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Membrane potential limits growth and substrate utilization in C. bescii, alleviation dramatically improves productivity

ABSTRACT: Membrane potential limits growth and substrate utilization in C. bescii, alleviation dramatically improves productivity

PROVIDER: PRJNA430890 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6497360.fastq.gz	Fastqsanger.gz
	SRR6497370.fastq.gz	Fastqsanger.gz
	SRR6497380.fastq.gz	Fastqsanger.gz
	SRR6497361.fastq.gz	Fastqsanger.gz
	SRR6497371.fastq.gz	Fastqsanger.gz

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Membrane potential limits growth and substrate utilization in C. bescii, alleviation dramatically improves productivity

Project description:Caldicellulosiruptor bescii is an anaerobic hyper thermophile that can utilize a wide range of substrates. However, inhibitors released from biomass can result in unfavorable growth conditions and limit bioconversion to products. Medium as well as intracellular pH are conditions critical for growth and prone to change in effect of fermentation end or by products such as, CO2, organic acids etc. Growth pH for C. bescii as currently reported is a narrow range of 6.8-7.3. In this study, we examined the physiological and systems level responses of C. bescii to growth at acidic pH. Samples collected from bottles, controlled batch, fed-batch and chemostat systems were subjected to growth, product and integrated omics profiling. It was discovered that in batch reactors, lowering pH from 7.2 to 6.0 at the mid-log phase, led to a significant increase in growth and product yields. Time course transcriptomics data collected from these batch reactors was analyzed to try and get a better understanding of the underlying mechanisms for improved growth.

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Project description:Extremely thermophilic bacteria from the genus Caldicellulosiruptor can degrade various polysaccharide components of plant cell walls. Previous experimental studies identified a variety of carbohydrate-active enzymes in model species C. saccharolyticus and C.bescii, while prior transcriptomic experiments identified their putative carbohydrate uptake transporters. We investigated the mechanisms of transcriptional regulation of carbohydrate utilization genes using a comparative genomics approach applied to fourteen Caldicellulosiruptor species. The reconstruction of carbohydrate utilization regulatory network includes the predicted binding sites for 34 mostly local regulators and point to the regulatory mechanisms controlling expression of genes involved in degradation of plant biomass. The Rex and CggR regulons control the central glycolytic and primary redox reactions. The identified transcription factor binding sites and regulons were validated with transcriptomic and transcription start site data for C. bescii grown on cellulose, cellobiose, glucose, xylan, and xylose. The XylR and XynR regulons control xylan-induced transcriptional response of genes involved in degradation of xylan and xylose utilization. The reconstructed regulons informed the carbohydrate utilization reconstruction analysis and improved functional annotations of 51 transporters and 11 catabolic enzymes. Using gene deletion, we confirmed that the shared ATPase component MsmK is essential for growth on oligo- and polysaccharides but not for the utilization of monosaccharides. By elucidating the carbohydrate utilization framework in C. bescii, strategies for metabolic engineering can be pursued to optimize yields of bio-based fuels and chemicals from lignocellulose.

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