Journal: Biotechnology for Biofuels and Bioproducts

Article Title: Isopropanol production via the thermophilic bioconversion of sugars and syngas using metabolically engineered Moorella thermoacetica

doi: 10.1186/s13068-024-02460-1

Figure Lengend Snippet: Introduction of sadh in M. thermoacetica . A Schematic representation of plasmid construction and sadh introduction in place of pduL2 . B Agarose gel electrophoresis following PCR amplification of the pduL2 region of the host and a recombinant strain. M, DNA size marker; 1, the pduL2::sadh strain; 2, the ∆ pyrF strain. The size of the amplified region was shifted to 2.6 kb in the pduL2::sadh strain, consistent with the DNA construct, which was originally 0.9 kb in the ∆ pyrF strain. C A plausible metabolic pathway showing cofactor supply for ethanol production from hexose. Reducing equivalents NADH and reduced Fd were converted into NADPH via enzymatic electron confurcation. NADPH is a cofactor for Sadh. D Culture profile of the pduL2::sadh strain supplemented with fructose as the substrate. E Acetone supplementation to the culture of the pduL2::sadh strain. The condition was the same as in D . Data are presented as the mean with SDs of two biological replicates in D , E . Most error bars are smaller than the symbols of data plots

Article Snippet: The plasmid construct to introduce sadh in place of pduL2 was constructed. sadh DNA sequence was codon-optimized for M. thermoacetica and synthesized (Genewiz).

Techniques: Plasmid Preparation, Agarose Gel Electrophoresis, Amplification, Recombinant, Marker, Construct

Journal: Biotechnology for Biofuels and Bioproducts

Article Title: Isopropanol production via the thermophilic bioconversion of sugars and syngas using metabolically engineered Moorella thermoacetica

doi: 10.1186/s13068-024-02460-1

Figure Lengend Snippet: Introduction of genes for IPA production in M. thermoacetica . A Schematic representation of the synthetic IPA operon encoding enzymes for IPA production from acetyl-CoA. The reactions by enzymes derived from corresponding genes are shown in Fig. . B Schematic representation of the plasmid construction and introduction of the synthetic IPA operon in place of pduL2 . C Agarose gel electrophoresis, following PCR amplification of the pduL2 region of the host and recombinant strains. M, DNA size marker; 1, the ∆ pyrF strain; 2, the plasmid pHM71 (positive control); 3–8, candidates of the pduL2::IPA strain. The size of the amplified region was shifted to 6.5 kb in the pduL2::IPA strain, consistent with the DNA construct, which was originally 0.9 kb in the ∆ pyrF strain. A candidate clone (lane No. 3) showed a faint band with the same migration as the ∆ pyrF strain, which indicated a mixed population. This clone was excluded. The other clones showed the same culture profile in the following fermentation analysis

Article Snippet: The plasmid construct to introduce sadh in place of pduL2 was constructed. sadh DNA sequence was codon-optimized for M. thermoacetica and synthesized (Genewiz).

Techniques: Derivative Assay, Plasmid Preparation, Agarose Gel Electrophoresis, Amplification, Recombinant, Marker, Positive Control, Construct, Migration, Clone Assay

Journal: Biotechnology for Biofuels and Bioproducts

Article Title: Isopropanol production via the thermophilic bioconversion of sugars and syngas using metabolically engineered Moorella thermoacetica

doi: 10.1186/s13068-024-02460-1

Figure Lengend Snippet: Strains and plasmids used in this study

Article Snippet: The plasmid construct to introduce sadh in place of pduL2 was constructed. sadh DNA sequence was codon-optimized for M. thermoacetica and synthesized (Genewiz).

Techniques: Plasmid Preparation, Clone Assay, Modification, Methylation, DNA Methylation Assay, Transformation Assay, Introduce, Construct, Marker