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Production pathway and precursors of NMN in <t>L.</t> <t>reuteri</t> JCM 1112 T . ( A ) Putative NMN production pathway in bacteria ( , ). FtNadE: NMN synthase of Francisella tularensis ; MazG: nucleoside triphosphate pyrophosphohydrolase; NadC: quinolinate phosphoribosyl transferase; NadD: NAMN adenylyl transferase; NadE: NAD⁺ synthetase; <t>NadV:</t> NAM phosphoribosyl transferase (NAMPT); NudE: NADH hydrolase; PncA: nicotinamidase; PncB: nicotinic acid phosphoribosyl transferase; UshA: UDP-sugar hydrolase. Dotted lines from Asp and Trp to quinolinic acid represent multiple reactions. ( B–F ) Production of labeled NMN by L. reuteri JCM 1112 T in MRS broth supplemented with 13 C 6 -NA ( B ), 13 C 6 -NAM ( C ), 13 C 5 -NAD + ( D ), 13 C 4 - 15 N-Asp ( E ), or 13 C - 15 N 2 -Trp ( F ). The cultivation time was 12 h. The blank bars represent the percentages of labeled compounds (with respect to the total of labeled and unlabeled compounds) at the start of cultivation. The gray bars represent the percentages of labeled NMN (with respect to the total of labeled and unlabeled NMN) in the culture supernatants. The percentages of labeled NA, NAM, and NAD + were calculated from the peak areas of each compound. The percentages of labeled Trp and Asp were calculated based on their respective concentrations of added labeled compounds and the unlabeled concentrations originally present in the MRS broth. ( G ) NMN concentrations in the supernatants of MRS broth cultures of L. reuteri JCM 1112 T supplemented with 5,000 ng/mL of NAD + (7.5 µmol/L), NAM (41 µmol/L), or NA (41 µmol/L). The data are expressed as fold changes in NMN production relative to the unsupplemented MRS broth culture (Control). The cultivation time was 12 h. The bars represent the mean values, and the error bars indicate the standard deviation calculated from triplicates. One-way ANOVA showed a significant treatment effect ( F (3,8) = 570, P < 0.0001). Dunnett’s test was then performed using unsupplemented MRS broth culture as the control; *** P < 0.001 versus control; ns = not significant.
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1) Product Images from "Nicotinamide mononucleotide production by non-recombinant Limosilactobacillus reuteri and nicotinamide adenine dinucleotide synthesizing lactic acid bacteria"

Article Title: Nicotinamide mononucleotide production by non-recombinant Limosilactobacillus reuteri and nicotinamide adenine dinucleotide synthesizing lactic acid bacteria

