Integrating the extracellular, intracellular, and intercellular metabolic processes of Escherichia coli through glucose saturation, inhibition of the acetyl-CoA carboxylase subunit accA with asRNA, and through quantifying cell to cell quorum-sensing
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Abstract
The study aims to demonstrate the link between bacterial cell metabolism and virulence through integrating the environmental, genetic, and cell to cell signaling molecular processes. Dietary fiber metabolized into glucose, increases the proliferation of intestinal microflora, which augments the outputof the Short Chain Fatty Acids. Bacteria ferment the glucose, from fiber, into Short Chain Fatty Acids, which help regulate many biochemical processes and pathways. Each SCFA maintains colonic pH, promotes cell differentiation, and the apoptosis of colonocytes. To model a high-fiber diet, increasing the synthesis of Acetyl-CoA carboxylase, an enzyme that catabolizes glucose into SCFAs, Escherichia coli was cultured in Luria Broth enhanced with a high to low concentration of glucose. The 15mM, a high concentration of glucose, yielded qPCR products measured, for the target gene accA, which was 4,210ng/µL. The 7.5mM sample produced a concentration equaled to 375 ng/µL, and the 0µM sample measured an accA concentration of 196 ng/µL. The gene accA, 1 of 4 subunits for the Acetyl-CoA Carboxylase enzyme, was suppressed by asRNA, producing a qPCR concentration of 63ng/µL. Antisense RNA for accA reduced the amount of Lux-S, a vital gene needed for propagating quorum-sensing signal molecules. The Lux-S gene, responsible for releasing autoinducer 2 for cell to cell quorum sensing, was reduced by the gene inhibition of accA with asRNA. The increase in Lux-S transcription increases biofilm production for spreading virulence. The further implications of the study propose designing antibiotics that target bacterial cell metabolic processes to block bacterial antibiotic resistance.
Cite this as
2019. Integrating the extracellular, intracellular, and intercellular metabolic processes of Escherichia coli through glucose saturation, inhibition of the acetyl-CoA carboxylase subunit accA with asRNA, and through quantifying cell to cell quorum-sensing. PeerJ Preprints 7:e27459v2 https://doi.org/10.7287/peerj.preprints.27459v2Author comment
A co-author was added. The preprint focuses more on the gene accA than the entire microbiome of the gut. The preprint now has a more focused intent and purpose. Statistics were included where needed. Many uneeded misinterpretations were deleted. The overall preprint was revised to express a more central and single purpose.
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Competing Interests
The authors declare that they have no competing interests.
Author Contributions
Tatiana Hillman conceived and designed the experiments, performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the paper, approved the final draft.
Cory Tobin conceived and designed the experiments, analyzed the data, contributed reagents/materials/analysis tools, authored or reviewed drafts of the paper, approved the final draft.
Data Deposition
The following information was supplied regarding data availability:
The raw data included shows the effects of glucose and antisense RNA, on the production of the accA, important for translation of Acetyl-CoA Carboxylase for metabolizing glucose and for quorum cell to cell sensing.
Funding
The authors received no funding for this work.
