Chemical and Biomolecular Engineering Theses and Dissertations
Permanent URI for this collectionhttp://hdl.handle.net/1903/2751
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Item Evaluation of the transcription of small RNA SgrS and glucose transporter mRNA ptsG in E. coli B and E. coli K cultures under high glucose conditions(2009) Ng, Weng Ian; Wang, Nam Sun; Chemical Engineering; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Escherichia coli is commonly used as the production system for recombinant proteins. However, acetate accumulation in fermentation affects cell growth and protein yield. Recent studies have showed that the small RNA SgrS regulates the major glucose transporter mRNA ptsG in a post–transcriptional manner when the metabolic intermediate glucose–6–phosphate is accumulated intracellularly in E. coli K. Here, comparative analysis of the transcription of SgrS and ptsG is performed between E. coli B and E. coli K cultures in both shake flasks and bioreactor. Both strains expressed SgrS when grown on the non–metabolizable glucose analog α–methyl–glucoside. However, under high glucose conditions, only E. coli B showed significant expression of SgrS. This behavior is unaffected by oxygen supply and pH control. E. coli B produced less acetate on glucose than E. coli K in the bioreactor settings. This provides evidence of a possible connection between SgrS and acetate production in aerobic fermentation of E. coli.Item Inhibitors of Autoinducer-2 Quorum Sensing and Their Effect on Bacterial Biofilm Formation(2007-07-31) Lennen, Rebecca Melissa; Bentley, William E; Chemical Engineering; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Bacteria utilize small signaling molecules, or autoinducers, to regulate their gene expression in tandem by a process termed quorum sensing. The gene encoding the synthase for autoinducer-2 (AI-2), luxS, is conserved in dozens of diverse bacteria. Behaviors controlled by AI-2 include virulence, motility, toxin production, and biofilm formation. The development of therapies that interfere with AI-2 quorum sensing are attractive for targeting biofilms, which exhibit inherent resistance to most antibiotics and biocidal agents. In this study, in vitro synthesized AI-2, LuxS inhibitors, and (5Z)-4-bromo-5-(bromomethylene)-3-butyl-2(5H)-furanone were screened for their effect on biofilm formation in Escherichia coli, Bacillus cereus, and Listeria innocua. The LuxS inhibitors were found to have no influence on biofilm formation in any of the screened species, but reduced exponential phase AI-2 production in Listeria innocua. The brominated furanone significantly inhibited growth in B. cereus and L. innocua, and under certain conditions preferentially inhibited biofilm formation independently from growth.