Journal: Microbiology Spectrum

doi: 10.1128/spectrum.00333-25

Production pathway and precursors of NMN in L. reuteri JCM 1112 T . ( A ) Putative NMN production pathway in bacteria ( , ). FtNadE: NMN synthase of Francisella tularensis ; MazG: nucleoside triphosphate pyrophosphohydrolase; NadC: quinolinate phosphoribosyl transferase; NadD: NAMN adenylyl transferase; NadE: NAD⁺ synthetase; NadV: NAM phosphoribosyl transferase (NAMPT); NudE: NADH hydrolase; PncA: nicotinamidase; PncB: nicotinic acid phosphoribosyl transferase; UshA: UDP-sugar hydrolase. Dotted lines from Asp and Trp to quinolinic acid represent multiple reactions. ( B–F ) Production of labeled NMN by L. reuteri JCM 1112 T in MRS broth supplemented with 13 C 6 -NA ( B ), 13 C 6 -NAM ( C ), 13 C 5 -NAD + ( D ), 13 C 4 - 15 N-Asp ( E ), or 13 C - 15 N 2 -Trp ( F ). The cultivation time was 12 h. The blank bars represent the percentages of labeled compounds (with respect to the total of labeled and unlabeled compounds) at the start of cultivation. The gray bars represent the percentages of labeled NMN (with respect to the total of labeled and unlabeled NMN) in the culture supernatants. The percentages of labeled NA, NAM, and NAD + were calculated from the peak areas of each compound. The percentages of labeled Trp and Asp were calculated based on their respective concentrations of added labeled compounds and the unlabeled concentrations originally present in the MRS broth. ( G ) NMN concentrations in the supernatants of MRS broth cultures of L. reuteri JCM 1112 T supplemented with 5,000 ng/mL of NAD + (7.5 µmol/L), NAM (41 µmol/L), or NA (41 µmol/L). The data are expressed as fold changes in NMN production relative to the unsupplemented MRS broth culture (Control). The cultivation time was 12 h. The bars represent the mean values, and the error bars indicate the standard deviation calculated from triplicates. One-way ANOVA showed a significant treatment effect ( F (3,8) = 570, P < 0.0001). Dunnett’s test was then performed using unsupplemented MRS broth culture as the control; *** P < 0.001 versus control; ns = not significant.
Figure Legend Snippet: Production pathway and precursors of NMN in L. reuteri JCM 1112 T . ( A ) Putative NMN production pathway in bacteria ( , ). FtNadE: NMN synthase of Francisella tularensis ; MazG: nucleoside triphosphate pyrophosphohydrolase; NadC: quinolinate phosphoribosyl transferase; NadD: NAMN adenylyl transferase; NadE: NAD⁺ synthetase; NadV: NAM phosphoribosyl transferase (NAMPT); NudE: NADH hydrolase; PncA: nicotinamidase; PncB: nicotinic acid phosphoribosyl transferase; UshA: UDP-sugar hydrolase. Dotted lines from Asp and Trp to quinolinic acid represent multiple reactions. ( B–F ) Production of labeled NMN by L. reuteri JCM 1112 T in MRS broth supplemented with 13 C 6 -NA ( B ), 13 C 6 -NAM ( C ), 13 C 5 -NAD + ( D ), 13 C 4 - 15 N-Asp ( E ), or 13 C - 15 N 2 -Trp ( F ). The cultivation time was 12 h. The blank bars represent the percentages of labeled compounds (with respect to the total of labeled and unlabeled compounds) at the start of cultivation. The gray bars represent the percentages of labeled NMN (with respect to the total of labeled and unlabeled NMN) in the culture supernatants. The percentages of labeled NA, NAM, and NAD + were calculated from the peak areas of each compound. The percentages of labeled Trp and Asp were calculated based on their respective concentrations of added labeled compounds and the unlabeled concentrations originally present in the MRS broth. ( G ) NMN concentrations in the supernatants of MRS broth cultures of L. reuteri JCM 1112 T supplemented with 5,000 ng/mL of NAD + (7.5 µmol/L), NAM (41 µmol/L), or NA (41 µmol/L). The data are expressed as fold changes in NMN production relative to the unsupplemented MRS broth culture (Control). The cultivation time was 12 h. The bars represent the mean values, and the error bars indicate the standard deviation calculated from triplicates. One-way ANOVA showed a significant treatment effect ( F (3,8) = 570, P < 0.0001). Dunnett’s test was then performed using unsupplemented MRS broth culture as the control; *** P < 0.001 versus control; ns = not significant.

