r ornithinolytica atcc 31898  (ATCC)

Journal: Applied and Environmental Microbiology

Article Title: Metabolic Engineering of Raoultella ornithinolytica BF60 for Production of 2,5-Furandicarboxylic Acid from 5-Hydroxymethylfurfural

doi: 10.1128/AEM.02312-16

Figure Lengend Snippet: Activity of FDCA decarboxylase (gray bar) and 5-HMF reductase (white bar) in the wild-type strain, the dicarboxylic acid decarboxylase mutant (RTBF60-1), and the dicarboxylic acid decarboxylase and aldehyde reductase mutant (RTBF60-2). The data shown represent the averages ± SD of results of three independent replicates.

Article Snippet: TABLE 2 Strain or plasmid Characteristic(s) a Source or reference Strains Escherichia coli JM109 recA1 endA1 gyrA96 thi hsdR17 supE44 relA1 Δ( lac-proAB )/F′ ( traD36 proAB + lacI q lacZ ΔM15) TaKaRa, Otsu, Japan Escherichia coli BL21(DE3) fhuA2 [ lon ] ompT gal (λ DE3 ) [ dcm] ΔhsdS λ DE3 = λ sBamHIo ΔEcoRI-B int ::( lacI::PlacUV5 :: T7 gene 1) i21 Δnin5 Tiangen, China R. ornithinolytica BF60 Wild type This work RTBF60-1 R. ornithinolytica BF60 derivate, insertional mutant in the dcaD gene encoding dicarboxylic acid decarboxylase This work RTBF60-2 RTBF60-1 derivate, insertional mutant in the aldR gene encoding aldehyde reductase This work RTBF60-3 RTBF60-2 derivate overexpressing the gene encoding aldehyde dehydrogenase 1 of R. ornithinolytica BF60 This work RTBF60-4 RTBF60-2 derivate overexpressing the gene encoding aldehyde dehydrogenase 2 of R. ornithinolytica BF60 This work RTBF60-5 RTBF60-2 derivate overexpressing the gene encoding aldehyde dehydrogenase 3 of R. ornithinolytica BF60 This work R. ornithinolytica ATCC 31898 Wild type ATCC Plasmids pACD4K-C-loxP Cm r Sigma-Aldrich, St. Louis, MO, USA pAR1219 Amp r Sigma-Aldrich, St. Louis, MO, USA pMD19T (Simple) Amp r TaKaRa, Otsu, Japan pTSC-repli pTSC plasmid containing broad-host-range replicon of pBBR1 44 pBBR1MCS2 Kan r 61 Open in a separate window a Amp r , ampicillin resistance; Ch r , chloramphenicol resistance; Kan r , kanamycin resistance.

Techniques: Activity Assay, Mutagenesis

Journal: Applied and Environmental Microbiology

Article Title: Metabolic Engineering of Raoultella ornithinolytica BF60 for Production of 2,5-Furandicarboxylic Acid from 5-Hydroxymethylfurfural

doi: 10.1128/AEM.02312-16

Figure Lengend Snippet: Effect of mutagenesis on the molar conversion efficiency and time profile for product formation. (A) Efficiency of conversion of different mutant and overexpressed strains from 5-HMF to FDCA production (mol/mol). (B) Time profile for the production of FDCA from 100 mM 5-HMF by different whole-cell biocatalysts. Wild type, ○; RTBF60-1, ■; RTBF60-2, Δ; RTBF60-3, •. Data were analyzed using the Student t test. P values of less than 0.05 were considered statistically significant (the P value for comparisons of the wild type to the RTBF60-1 strain was 0.0004; the P value for comparisons of the wild type to the RTBF60-2 strain was 0.0002; and the P value for comparisons of the wild type to the RTBF60-3 strain was 0.0004). The data shown represent the averages ± SD of results of three independent replicates.

