Journal: bioRxiv

Article Title: Expanding the scope of redox-balance growth coupling techniques with a carbon cofeeding strategy

doi: 10.64898/2026.04.01.713023

Figure Lengend Snippet: Schematic depicting the genetic modifications to enable the overaccumulation of NADPH (Δ pgi , Δ edd , Δ qor , Δ sthA ) and the deletion of aceA to prohibit growth on acetate as a carbon source. Pathways enabling growth on a mixture of acetate and glucose are shown below and highlighted (red: acetaldehyde (strain APEQS_PduP), yellow: 3-HB (strain APEQS_3-HB), blue: mevalonate (strain APEQS_MEV_sa)). b, Simplified metabolic stoichiometries showing only bioavailable carbon and relevant reducing equivalents and assuming all carbon flows to acetyl-CoA (our pathways’ precursor molecule). Simplified stoichiometry of E. coli fermentative metabolism and APEQS metabolism when grown on glucose, showing redox-balanced fermentative or rescue pathways below. Ethanol fermentation requires less acetyl-CoA (green) than is produced from one glucose when redox balanced, reflecting its suitability as a fermentation pathway. Partially reducing pathways consume more acetyl-CoA (red) than is made available per unit glucose when redox balanced, indicating their inability to resolve redox balance in APEQS without acetate co-feeding. c, Unsuccessful growth coupling of strain APEQS_PduP in the absence of acetate (n=3). d, Unsuccessful growth coupling of strain APEQS_3-HB in the absence of acetate (n=3). e, Unsuccessful growth coupling of strain APEQS_MEV_sa in the absence of acetate (n=3). f, Successful growth coupling of strain APEQS_PduP when the strain is grown with additional 100 mM sodium acetate to satisfy stoichiometric constraints (n=3). g, Successful growth coupling of strain APEQS_3-HB when the strain is grown with additional 100 mM sodium acetate to satisfy stoichiometric constraints (n=3). h, Successful growth coupling of strain APEQS_MEV_sa when the strain is grown with additional 100 mM sodium acetate to satisfy stoichiometric constraints (n=3). All experiments were conducted in a MOPS medium containing 2% glucose with or without 100 mM acetate. Various concentrations of IPTG were added to modulate the induction of the three partially reducing pathways (high [IPTG]: blue (0.5 mM for p15A-based A5c backbone; 0.05 mM for ColE1-based pQE backbone), medium [IPTG]: purple (0.05 mM for p15A-based A5c backbone; 0.005 mM for ColE1-based pQE backbone), no IPTG: red). An empty vector control was included to demonstrate growth without leaky expression (orange). All growth experiments were repeated a minimum of 3 times and showed identical results.

Article Snippet: For growth coupling experiments and strain evolution, bacterial strains were grown in a minimal MOPS medium (Teknova M2106) containing 2% glucose (w/v) supplemented with the appropriate antibiotic based on the plasmid(s) present in the strain.

Techniques: Produced, Plasmid Preparation, Control, Expressing

Journal: Molecular Microbiology

Article Title: Late Stage Mannan Metabolism in Cellvibrio japonicus Requires the Combined Action of a Mannosyl‐Glucose Phosphorylase and a Mannobiose Epimerase

doi: 10.1111/mmi.70043

Figure Lengend Snippet: Growth analyses of C. japonicus strains grown in MOPS defined media supplemented with 0.5% (w/v) of (A) mannobiose, (B) mannotriose, (C) mannosyl‐glucose, (D) cellobiose, (E) glucomannan, (F) carob galactomannan, (G) guar galactomannan, or (H) linear mannan. Notably, panels A—D were grown on a separate microplate from panels E—F and are shown together for ease of visualization. Growth analyses were conducted using an EPOCH2 microplate reader (Biotek) in biological triplicate for 48 h. Error bars depict standard deviation but are too small to be observed in some graphs. Complete growth statistics are available in Table .

Article Snippet: C. japonicus strains were cultured in MOPS minimal medium from TekNova (cat. no. M2106), supplemented with 1.32 mM phosphate and 0.2% (w/v) glucose.

Techniques: Standard Deviation

Journal: Molecular Microbiology

Article Title: Late Stage Mannan Metabolism in Cellvibrio japonicus Requires the Combined Action of a Mannosyl‐Glucose Phosphorylase and a Mannobiose Epimerase

doi: 10.1111/mmi.70043

Figure Lengend Snippet: Growth analyses of C. japonicus single gene deletions, intermediate strains, and double ectopic complementation strains grown in MOPS defined media supplemented with 0.5% (w/v) of either (A) mannobiose, (B) mannotriose, (C) glucomannan, (D) carob galactomannan, (E) guar galactomannan, or (F) linear mannan. Panels A, B, and F were grown on a separate microplate from Panels C, D, and E, and are shown here together for ease of visualization. Growth analyses were conducted using an EPOCH2 microplate reader (Biotek) in biological triplicate for 48 h. Error bars depict standard deviation but are too small to be observed in many of the graphs. Complete growth statistics are available in Table .

Article Snippet: C. japonicus strains were cultured in MOPS minimal medium from TekNova (cat. no. M2106), supplemented with 1.32 mM phosphate and 0.2% (w/v) glucose.

Techniques: Standard Deviation

Journal: Molecular Microbiology

Article Title: Late Stage Mannan Metabolism in Cellvibrio japonicus Requires the Combined Action of a Mannosyl‐Glucose Phosphorylase and a Mannobiose Epimerase

doi: 10.1111/mmi.70043

Figure Lengend Snippet: Growth analyses of C. japonicus pJKN5 complement strains grown in MOPS defined media supplemented with 0.5% (w/v) of either (A) mannobiose, (B) mannotriose, (C) glucomannan, (D) carob galactomannan, (E) guar galactomannan, or (F) linear mannan. Notably, panels A, B, and F were grown on a separate microplate from panels C, D, and E and are shown together for ease of visualization. Growth analyses were conducted in an EPOCH2 microplate reader (Biotek) in biological triplicate for 48 h. Error bars depict standard deviation and complete growth statistics are available in Table .

Article Snippet: C. japonicus strains were cultured in MOPS minimal medium from TekNova (cat. no. M2106), supplemented with 1.32 mM phosphate and 0.2% (w/v) glucose.

Techniques: Standard Deviation