caption a7 strain plasmid relevant genotype  (ATCC)

Journal: Molecular microbiology

Article Title: Fatty acid activation and utilization by Alistipes finegoldii , a representative Bacteroidetes resident of the human gut microbiome

doi: 10.1111/mmi.14445

Figure Lengend Snippet: Predicted lipid metabolism genes in A. finegoldii Gene identifications were made using tBLASTn to query Uniprot sequences against A. finegoldii DSM 17242 taxid: 679935.

Article Snippet: The A 600 in the 16×100 mm borosilicate glass tubes were measured using a Biochrom WPA CO8000 cell density meter. table ft1 table-wrap mode="anchored" t5 caption a7 Strain Relevant Genotype Source A. finegoldii Strain DSM 17242 (DSMZ Collection) ( Rautio et al., 2003 ) E. coli SM2-1 plsC101 (Ts) metCl6S :: Tn10 ( Coleman, 1990 ) E. coli LCH30 aas-1 fadD88 metB1 relA1 spoT1 ( Hsu et al., 1991 ) Plasmid Description Source pPJ131 E. coli expression vector ( Lu et al., 2006 ) p Ec PlsC E. coli plsC in pPJ131 ( Robertson et al., 2017 ) p Ct PlsC Chlamydia trachomatis plsC in pPJ131 ( Robertson et al., 2017 ) p Af PlsC A. finegoldii plsC ( Alfi_2876 ) in pPJ131 This study p Ec FadD E. coli fadD in pPJ131 ( Yao et al., 2016 ) p Ng Aas Neisseria gonorrhoeae aas in pPJ131 ( Yao et al., 2016 ) p Af Aas1 A. finegoldii aas1 ( Alfi_2635 ) in pPJ131 This study p Af 3181 A. finegoldii Alfi_3181 in pPJ131 This study p Af Aas2 A. finegoldii aas2 ( Alfi_0371 ) in pPJ131 This study pET_ Af Aas1 A. finegoldii aas1 ( Alfi_2635 ) in pET This study pET_ Af Aas2 A. finegoldii aas2 ( Alfi_0371 ) in pET This study Open in a separate window Bacterial strains and plasmids The minimum inhibitory concentration for cerulenin with or without exogenous fatty acid supplementation against A. finegoldii was determined using a modified broth microdilution method ( Parsons et al., 2011 ).

Techniques:

Journal: Molecular microbiology

Article Title: Fatty acid activation and utilization by Alistipes finegoldii , a representative Bacteroidetes resident of the human gut microbiome

doi: 10.1111/mmi.14445

Figure Lengend Snippet: A. finegoldii was grown in three different media, the lipids extracted and the PE molecular species profiled by mass spectrometry. PE molecular species containing an unsaturated fatty acid are indicated in red.

Article Snippet: The A 600 in the 16×100 mm borosilicate glass tubes were measured using a Biochrom WPA CO8000 cell density meter. table ft1 table-wrap mode="anchored" t5 caption a7 Strain Relevant Genotype Source A. finegoldii Strain DSM 17242 (DSMZ Collection) ( Rautio et al., 2003 ) E. coli SM2-1 plsC101 (Ts) metCl6S :: Tn10 ( Coleman, 1990 ) E. coli LCH30 aas-1 fadD88 metB1 relA1 spoT1 ( Hsu et al., 1991 ) Plasmid Description Source pPJ131 E. coli expression vector ( Lu et al., 2006 ) p Ec PlsC E. coli plsC in pPJ131 ( Robertson et al., 2017 ) p Ct PlsC Chlamydia trachomatis plsC in pPJ131 ( Robertson et al., 2017 ) p Af PlsC A. finegoldii plsC ( Alfi_2876 ) in pPJ131 This study p Ec FadD E. coli fadD in pPJ131 ( Yao et al., 2016 ) p Ng Aas Neisseria gonorrhoeae aas in pPJ131 ( Yao et al., 2016 ) p Af Aas1 A. finegoldii aas1 ( Alfi_2635 ) in pPJ131 This study p Af 3181 A. finegoldii Alfi_3181 in pPJ131 This study p Af Aas2 A. finegoldii aas2 ( Alfi_0371 ) in pPJ131 This study pET_ Af Aas1 A. finegoldii aas1 ( Alfi_2635 ) in pET This study pET_ Af Aas2 A. finegoldii aas2 ( Alfi_0371 ) in pET This study Open in a separate window Bacterial strains and plasmids The minimum inhibitory concentration for cerulenin with or without exogenous fatty acid supplementation against A. finegoldii was determined using a modified broth microdilution method ( Parsons et al., 2011 ).

