Journal: Microbial Biotechnology

Article Title: De novo biosynthesis of garbanzol and fustin in Streptomyces albus based on a potential flavanone 3‐hydroxylase with 2‐hydroxylase side activity

doi: 10.1111/1751-7915.13874

Figure Lengend Snippet: Designed gene clusters for heterologous expression of garbanzol (A) and fustin (B) in S. albus . Both DNA constructions were finally subcloned as a Bgl II‐ Bam HI fragment into the pIAGO plasmid. Blocked restriction enzymes sites (after DNA polymerase I Klenow fragment treatment during subcloning) are represented in brackets.

Article Snippet: Authentic standards for LC‐HRESIMS quantification and molecule identification through LC‐HRESIMS and LC‐UV/vis were purchased from different suppliers: p ‐coumaric acid (Sigma Aldrich, USA), naringenin (Sigma Aldrich, USA), dihydrokaempferol (Sigma Aldrich, USA), kaempferol (Cayman Chemical, USA), apigenin (Extrasynthese, Genay, France), 2‐hydroxynaringenin (Ambinter, Orléans, France), isoliquiritigenin (Sigma Aldrich, USA), liquiritigenin (Tocris Bioscience, Bristol, UK), (2 R ,3 R )‐garbanzol (BioBioPha, Kunming, Yunnan, China), (2 S ,3 S )‐garbanzol (AnalytiCon Discovery, Potsdam, Germany), resokaempferol (Extrasynthese), fustin (Biosynth Carbosynth, Compton, UK), butin (ChemFaces, Wuhan, China), 7,4′‐dihydroxyflavone (Extrasynthese, France), quercetin (Cayman Chemical) and dihydroquercetin (Sigma Aldrich).

Techniques: Expressing, Plasmid Preparation, Subcloning

Journal: Microbial Biotechnology

Article Title: De novo biosynthesis of garbanzol and fustin in Streptomyces albus based on a potential flavanone 3‐hydroxylase with 2‐hydroxylase side activity

doi: 10.1111/1751-7915.13874

Figure Lengend Snippet: EICs at m / z 271.06118 ± 0.005 [M‐H] − of S. albus pGR and S. albus pDF crude extract (A) and commercial flavonoids standards of NRG (naringenin) and GRB (garbanzol) (B). Peak 1 corresponds to 2‐hydroxyliquiritigenin (probably two isomers based on the stereochemistry of the 2‐hydroxyl group), peak 2 corresponds to the two commercial garbanzol enantiomers, peaks 3 and 4 probably correspond to different tautomeric forms of licodione (tentative identification) and peak 6 corresponds to naringenin. GRB, garbanzol; NRG, naringenin.

Article Snippet: Authentic standards for LC‐HRESIMS quantification and molecule identification through LC‐HRESIMS and LC‐UV/vis were purchased from different suppliers: p ‐coumaric acid (Sigma Aldrich, USA), naringenin (Sigma Aldrich, USA), dihydrokaempferol (Sigma Aldrich, USA), kaempferol (Cayman Chemical, USA), apigenin (Extrasynthese, Genay, France), 2‐hydroxynaringenin (Ambinter, Orléans, France), isoliquiritigenin (Sigma Aldrich, USA), liquiritigenin (Tocris Bioscience, Bristol, UK), (2 R ,3 R )‐garbanzol (BioBioPha, Kunming, Yunnan, China), (2 S ,3 S )‐garbanzol (AnalytiCon Discovery, Potsdam, Germany), resokaempferol (Extrasynthese), fustin (Biosynth Carbosynth, Compton, UK), butin (ChemFaces, Wuhan, China), 7,4′‐dihydroxyflavone (Extrasynthese, France), quercetin (Cayman Chemical) and dihydroquercetin (Sigma Aldrich).

Techniques:

Journal: Microbial Biotechnology

Article Title: De novo biosynthesis of garbanzol and fustin in Streptomyces albus based on a potential flavanone 3‐hydroxylase with 2‐hydroxylase side activity

doi: 10.1111/1751-7915.13874

Figure Lengend Snippet: Cultivation data (dry cell weight) of S. albus pGR (red) and negative control S. albus pIAGO (green) grown on R5A medium sampled every 24 h over 168 h. Garbanzol titres in μg l −1 have been overlayed (dashed line). The data are expressed as mean value ± standard error of mean (SEM). * indicate statistically significant differences between both strains ( S. albus pGR and negative control S. albus pIAGO) after two‐way ANOVA.

Article Snippet: Authentic standards for LC‐HRESIMS quantification and molecule identification through LC‐HRESIMS and LC‐UV/vis were purchased from different suppliers: p ‐coumaric acid (Sigma Aldrich, USA), naringenin (Sigma Aldrich, USA), dihydrokaempferol (Sigma Aldrich, USA), kaempferol (Cayman Chemical, USA), apigenin (Extrasynthese, Genay, France), 2‐hydroxynaringenin (Ambinter, Orléans, France), isoliquiritigenin (Sigma Aldrich, USA), liquiritigenin (Tocris Bioscience, Bristol, UK), (2 R ,3 R )‐garbanzol (BioBioPha, Kunming, Yunnan, China), (2 S ,3 S )‐garbanzol (AnalytiCon Discovery, Potsdam, Germany), resokaempferol (Extrasynthese), fustin (Biosynth Carbosynth, Compton, UK), butin (ChemFaces, Wuhan, China), 7,4′‐dihydroxyflavone (Extrasynthese, France), quercetin (Cayman Chemical) and dihydroquercetin (Sigma Aldrich).

