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Analysis of 23BD production in engineered strains of C. thermocellum . All strains were cultured in MTC-5 medium containing 5 g/L cellobiose as carbon source. (A) 23BD titers detected in the fermentation broth are the sum of both SS/RR 23BD and meso-23BD. (B) ALDC enzyme activity. (C) BDH enzyme activity. For both ALDC and BDH, assay conditions are described in the materials and methods section. Error bars represent the range of data, n = 2 biological replicates. Individual replicate data, and separate quantification of 23BD isomers, is available in Supporting Dataset D2. ∗AG8235 is the parent strain used to construct all the other strains listed in this table Δhpt Δ0478 Δ2366::polyattB. #1 – p1194, promoter from the Clo1313_1194 gene #2 – aggggga ribosome binding site <t>sequence</t> #3 – aggagga ribosome binding site sequence #4 – P_Clo1313_1194 (1st gene in the operon).
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Analysis of 23BD production in engineered strains of C. thermocellum . All strains were cultured in MTC-5 medium containing 5 g/L cellobiose as carbon source. (A) 23BD titers detected in the fermentation broth are the sum of both SS/RR 23BD and meso-23BD. (B) ALDC enzyme activity. (C) BDH enzyme activity. For both ALDC and BDH, assay conditions are described in the materials and methods section. Error bars represent the range of data, n = 2 biological replicates. Individual replicate data, and separate quantification of 23BD isomers, is available in Supporting Dataset D2. ∗AG8235 is the parent strain used to construct all the other strains listed in this table Δhpt Δ0478 Δ2366::polyattB. #1 – p1194, promoter from the Clo1313_1194 gene #2 – aggggga ribosome binding site sequence #3 – aggagga ribosome binding site sequence #4 – P_Clo1313_1194 (1st gene in the operon).

Journal: Metabolic Engineering Communications

Article Title: Engineering Clostridium thermocellum for production of 2,3-butanediol from cellulose ☆

doi: 10.1016/j.mec.2025.e00269

Figure Lengend Snippet: Analysis of 23BD production in engineered strains of C. thermocellum . All strains were cultured in MTC-5 medium containing 5 g/L cellobiose as carbon source. (A) 23BD titers detected in the fermentation broth are the sum of both SS/RR 23BD and meso-23BD. (B) ALDC enzyme activity. (C) BDH enzyme activity. For both ALDC and BDH, assay conditions are described in the materials and methods section. Error bars represent the range of data, n = 2 biological replicates. Individual replicate data, and separate quantification of 23BD isomers, is available in Supporting Dataset D2. ∗AG8235 is the parent strain used to construct all the other strains listed in this table Δhpt Δ0478 Δ2366::polyattB. #1 – p1194, promoter from the Clo1313_1194 gene #2 – aggggga ribosome binding site sequence #3 – aggagga ribosome binding site sequence #4 – P_Clo1313_1194 (1st gene in the operon).

Article Snippet: They were checked for plasmid integration by PCR and whole genome sequencing by Plasmidsaurus Inc.

Techniques: Cell Culture, Activity Assay, Construct, Binding Assay, Sequencing