Techniques: Mass Spectrometry

Journal: Molecular microbiology

Article Title: Fatty acid activation and utilization by Alistipes finegoldii , a representative Bacteroidetes resident of the human gut microbiome

doi: 10.1111/mmi.14445

Figure Lengend Snippet: A. The growth of A. finegoldii was monitored in either DSM Medium 104 (104, red), DSM Medium 104 excluding Tween 80 (104-T, blue), or lipid-free WCAB defined medium (WCAB, black). Growth was monitored over three days, and data is presented as the mean A600 ± S.E.M. of three independent cultures.

Article Snippet: The A 600 in the 16×100 mm borosilicate glass tubes were measured using a Biochrom WPA CO8000 cell density meter. table ft1 table-wrap mode="anchored" t5 caption a7 Strain Relevant Genotype Source A. finegoldii Strain DSM 17242 (DSMZ Collection) ( Rautio et al., 2003 ) E. coli SM2-1 plsC101 (Ts) metCl6S :: Tn10 ( Coleman, 1990 ) E. coli LCH30 aas-1 fadD88 metB1 relA1 spoT1 ( Hsu et al., 1991 ) Plasmid Description Source pPJ131 E. coli expression vector ( Lu et al., 2006 ) p Ec PlsC E. coli plsC in pPJ131 ( Robertson et al., 2017 ) p Ct PlsC Chlamydia trachomatis plsC in pPJ131 ( Robertson et al., 2017 ) p Af PlsC A. finegoldii plsC ( Alfi_2876 ) in pPJ131 This study p Ec FadD E. coli fadD in pPJ131 ( Yao et al., 2016 ) p Ng Aas Neisseria gonorrhoeae aas in pPJ131 ( Yao et al., 2016 ) p Af Aas1 A. finegoldii aas1 ( Alfi_2635 ) in pPJ131 This study p Af 3181 A. finegoldii Alfi_3181 in pPJ131 This study p Af Aas2 A. finegoldii aas2 ( Alfi_0371 ) in pPJ131 This study pET_ Af Aas1 A. finegoldii aas1 ( Alfi_2635 ) in pET This study pET_ Af Aas2 A. finegoldii aas2 ( Alfi_0371 ) in pET This study Open in a separate window Bacterial strains and plasmids The minimum inhibitory concentration for cerulenin with or without exogenous fatty acid supplementation against A. finegoldii was determined using a modified broth microdilution method ( Parsons et al., 2011 ).

Techniques:

Journal: Molecular microbiology

Article Title: Fatty acid activation and utilization by Alistipes finegoldii , a representative Bacteroidetes resident of the human gut microbiome

doi: 10.1111/mmi.14445

Figure Lengend Snippet: A, A. finegoldii was grown in the presence of a series of heavy-labeled saturated or unsaturated fatty acids. The incorporation of each of these fatty acids into PE and SL was determined by mass spectrometry. There are two possible fates for the fatty acids after their entry into the cell. The exogenous fatty acid may be directly incorporated into PE or SL, or it may enter the FASII elongation cycle and its elongation product is incorporated into PE or SL.