Techniques Used: Bacteria, Labeling, Control, Standard Deviation



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Production pathway and precursors of NMN in <t>L.</t> <t>reuteri</t> JCM 1112 T . ( A ) Putative NMN production pathway in bacteria ( , ). FtNadE: NMN synthase of Francisella tularensis ; MazG: nucleoside triphosphate pyrophosphohydrolase; NadC: quinolinate phosphoribosyl transferase; NadD: NAMN adenylyl transferase; NadE: NAD⁺ synthetase; <t>NadV:</t> NAM phosphoribosyl transferase (NAMPT); NudE: NADH hydrolase; PncA: nicotinamidase; PncB: nicotinic acid phosphoribosyl transferase; UshA: UDP-sugar hydrolase. Dotted lines from Asp and Trp to quinolinic acid represent multiple reactions. ( B–F ) Production of labeled NMN by L. reuteri JCM 1112 T in MRS broth supplemented with 13 C 6 -NA ( B ), 13 C 6 -NAM ( C ), 13 C 5 -NAD + ( D ), 13 C 4 - 15 N-Asp ( E ), or 13 C - 15 N 2 -Trp ( F ). The cultivation time was 12 h. The blank bars represent the percentages of labeled compounds (with respect to the total of labeled and unlabeled compounds) at the start of cultivation. The gray bars represent the percentages of labeled NMN (with respect to the total of labeled and unlabeled NMN) in the culture supernatants. The percentages of labeled NA, NAM, and NAD + were calculated from the peak areas of each compound. The percentages of labeled Trp and Asp were calculated based on their respective concentrations of added labeled compounds and the unlabeled concentrations originally present in the MRS broth. ( G ) NMN concentrations in the supernatants of MRS broth cultures of L. reuteri JCM 1112 T supplemented with 5,000 ng/mL of NAD + (7.5 µmol/L), NAM (41 µmol/L), or NA (41 µmol/L). The data are expressed as fold changes in NMN production relative to the unsupplemented MRS broth culture (Control). The cultivation time was 12 h. The bars represent the mean values, and the error bars indicate the standard deviation calculated from triplicates. One-way ANOVA showed a significant treatment effect ( F (3,8) = 570, P < 0.0001). Dunnett’s test was then performed using unsupplemented MRS broth culture as the control; *** P < 0.001 versus control; ns = not significant.
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Production pathway and precursors of NMN in <t>L.</t> <t>reuteri</t> JCM 1112 T . ( A ) Putative NMN production pathway in bacteria ( , ). FtNadE: NMN synthase of Francisella tularensis ; MazG: nucleoside triphosphate pyrophosphohydrolase; NadC: quinolinate phosphoribosyl transferase; NadD: NAMN adenylyl transferase; NadE: NAD⁺ synthetase; <t>NadV:</t> NAM phosphoribosyl transferase (NAMPT); NudE: NADH hydrolase; PncA: nicotinamidase; PncB: nicotinic acid phosphoribosyl transferase; UshA: UDP-sugar hydrolase. Dotted lines from Asp and Trp to quinolinic acid represent multiple reactions. ( B–F ) Production of labeled NMN by L. reuteri JCM 1112 T in MRS broth supplemented with 13 C 6 -NA ( B ), 13 C 6 -NAM ( C ), 13 C 5 -NAD + ( D ), 13 C 4 - 15 N-Asp ( E ), or 13 C - 15 N 2 -Trp ( F ). The cultivation time was 12 h. The blank bars represent the percentages of labeled compounds (with respect to the total of labeled and unlabeled compounds) at the start of cultivation. The gray bars represent the percentages of labeled NMN (with respect to the total of labeled and unlabeled NMN) in the culture supernatants. The percentages of labeled NA, NAM, and NAD + were calculated from the peak areas of each compound. The percentages of labeled Trp and Asp were calculated based on their respective concentrations of added labeled compounds and the unlabeled concentrations originally present in the MRS broth. ( G ) NMN concentrations in the supernatants of MRS broth cultures of L. reuteri JCM 1112 T supplemented with 5,000 ng/mL of NAD + (7.5 µmol/L), NAM (41 µmol/L), or NA (41 µmol/L). The data are expressed as fold changes in NMN production relative to the unsupplemented MRS broth culture (Control). The cultivation time was 12 h. The bars represent the mean values, and the error bars indicate the standard deviation calculated from triplicates. One-way ANOVA showed a significant treatment effect ( F (3,8) = 570, P < 0.0001). Dunnett’s test was then performed using unsupplemented MRS broth culture as the control; *** P < 0.001 versus control; ns = not significant.
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Production pathway and precursors of NMN in <t>L.</t> <t>reuteri</t> JCM 1112 T . ( A ) Putative NMN production pathway in bacteria ( , ). FtNadE: NMN synthase of Francisella tularensis ; MazG: nucleoside triphosphate pyrophosphohydrolase; NadC: quinolinate phosphoribosyl transferase; NadD: NAMN adenylyl transferase; NadE: NAD⁺ synthetase; <t>NadV:</t> NAM phosphoribosyl transferase (NAMPT); NudE: NADH hydrolase; PncA: nicotinamidase; PncB: nicotinic acid phosphoribosyl transferase; UshA: UDP-sugar hydrolase. Dotted lines from Asp and Trp to quinolinic acid represent multiple reactions. ( B–F ) Production of labeled NMN by L. reuteri JCM 1112 T in MRS broth supplemented with 13 C 6 -NA ( B ), 13 C 6 -NAM ( C ), 13 C 5 -NAD + ( D ), 13 C 4 - 15 N-Asp ( E ), or 13 C - 15 N 2 -Trp ( F ). The cultivation time was 12 h. The blank bars represent the percentages of labeled compounds (with respect to the total of labeled and unlabeled compounds) at the start of