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"Biofilmomics": functional protein expression in biofilm biotechnologies revealed by quantitative proteomics

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dc.contributor.authorChignell, Jeremy, author
dc.contributor.authorReardon, Kenneth, advisor
dc.contributor.authorDe Long, Susan, advisor
dc.contributor.authorPeebles, Christie, committee member
dc.contributor.authorSharvelle, Sybil, committee member
dc.date.accessioned2020-06-22T11:54:01Z
dc.date.available2022-06-15T11:54:01Z
dc.date.issued2020
dc.description.abstractMicrobial biotechnologies that utilize biofilms often exhibit superior performance compared with planktonic systems. Many details of biofilm metabolism that drive those improvements in performance remain unclear. Only recently have molecular tools emerged that can provide a holistic picture of life in a complex biosystem like a biofilm for the purposes of answering questions on a system level. The purpose of this work was to address four fundamental questions about protein expression in biofilms: what kind of protein expression is distinctive to biofilms? Which biofilm proteins are associated with a function of interest? How does co-culture with another species affect biofilm-related protein expression? When during multi-species biofilm development does a function of interest emerge and who in the community is responsible? Label-free quantitative proteomics was used in conjunction with physiological experimentation to address these four questions. In the first study we found that L. delbrueckii lactis protein expression in flow-cell biofilms was 31% more diverse than in planktonic cultures, and proteins related to catalytic activity were significantly increased in biofilms at the expense of proteins for cell motility and replication. Roles for riboflavin and fatty acid metabolism suggested modulations in redox functions and membrane turnover during life in a biofilm. The second study compared protein expression by S. onedensis MR-1 in electricity-generating biofilms with that in aerobic biofilms from the same microbial fuel cell reactor. Three novel proteins associated with electricity generation were identified, in addition to proteomic evidence of aerobic metabolism by anode biofilm cells. The latter result was shown to be consistent with kinetics of oxygen depletion and bulk cell growth in the MFC, suggesting operational conditions to reduce this bulk cell growth and thereby reduce fouling of the cathode and improve overall Coulombic efficiency of the single-chamber MFC system. In the third study, it was discovered through proteomic and physiological experiments that a virulent phenotype associated with biofilm formation was triggered in P. putida when co-cultured with B. atrophaeus. Dramatic shifts in protein expression at the initial trigger point of virulent biofilm formation by P. putida are described. Finally, a comparison of the meta-proteomes of microbial fuel cell biofilms at different stages of development indicated that proteins in metabolic pathways for carbon storage and competitive inhibition are differentially expressed when the biofilm becomes electrochemically active. Meta-proteomics and 16S rRNA gene sequencing agreed that it is possible for a microbial fuel cell community to maintain high diversity (and therefore potentially higher resilience) while generating electricity at levels comparable to a MFC community dominated by Geobacter. Each of these chapters was prepared as an independent manuscript, though the themes were integrated by the overall theme of quantifying differential protein expression in biofilms in order to reveal new details about their development and functionality. Since the performance of many engineered biosystems—including those that employ biofilms—often can be controlled adequately at an operational level, an attitude persists that any additional molecular investigation is superfluous. The work presented here provides evidence for the opposite viewpoint: a rich understanding of the molecular mechanisms behind biofilm functionality can inform strategies for continuous system improvement and suggest new capabilities and biotechnological applications of biofilms.
dc.format.mediumborn digital
dc.format.mediumdoctoral dissertations
dc.identifierChignell_colostate_0053A_16046.pdf
dc.identifier.urihttps://hdl.handle.net/10217/208595
dc.languageEnglish
dc.language.isoeng
dc.publisherColorado State University. Libraries
dc.relation.ispartof2020-
dc.rightsCopyright and other restrictions may apply. User is responsible for compliance with all applicable laws. For information about copyright law, please see https://libguides.colostate.edu/copyright.
dc.subjectbiofilm
dc.subjectlactic
dc.subjectproteomics
dc.subjectengineering
dc.subjectbacteria
dc.subjectmicrobial fuel cell
dc.title"Biofilmomics": functional protein expression in biofilm biotechnologies revealed by quantitative proteomics
dc.title.alternativeBiofilmomics: functional protein expression in biofilm biotechnologies revealed by quantitative proteomics
dc.typeText
dcterms.embargo.expires2022-06-15
dcterms.embargo.terms2022-06-15
dcterms.rights.dplaThis Item is protected by copyright and/or related rights (https://rightsstatements.org/vocab/InC/1.0/). You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s).
thesis.degree.disciplineChemical and Biological Engineering
thesis.degree.grantorColorado State University
thesis.degree.levelDoctoral
thesis.degree.nameDoctor of Philosophy (Ph.D.)

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