Article Snippet: The A 600 in the 16×100 mm borosilicate glass tubes were measured using a Biochrom WPA CO8000 cell density meter. table ft1 table-wrap mode="anchored" t5 caption a7 Strain Relevant Genotype Source A. finegoldii Strain DSM 17242 (DSMZ Collection) ( Rautio et al., 2003 ) E. coli SM2-1 plsC101 (Ts) metCl6S :: Tn10 ( Coleman, 1990 ) E. coli LCH30 aas-1 fadD88 metB1 relA1 spoT1 ( Hsu et al., 1991 ) Plasmid Description Source pPJ131 E. coli expression vector ( Lu et al., 2006 ) p Ec PlsC E. coli plsC in pPJ131 ( Robertson et al., 2017 ) p Ct PlsC Chlamydia trachomatis plsC in pPJ131 ( Robertson et al., 2017 ) p Af PlsC A. finegoldii plsC ( Alfi_2876 ) in pPJ131 This study p Ec FadD E. coli fadD in pPJ131 ( Yao et al., 2016 ) p Ng Aas Neisseria gonorrhoeae aas in pPJ131 ( Yao et al., 2016 ) p Af Aas1 A. finegoldii aas1 ( Alfi_2635 ) in pPJ131 This study p Af 3181 A. finegoldii Alfi_3181 in pPJ131 This study p Af Aas2 A. finegoldii aas2 ( Alfi_0371 ) in pPJ131 This study pET_ Af Aas1 A. finegoldii aas1 ( Alfi_2635 ) in pET This study pET_ Af Aas2 A. finegoldii aas2 ( Alfi_0371 ) in pET This study Open in a separate window Bacterial strains and plasmids The minimum inhibitory concentration for cerulenin with or without exogenous fatty acid supplementation against A. finegoldii was determined using a modified broth microdilution method ( Parsons et al., 2011 ).

Techniques: Labeling, Mass Spectrometry

Journal: Molecular microbiology

Article Title: Fatty acid activation and utilization by Alistipes finegoldii , a representative Bacteroidetes resident of the human gut microbiome

doi: 10.1111/mmi.14445

Figure Lengend Snippet: A. finegoldii was labeled with either [d3]14:0 or [d4]16:0, and the lipids were extracted as described under “Experimental Procedures.” Mass spectra of the SL were obtained in both cases and fragmented to determine the positional distribution of the incorporated fatty acids into the SL molecule. R1 of SL is the fatty acid condensed with cysteic acid, and R2 is the fatty acid attached to the amine.

Article Snippet: The A 600 in the 16×100 mm borosilicate glass tubes were measured using a Biochrom WPA CO8000 cell density meter. table ft1 table-wrap mode="anchored" t5 caption a7 Strain Relevant Genotype Source A. finegoldii Strain DSM 17242 (DSMZ Collection) ( Rautio et al., 2003 ) E. coli SM2-1 plsC101 (Ts) metCl6S :: Tn10 ( Coleman, 1990 ) E. coli LCH30 aas-1 fadD88 metB1 relA1 spoT1 ( Hsu et al., 1991 ) Plasmid Description Source pPJ131 E. coli expression vector ( Lu et al., 2006 ) p Ec PlsC E. coli plsC in pPJ131 ( Robertson et al., 2017 ) p Ct PlsC Chlamydia trachomatis plsC in pPJ131 ( Robertson et al., 2017 ) p Af PlsC A. finegoldii plsC ( Alfi_2876 ) in pPJ131 This study p Ec FadD E. coli fadD in pPJ131 ( Yao et al., 2016 ) p Ng Aas Neisseria gonorrhoeae aas in pPJ131 ( Yao et al., 2016 ) p Af Aas1 A. finegoldii aas1 ( Alfi_2635 ) in pPJ131 This study p Af 3181 A. finegoldii Alfi_3181 in pPJ131 This study p Af Aas2 A. finegoldii aas2 ( Alfi_0371 ) in pPJ131 This study pET_ Af Aas1 A. finegoldii aas1 ( Alfi_2635 ) in pET This study pET_ Af Aas2 A. finegoldii aas2 ( Alfi_0371 ) in pET This study Open in a separate window Bacterial strains and plasmids The minimum inhibitory concentration for cerulenin with or without exogenous fatty acid supplementation against A. finegoldii was determined using a modified broth microdilution method ( Parsons et al., 2011 ).