cultivation. The gray bars represent the percentages of labeled NMN (with respect to the total of labeled and unlabeled NMN) in the culture supernatants. The percentages of labeled NA, NAM, and NAD + were calculated from the peak areas of each compound. The percentages of labeled Trp and Asp were calculated based on their respective concentrations of added labeled compounds and the unlabeled concentrations originally present in the MRS broth. ( G ) NMN concentrations in the supernatants of MRS broth cultures of L. reuteri JCM 1112 T supplemented with 5,000 ng/mL of NAD + (7.5 µmol/L), NAM (41 µmol/L), or NA (41 µmol/L). The data are expressed as fold changes in NMN production relative to the unsupplemented MRS broth culture (Control). The cultivation time was 12 h. The bars represent the mean values, and the error bars indicate the standard deviation calculated from triplicates. One-way ANOVA showed a significant treatment effect ( F (3,8) = 570, P < 0.0001). Dunnett’s test was then performed using unsupplemented MRS broth culture as the control; *** P < 0.001 versus control; ns = not significant.
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Production pathway and precursors of NMN in <t>L.</t> <t>reuteri</t> JCM 1112 T . ( A ) Putative NMN production pathway in bacteria ( , ). FtNadE: NMN synthase of Francisella tularensis ; MazG: nucleoside triphosphate pyrophosphohydrolase; NadC: quinolinate phosphoribosyl transferase; NadD: NAMN adenylyl transferase; NadE: NAD⁺ synthetase; <t>NadV:</t> NAM phosphoribosyl transferase (NAMPT); NudE: NADH hydrolase; PncA: nicotinamidase; PncB: nicotinic acid phosphoribosyl transferase; UshA: UDP-sugar hydrolase. Dotted lines from Asp and Trp to quinolinic acid represent multiple reactions. ( B–F ) Production of labeled NMN by L. reuteri JCM 1112 T in MRS broth supplemented with 13 C 6 -NA ( B ), 13 C 6 -NAM ( C ), 13 C 5 -NAD + ( D ), 13 C 4 - 15 N-Asp ( E ), or 13 C - 15 N 2 -Trp ( F ). The cultivation time was 12 h. The blank bars represent the percentages of labeled compounds (with respect to the total of labeled and unlabeled compounds) at the start of cultivation. The gray bars represent the percentages of labeled NMN (with respect to the total of labeled and unlabeled NMN) in the culture supernatants. The percentages of labeled NA, NAM, and NAD + were calculated from the peak areas of each compound. The percentages of labeled Trp and Asp were calculated based on their respective concentrations of added labeled compounds and the unlabeled concentrations originally present in the MRS broth. ( G ) NMN concentrations in the supernatants of MRS broth cultures of L. reuteri JCM 1112 T supplemented with 5,000 ng/mL of NAD + (7.5 µmol/L), NAM (41 µmol/L), or NA (41 µmol/L). The data are expressed as fold changes in NMN production relative to the unsupplemented MRS broth culture (Control). The cultivation time was 12 h. The bars represent the mean values, and the error bars indicate the standard deviation calculated from triplicates. One-way ANOVA showed a significant treatment effect ( F (3,8) = 570, P < 0.0001). Dunnett’s test was then performed using unsupplemented MRS broth culture as the control; *** P < 0.001 versus control; ns = not significant.
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Production pathway and precursors of NMN in L. reuteri JCM 1112 T . ( A ) Putative NMN production pathway in bacteria ( , ). FtNadE: NMN synthase of Francisella tularensis ; MazG: nucleoside triphosphate pyrophosphohydrolase; NadC: quinolinate phosphoribosyl transferase; NadD: NAMN adenylyl transferase; NadE: NAD⁺ synthetase; NadV: NAM phosphoribosyl transferase (NAMPT); NudE: NADH hydrolase; PncA: nicotinamidase; PncB: nicotinic acid phosphoribosyl transferase; UshA: UDP-sugar hydrolase. Dotted lines from Asp and Trp to quinolinic acid represent multiple reactions. ( B–F ) Production of labeled NMN by L. reuteri JCM 1112 T in MRS broth supplemented with 13 C 6 -NA ( B ), 13 C 6 -NAM ( C ), 13 C 5 -NAD + ( D ), 13 C 4 - 15 N-Asp ( E ), or 13 C - 15 N 2 -Trp ( F ). The cultivation time was 12 h. The blank bars represent the percentages of labeled compounds (with respect to the total of labeled and unlabeled compounds) at the start of cultivation. The gray bars represent the percentages of labeled NMN (with respect to the total of labeled and unlabeled NMN) in the culture supernatants. The percentages of labeled NA, NAM, and NAD + were calculated from the peak areas of each compound. The percentages of labeled Trp and Asp were calculated based on their respective concentrations of added labeled compounds and the unlabeled concentrations originally present in the MRS broth. ( G ) NMN concentrations in the supernatants of MRS broth cultures of L. reuteri JCM 1112 T supplemented with 5,000 ng/mL of NAD + (7.5 µmol/L), NAM (41 µmol/L), or NA (41 µmol/L). The data are expressed as fold changes in NMN production relative to the unsupplemented MRS broth culture (Control). The cultivation time was 12 h. The bars represent the mean values, and the error bars indicate the standard deviation calculated from triplicates. One-way ANOVA showed a significant treatment effect ( F (3,8) = 570, P < 0.0001). Dunnett’s test was then performed using unsupplemented MRS broth culture as the control; *** P < 0.001 versus control; ns = not significant.