Techniques: Negative Control

Journal: Microbial Biotechnology

Article Title: De novo biosynthesis of garbanzol and fustin in Streptomyces albus based on a potential flavanone 3‐hydroxylase with 2‐hydroxylase side activity

doi: 10.1111/1751-7915.13874

Figure Lengend Snippet: HPLC‐HRESIMS/MS analysis of authentic garbanzol and purified 2‐hydroxyliquitigenin. According to the nomenclature of Yang et al. each fragment is denoted by the combined use of i,j A − or i,j B − . A and B represent the flavonoid intact ring, and the superscript on the left indicates the broken bounds of the deprotonated molecule (Yang et al ., ). A, Proposed retrocyclization cleavages of the C ring. B, MS 2 fragmentation products for the parent ion m / z 271.0535 [M‐H] − . C, MS 2 fragmentation products for the in‐source‐formed species m / z 243.0591 [M‐H] − . Product ions resulting from losses of H 2 O, CO and CO 2 have also been included.

Article Snippet: Authentic standards for LC‐HRESIMS quantification and molecule identification through LC‐HRESIMS and LC‐UV/vis were purchased from different suppliers: p ‐coumaric acid (Sigma Aldrich, USA), naringenin (Sigma Aldrich, USA), dihydrokaempferol (Sigma Aldrich, USA), kaempferol (Cayman Chemical, USA), apigenin (Extrasynthese, Genay, France), 2‐hydroxynaringenin (Ambinter, Orléans, France), isoliquiritigenin (Sigma Aldrich, USA), liquiritigenin (Tocris Bioscience, Bristol, UK), (2 R ,3 R )‐garbanzol (BioBioPha, Kunming, Yunnan, China), (2 S ,3 S )‐garbanzol (AnalytiCon Discovery, Potsdam, Germany), resokaempferol (Extrasynthese), fustin (Biosynth Carbosynth, Compton, UK), butin (ChemFaces, Wuhan, China), 7,4′‐dihydroxyflavone (Extrasynthese, France), quercetin (Cayman Chemical) and dihydroquercetin (Sigma Aldrich).

Techniques: Purification

Journal: Microbial Biotechnology

Article Title: De novo biosynthesis of garbanzol and fustin in Streptomyces albus based on a potential flavanone 3‐hydroxylase with 2‐hydroxylase side activity

doi: 10.1111/1751-7915.13874

Figure Lengend Snippet: Designed gene clusters for heterologous expression of garbanzol (A) and fustin (B) in S. albus . Both DNA constructions were finally subcloned as a Bgl II‐ Bam HI fragment into the pIAGO plasmid. Blocked restriction enzymes sites (after DNA polymerase I Klenow fragment treatment during subcloning) are represented in brackets.

Article Snippet: Authentic standards for LC‐HRESIMS quantification and molecule identification through LC‐HRESIMS and LC‐UV/vis were purchased from different suppliers: p ‐coumaric acid (Sigma Aldrich, USA), naringenin (Sigma Aldrich, USA), dihydrokaempferol (Sigma Aldrich, USA), kaempferol (Cayman Chemical, USA), apigenin (Extrasynthese, Genay, France), 2‐hydroxynaringenin (Ambinter, Orléans, France), isoliquiritigenin (Sigma Aldrich, USA), liquiritigenin (Tocris Bioscience, Bristol, UK), (2 R ,3 R )‐garbanzol (BioBioPha, Kunming, Yunnan, China), (2 S ,3 S )‐garbanzol (AnalytiCon Discovery, Potsdam, Germany), resokaempferol (Extrasynthese), fustin (Biosynth Carbosynth, Compton, UK), butin (ChemFaces, Wuhan, China), 7,4′‐dihydroxyflavone (Extrasynthese, France), quercetin (Cayman Chemical) and dihydroquercetin (Sigma Aldrich).

Techniques: Expressing, Plasmid Preparation, Subcloning

Journal: Microbial Biotechnology

Article Title: De novo biosynthesis of garbanzol and fustin in Streptomyces albus based on a potential flavanone 3‐hydroxylase with 2‐hydroxylase side activity

doi: 10.1111/1751-7915.13874

Figure Lengend Snippet: EICs at m / z 271.06118 ± 0.005 [M‐H] − of S. albus pGR and S. albus pDF crude extract (A) and commercial flavonoids standards of NRG (naringenin) and GRB (garbanzol) (B). Peak 1 corresponds to 2‐hydroxyliquiritigenin (probably two isomers based on the stereochemistry of the 2‐hydroxyl group), peak 2 corresponds to the two commercial garbanzol enantiomers, peaks 3 and 4 probably correspond to different tautomeric forms of licodione (tentative identification) and peak 6 corresponds to naringenin. GRB, garbanzol; NRG, naringenin.