Techniques: Labeling

Journal: Molecular microbiology

Article Title: Fatty acid activation and utilization by Alistipes finegoldii , a representative Bacteroidetes resident of the human gut microbiome

doi: 10.1111/mmi.14445

Figure Lengend Snippet: A. Pathway for the synthesis of PE in A. finegoldii. Acyl-ACP is formed from fatty acid synthesis (FASII) or by the activation of an exogenous fatty acid (FA). The acyl-ACP is converted to acyl-PO4 by PlsX, and the 1-position of glycerol-phosphate (G3P) is acylated by PlsY to form 1-acyl-sn-glycero-3-phosphate (LPA). PlsC acylates the 2-position with an acyl-ACP to form phosphatidic acid (PA) that is converted to CDP-dacylglycerol (CDP-DAG) by CdsA. PssA converts PA to phosphatidylserine (PS), which is then decarboxylated by Psd to form PE.

Article Snippet: The A 600 in the 16×100 mm borosilicate glass tubes were measured using a Biochrom WPA CO8000 cell density meter. table ft1 table-wrap mode="anchored" t5 caption a7 Strain Relevant Genotype Source A. finegoldii Strain DSM 17242 (DSMZ Collection) ( Rautio et al., 2003 ) E. coli SM2-1 plsC101 (Ts) metCl6S :: Tn10 ( Coleman, 1990 ) E. coli LCH30 aas-1 fadD88 metB1 relA1 spoT1 ( Hsu et al., 1991 ) Plasmid Description Source pPJ131 E. coli expression vector ( Lu et al., 2006 ) p Ec PlsC E. coli plsC in pPJ131 ( Robertson et al., 2017 ) p Ct PlsC Chlamydia trachomatis plsC in pPJ131 ( Robertson et al., 2017 ) p Af PlsC A. finegoldii plsC ( Alfi_2876 ) in pPJ131 This study p Ec FadD E. coli fadD in pPJ131 ( Yao et al., 2016 ) p Ng Aas Neisseria gonorrhoeae aas in pPJ131 ( Yao et al., 2016 ) p Af Aas1 A. finegoldii aas1 ( Alfi_2635 ) in pPJ131 This study p Af 3181 A. finegoldii Alfi_3181 in pPJ131 This study p Af Aas2 A. finegoldii aas2 ( Alfi_0371 ) in pPJ131 This study pET_ Af Aas1 A. finegoldii aas1 ( Alfi_2635 ) in pET This study pET_ Af Aas2 A. finegoldii aas2 ( Alfi_0371 ) in pET This study Open in a separate window Bacterial strains and plasmids The minimum inhibitory concentration for cerulenin with or without exogenous fatty acid supplementation against A. finegoldii was determined using a modified broth microdilution method ( Parsons et al., 2011 ).

Techniques: Activation Assay

Journal: Molecular microbiology

Article Title: Fatty acid activation and utilization by Alistipes finegoldii , a representative Bacteroidetes resident of the human gut microbiome

doi: 10.1111/mmi.14445

Figure Lengend Snippet: A. finegoldii uses two acyl-ACP synthetases with different substrate specificities to activate a spectrum of exogenous fatty acid chain lengths found in the gut environment. R1 and R2 refer to the 1- and 2-positions of PE, respectively. R1 of SL is the fatty acid condensed with cysteic acid, and R2 is the fatty acid attached to the amine (see Fig. 4C for structure). Medium-chain fatty acids <16 carbons are ligated to ACP by AfAas1. These medium-chain acyl-ACPs are elongated by FASII prior to the incorporation of the elongated acyl-ACP into both the R1 and R2 positions of both PE and SL. Long-chain fatty acids ≥16 carbons are activated by AfAas2. Acyl-ACP 16 carbons or longer cannot enter FASII and are funneled into the R2 position of PE at the PlsC step. AfAas2 can also use medium-chain fatty acids as substrates, and these acyl-ACP are handled in the same manner as medium-chain acyl-ACP derived from AfAas1. The only long-chain acyl-ACP used for SL synthesis is 16:0-ACP, and it is incorporated into both the R1 and R2 positions of SL.

