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There are two pathways for cellobiose utilization in E. coli.
Further, manual curation was done using the published biochemical literature specifically for cellobiose utilization as explained in Figure 10.
Putative gene clusters for cellobiose utilization were identified by keyword screening of gene annotations for "β-glucosidase", and by BLAST homology searches of each genome using β-glucosidase protein sequences from the Carbohydrate-Active enZYmes database (http://www.cazy.org).org
Combining cellobiose with a probiotic strain of Lactobacillus rhamnosus was previously reported to give a synergistic increase of this and other lactic acid bacteria in the cecum of rats [ 33], but no proteins important for cellobiose utilization by lactic acid bacteria were identified in that study.
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During the first 2 h of cellobiose utilization, a temporary growth cessation was observed.
2-deoxyglucose delayed growth in cellobiose but not glucose cultures, indicating that 2-deoxyglucose led to CCR of cellobiose utilization.
This efficiency of cellobiose utilization is as good as its glucose utilization, When 2% beta-glycan was the sole carbon source, KR5 produced 5.4 g/L ethanol with a 74% conversion ratio in 168 h at 37°C.
By using transcription profiling to identify TFs that are significantly differentially expressed in the two fermentation conditions, we were able to rapidly screen a small number of targets for improved cellobiose utilization.
Engineered E. coli Suc260 was further reconstructed by overexpressing the β-glucosidase for achieving direct cellobiose utilization, which could efficiently utilize the pretreated sugarcane bagasses hydrolysate (SBH) consisting of 25.30 g·L−1 cellobiose, 9.70 glucoselucose, 5.90 g·L−1 arabinose and 7.10 g·L−1 xylose.
Nonetheless, evidence for genome decay in the evolution of the cheese specialist strains UW1 and UW4 was observed in regard to genes for inulin, sucrose and cellobiose utilization.
For this study, S. cerevisiae was engineered to achieve cellobiose utilization by introducing both a cellodextrin transporter gene (cdt-1) and an intracellular β-glucosidase gene (gh1-1) from Neurospora crassa (See Methods).
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