Journal: Microbiology Spectrum

Article Title: Nicotinamide mononucleotide production by non-recombinant Limosilactobacillus reuteri and nicotinamide adenine dinucleotide synthesizing lactic acid bacteria

doi: 10.1128/spectrum.00333-25

Figure Lengend Snippet: Production pathway and precursors of NMN in L. reuteri JCM 1112 T . ( A ) Putative NMN production pathway in bacteria ( , ). FtNadE: NMN synthase of Francisella tularensis ; MazG: nucleoside triphosphate pyrophosphohydrolase; NadC: quinolinate phosphoribosyl transferase; NadD: NAMN adenylyl transferase; NadE: NAD⁺ synthetase; NadV: NAM phosphoribosyl transferase (NAMPT); NudE: NADH hydrolase; PncA: nicotinamidase; PncB: nicotinic acid phosphoribosyl transferase; UshA: UDP-sugar hydrolase. Dotted lines from Asp and Trp to quinolinic acid represent multiple reactions. ( B–F ) Production of labeled NMN by L. reuteri JCM 1112 T in MRS broth supplemented with 13 C 6 -NA ( B ), 13 C 6 -NAM ( C ), 13 C 5 -NAD + ( D ), 13 C 4 - 15 N-Asp ( E ), or 13 C - 15 N 2 -Trp ( F ). The cultivation time was 12 h. The blank bars represent the percentages of labeled compounds (with respect to the total of labeled and unlabeled compounds) at the start of cultivation. The gray bars represent the percentages of labeled NMN (with respect to the total of labeled and unlabeled NMN) in the culture supernatants. The percentages of labeled NA, NAM, and NAD + were calculated from the peak areas of each compound. The percentages of labeled Trp and Asp were calculated based on their respective concentrations of added labeled compounds and the unlabeled concentrations originally present in the MRS broth. ( G ) NMN concentrations in the supernatants of MRS broth cultures of L. reuteri JCM 1112 T supplemented with 5,000 ng/mL of NAD + (7.5 µmol/L), NAM (41 µmol/L), or NA (41 µmol/L). The data are expressed as fold changes in NMN production relative to the unsupplemented MRS broth culture (Control). The cultivation time was 12 h. The bars represent the mean values, and the error bars indicate the standard deviation calculated from triplicates. One-way ANOVA showed a significant treatment effect ( F (3,8) = 570, P < 0.0001). Dunnett’s test was then performed using unsupplemented MRS broth culture as the control; *** P < 0.001 versus control; ns = not significant.

Article Snippet: We could not identify the nadV gene in the L. reuteri genome by a search using L. reuteri and nadV as keywords in the database of the National Center for Biotechnology Information (NCBI Gene, https://www.ncbi.nlm.nih.gov/gene , accessed November 2024).

Techniques: Bacteria, Labeling, Control, Standard Deviation