Article Snippet: The A 600 in the 16×100 mm borosilicate glass tubes were measured using a Biochrom WPA CO8000 cell density meter. table ft1 table-wrap mode="anchored" t5 caption a7 Strain Relevant Genotype Source A. finegoldii Strain DSM 17242 (DSMZ Collection) ( Rautio et al., 2003 ) E. coli SM2-1 plsC101 (Ts) metCl6S :: Tn10 ( Coleman, 1990 ) E. coli LCH30 aas-1 fadD88 metB1 relA1 spoT1 ( Hsu et al., 1991 ) Plasmid Description Source pPJ131 E. coli expression vector ( Lu et al., 2006 ) p Ec PlsC E. coli plsC in pPJ131 ( Robertson et al., 2017 ) p Ct PlsC Chlamydia trachomatis plsC in pPJ131 ( Robertson et al., 2017 ) p Af PlsC A. finegoldii plsC ( Alfi_2876 ) in pPJ131 This study p Ec FadD E. coli fadD in pPJ131 ( Yao et al., 2016 ) p Ng Aas Neisseria gonorrhoeae aas in pPJ131 ( Yao et al., 2016 ) p Af Aas1 A. finegoldii aas1 ( Alfi_2635 ) in pPJ131 This study p Af 3181 A. finegoldii Alfi_3181 in pPJ131 This study p Af Aas2 A. finegoldii aas2 ( Alfi_0371 ) in pPJ131 This study pET_ Af Aas1 A. finegoldii aas1 ( Alfi_2635 ) in pET This study pET_ Af Aas2 A. finegoldii aas2 ( Alfi_0371 ) in pET This study Open in a separate window Bacterial strains and plasmids The minimum inhibitory concentration for cerulenin with or without exogenous fatty acid supplementation against A. finegoldii was determined using a modified broth microdilution method ( Parsons et al., 2011 ).

Techniques: Derivative Assay

Journal: Molecular microbiology

Article Title: Fatty acid activation and utilization by Alistipes finegoldii , a representative Bacteroidetes resident of the human gut microbiome

doi: 10.1111/mmi.14445

Figure Lengend Snippet: Bacterial strains and plasmids

Article Snippet: The A 600 in the 16×100 mm borosilicate glass tubes were measured using a Biochrom WPA CO8000 cell density meter. table ft1 table-wrap mode="anchored" t5 caption a7 Strain Relevant Genotype Source A. finegoldii Strain DSM 17242 (DSMZ Collection) ( Rautio et al., 2003 ) E. coli SM2-1 plsC101 (Ts) metCl6S :: Tn10 ( Coleman, 1990 ) E. coli LCH30 aas-1 fadD88 metB1 relA1 spoT1 ( Hsu et al., 1991 ) Plasmid Description Source pPJ131 E. coli expression vector ( Lu et al., 2006 ) p Ec PlsC E. coli plsC in pPJ131 ( Robertson et al., 2017 ) p Ct PlsC Chlamydia trachomatis plsC in pPJ131 ( Robertson et al., 2017 ) p Af PlsC A. finegoldii plsC ( Alfi_2876 ) in pPJ131 This study p Ec FadD E. coli fadD in pPJ131 ( Yao et al., 2016 ) p Ng Aas Neisseria gonorrhoeae aas in pPJ131 ( Yao et al., 2016 ) p Af Aas1 A. finegoldii aas1 ( Alfi_2635 ) in pPJ131 This study p Af 3181 A. finegoldii Alfi_3181 in pPJ131 This study p Af Aas2 A. finegoldii aas2 ( Alfi_0371 ) in pPJ131 This study pET_ Af Aas1 A. finegoldii aas1 ( Alfi_2635 ) in pET This study pET_ Af Aas2 A. finegoldii aas2 ( Alfi_0371 ) in pET This study Open in a separate window Bacterial strains and plasmids The minimum inhibitory concentration for cerulenin with or without exogenous fatty acid supplementation against A. finegoldii was determined using a modified broth microdilution method ( Parsons et al., 2011 ).

Techniques: