast cell factories

doi: 10.1016/j.synbio.2025.05.008

Figure Lengend Snippet: UDP-glucuronic acid synthesis in S. cerevisiae enables the production of amaranthin by glucuronosyltransferases . a Depending on the available sugar donor, betanin is glycosylated to bougainvillein-r I or amaranthin by the glucuronosyltransferases. At UGD1 converts UDP-glucose to UDP-glucuronic acid. b UDP-glucose and UDP-glucuronic acid concentrations in betalain producing S. cerevisiae without (ST12160) and with expression of a heterologous UDP-glucose dehydrogenase ( At UGD1). c HPLC analysis detects a new compound in all three strains expressing At UGD1 in combination with a glucuronosyltransferase that is also present in the plant extract from A. cruentus . LC-MS analysis confirms that this compound is amaranthin. d Betalain production in strains with or without At UGD1, quantified by HPLC. Concentrations of C15 stereoisomers were combined. Mean values of biological triplicates (±SD) are shown. “X” indicates the integration of At UGD1. e Betalain-producing S. cerevisiae strains on agar plate. At UGD1: UDP-glucose dehydrogenase ( A. thaliana ), Ah AmaSy1: Glucuronosyltransferase ( Amaranthus hypochondriacus ), Cq AmaSy1: Glucuronosyltransferase ( Chenopodium quinoa ), Cc AmaSy1: Glucuronosyltransferase ( Celosia argentea var. cristata ), Mj DOD: 4,5-extradiol dioxyge nase ( Mirabilis jalapa ), Bv TYH: tyrosine hydroxylase ( Beta vulgaris ), Bv GT2: UDP-glycosyltransferase ( Beta vulgaris ). The upward arrow (↑) indicates overexpression of the gene listed after the symbol.

Article Snippet: To quantify the betalains produced by the yeast cultures, the samples were analysed via high-performance liquid chromatography (HPLC) using a Dionex Ultimate 3000 HPLC system (Thermo Fisher Scientific, US).

Techniques: Expressing, Plant Extract, Liquid Chromatography with Mass Spectroscopy, Over Expression

Journal: Synthetic and Systems Biotechnology

Article Title: Recombinant production of amaranthin and other betalain variants with yeast cell factories

doi: 10.1016/j.synbio.2025.05.008

Figure Lengend Snippet: Expression of glucuronosyltransferases in betanin-producing Y. lipolytica . ST12603 and the GlcAT-expressing strains were cultivated in 24-well plates for 48 h. a Intra- and extracellular betalain concentrations, determined by HPLC. The corresponding 10x diluted samples are shown below the graph. Concentrations of the C15 stereoisomers were combined. Mean values of biological triplicates (±SD) are shown. b UDP-glucose and UDP-glucuronic acid concentrations in the yeast strains were measured by LC-MS. The upward arrow (↑) indicates overexpression of the gene listed after the symbol.

Article Snippet: To quantify the betalains produced by the yeast cultures, the samples were analysed via high-performance liquid chromatography (HPLC) using a Dionex Ultimate 3000 HPLC system (Thermo Fisher Scientific, US).

Techniques: Expressing, Liquid Chromatography with Mass Spectroscopy, Over Expression

Journal: Synthetic and Systems Biotechnology

Article Title: Recombinant production of amaranthin and other betalain variants with yeast cell factories

doi: 10.1016/j.synbio.2025.05.008

Figure Lengend Snippet: Celoscristatin production in S. cerevisiae and Y. lipolytica . a To synthesise malonylated amaranthin from betanin, a glucuronosyltransferase ( Ah / Cq / Cc AmaSy1) and a malonyltransferase (e.g. Hp BAHD3) are required. b HPLC chromatogram (540 nm) of the extracellular fraction of amaranthin-producing S. cerevisiae strains, additionally expressing the acyltransferase Hp BAHD3. A new compound was detected in all three strains, absent in the reference strains. Y-axis: 10 – 120 mAU. The upward arrow (↑) indicates overexpression of the gene listed after the symbol. c Expression of Hp BAHD3 and Cc AmaSy1 in betanin-producing Y. lipolytica led to the formation of the same compound. From LC-MS analysis, this new compound was identified as celoscristatin (6′-O-malonyl-amaranthin) . d Phyllocactin- (ST14103), amaranthin- (ST14102) and celoscristatin- (ST14760) producing Y. lipolytica strains were cultivated in MM and the extracellular and intracellular production analysed by HPLC. Peak areas (mAU∗min) were compared. Mean values of biological triplicates (±SD) are shown. e UV–vis spectrum of 10x diluted extracellular and total samples. The corresponding 2x and 10x diluted samples of ST14760 are shown.

Article Snippet: To quantify the betalains produced by the yeast cultures, the samples were analysed via high-performance liquid chromatography (HPLC) using a Dionex Ultimate 3000 HPLC system (Thermo Fisher Scientific, US).

Techniques: Expressing, Over Expression, Liquid Chromatography with Mass Spectroscopy