Article Snippet: TABLE 2 Strain or plasmid Characteristic(s) a Source or reference Strains Escherichia coli JM109 recA1 endA1 gyrA96 thi hsdR17 supE44 relA1 Δ( lac-proAB )/F′ ( traD36 proAB + lacI q lacZ ΔM15) TaKaRa, Otsu, Japan Escherichia coli BL21(DE3) fhuA2 [ lon ] ompT gal (λ DE3 ) [ dcm] ΔhsdS λ DE3 = λ sBamHIo ΔEcoRI-B int ::( lacI::PlacUV5 :: T7 gene 1) i21 Δnin5 Tiangen, China R. ornithinolytica BF60 Wild type This work RTBF60-1 R. ornithinolytica BF60 derivate, insertional mutant in the dcaD gene encoding dicarboxylic acid decarboxylase This work RTBF60-2 RTBF60-1 derivate, insertional mutant in the aldR gene encoding aldehyde reductase This work RTBF60-3 RTBF60-2 derivate overexpressing the gene encoding aldehyde dehydrogenase 1 of R. ornithinolytica BF60 This work RTBF60-4 RTBF60-2 derivate overexpressing the gene encoding aldehyde dehydrogenase 2 of R. ornithinolytica BF60 This work RTBF60-5 RTBF60-2 derivate overexpressing the gene encoding aldehyde dehydrogenase 3 of R. ornithinolytica BF60 This work R. ornithinolytica ATCC 31898 Wild type ATCC Plasmids pACD4K-C-loxP Cm r Sigma-Aldrich, St. Louis, MO, USA pAR1219 Amp r Sigma-Aldrich, St. Louis, MO, USA pMD19T (Simple) Amp r TaKaRa, Otsu, Japan pTSC-repli pTSC plasmid containing broad-host-range replicon of pBBR1 44 pBBR1MCS2 Kan r 61 Open in a separate window a Amp r , ampicillin resistance; Ch r , chloramphenicol resistance; Kan r , kanamycin resistance.

Techniques: Mutagenesis

Journal: Applied and Environmental Microbiology

Article Title: Metabolic Engineering of Raoultella ornithinolytica BF60 for Production of 2,5-Furandicarboxylic Acid from 5-Hydroxymethylfurfural

doi: 10.1128/AEM.02312-16

Figure Lengend Snippet: Strains and plasmids used in this study

Article Snippet: TABLE 2 Strain or plasmid Characteristic(s) a Source or reference Strains Escherichia coli JM109 recA1 endA1 gyrA96 thi hsdR17 supE44 relA1 Δ( lac-proAB )/F′ ( traD36 proAB + lacI q lacZ ΔM15) TaKaRa, Otsu, Japan Escherichia coli BL21(DE3) fhuA2 [ lon ] ompT gal (λ DE3 ) [ dcm] ΔhsdS λ DE3 = λ sBamHIo ΔEcoRI-B int ::( lacI::PlacUV5 :: T7 gene 1) i21 Δnin5 Tiangen, China R. ornithinolytica BF60 Wild type This work RTBF60-1 R. ornithinolytica BF60 derivate, insertional mutant in the dcaD gene encoding dicarboxylic acid decarboxylase This work RTBF60-2 RTBF60-1 derivate, insertional mutant in the aldR gene encoding aldehyde reductase This work RTBF60-3 RTBF60-2 derivate overexpressing the gene encoding aldehyde dehydrogenase 1 of R. ornithinolytica BF60 This work RTBF60-4 RTBF60-2 derivate overexpressing the gene encoding aldehyde dehydrogenase 2 of R. ornithinolytica BF60 This work RTBF60-5 RTBF60-2 derivate overexpressing the gene encoding aldehyde dehydrogenase 3 of R. ornithinolytica BF60 This work R. ornithinolytica ATCC 31898 Wild type ATCC Plasmids pACD4K-C-loxP Cm r Sigma-Aldrich, St. Louis, MO, USA pAR1219 Amp r Sigma-Aldrich, St. Louis, MO, USA pMD19T (Simple) Amp r TaKaRa, Otsu, Japan pTSC-repli pTSC plasmid containing broad-host-range replicon of pBBR1 44 pBBR1MCS2 Kan r 61 Open in a separate window a Amp r , ampicillin resistance; Ch r , chloramphenicol resistance; Kan r , kanamycin resistance.

Techniques: Plasmid Preparation, Mutagenesis