Article Snippet: Authentic standards for LC‐HRESIMS quantification and molecule identification through LC‐HRESIMS and LC‐UV/vis were purchased from different suppliers: p ‐coumaric acid (Sigma Aldrich, USA), naringenin (Sigma Aldrich, USA), dihydrokaempferol (Sigma Aldrich, USA), kaempferol (Cayman Chemical, USA), apigenin (Extrasynthese, Genay, France), 2‐hydroxynaringenin (Ambinter, Orléans, France), isoliquiritigenin (Sigma Aldrich, USA), liquiritigenin (Tocris Bioscience, Bristol, UK), (2 R ,3 R )‐garbanzol (BioBioPha, Kunming, Yunnan, China), (2 S ,3 S )‐garbanzol (AnalytiCon Discovery, Potsdam, Germany), resokaempferol (Extrasynthese), fustin (Biosynth Carbosynth, Compton, UK), butin (ChemFaces, Wuhan, China), 7,4′‐dihydroxyflavone (Extrasynthese, France), quercetin (Cayman Chemical) and dihydroquercetin (Sigma Aldrich).

Techniques:

Journal: Microbial Biotechnology

Article Title: De novo biosynthesis of garbanzol and fustin in Streptomyces albus based on a potential flavanone 3‐hydroxylase with 2‐hydroxylase side activity

doi: 10.1111/1751-7915.13874

Figure Lengend Snippet: Cultivation data (dry cell weight) of S. albus pGR (red) and negative control S. albus pIAGO (green) grown on R5A medium sampled every 24 h over 168 h. Garbanzol titres in μg l −1 have been overlayed (dashed line). The data are expressed as mean value ± standard error of mean (SEM). * indicate statistically significant differences between both strains ( S. albus pGR and negative control S. albus pIAGO) after two‐way ANOVA.

Article Snippet: Authentic standards for LC‐HRESIMS quantification and molecule identification through LC‐HRESIMS and LC‐UV/vis were purchased from different suppliers: p ‐coumaric acid (Sigma Aldrich, USA), naringenin (Sigma Aldrich, USA), dihydrokaempferol (Sigma Aldrich, USA), kaempferol (Cayman Chemical, USA), apigenin (Extrasynthese, Genay, France), 2‐hydroxynaringenin (Ambinter, Orléans, France), isoliquiritigenin (Sigma Aldrich, USA), liquiritigenin (Tocris Bioscience, Bristol, UK), (2 R ,3 R )‐garbanzol (BioBioPha, Kunming, Yunnan, China), (2 S ,3 S )‐garbanzol (AnalytiCon Discovery, Potsdam, Germany), resokaempferol (Extrasynthese), fustin (Biosynth Carbosynth, Compton, UK), butin (ChemFaces, Wuhan, China), 7,4′‐dihydroxyflavone (Extrasynthese, France), quercetin (Cayman Chemical) and dihydroquercetin (Sigma Aldrich).

Techniques: Negative Control

Journal: Microbial Biotechnology

Article Title: De novo biosynthesis of garbanzol and fustin in Streptomyces albus based on a potential flavanone 3‐hydroxylase with 2‐hydroxylase side activity

doi: 10.1111/1751-7915.13874

Figure Lengend Snippet: HPLC‐HRESIMS/MS analysis of authentic garbanzol and purified 2‐hydroxyliquitigenin. According to the nomenclature of Yang et al. each fragment is denoted by the combined use of i,j A − or i,j B − . A and B represent the flavonoid intact ring, and the superscript on the left indicates the broken bounds of the deprotonated molecule (Yang et al ., ). A, Proposed retrocyclization cleavages of the C ring. B, MS 2 fragmentation products for the parent ion m / z 271.0535 [M‐H] − . C, MS 2 fragmentation products for the in‐source‐formed species m / z 243.0591 [M‐H] − . Product ions resulting from losses of H 2 O, CO and CO 2 have also been included.

Article Snippet: Authentic standards for LC‐HRESIMS quantification and molecule identification through LC‐HRESIMS and LC‐UV/vis were purchased from different suppliers: p ‐coumaric acid (Sigma Aldrich, USA), naringenin (Sigma Aldrich, USA), dihydrokaempferol (Sigma Aldrich, USA), kaempferol (Cayman Chemical, USA), apigenin (Extrasynthese, Genay, France), 2‐hydroxynaringenin (Ambinter, Orléans, France), isoliquiritigenin (Sigma Aldrich, USA), liquiritigenin (Tocris Bioscience, Bristol, UK), (2 R ,3 R )‐garbanzol (BioBioPha, Kunming, Yunnan, China), (2 S ,3 S )‐garbanzol (AnalytiCon Discovery, Potsdam, Germany), resokaempferol (Extrasynthese), fustin (Biosynth Carbosynth, Compton, UK), butin (ChemFaces, Wuhan, China), 7,4′‐dihydroxyflavone (Extrasynthese, France), quercetin (Cayman Chemical) and dihydroquercetin (Sigma Aldrich).

Techniques: Purification