Matching Items (3)
Filtering by
- All Subjects: Metabolism
- All Subjects: Adaptive control
- Genre: Academic theses
- Creators: Torres, Cesar
Description
Metabolic engineering is an extremely useful tool enabling the biosynthetic production of commodity chemicals (typically derived from petroleum) from renewable resources. In this work, a pathway for the biosynthesis of styrene (a plastics monomer) has been engineered in Escherichia coli from glucose by utilizing the pathway for the naturally occurring amino acid phenylalanine, the precursor to styrene. Styrene production was accomplished using an E. coli phenylalanine overproducer, E. coli NST74, and over-expression of PAL2 from Arabidopsis thaliana and FDC1 from Saccharomyces cerevisiae. The styrene pathway was then extended by just one enzyme to either (S)-styrene oxide (StyAB from Pseudomonas putida S12) or (R)-1,2-phenylethanediol (NahAaAbAcAd from Pseudomonas sp. NCIB 9816-4) which are both used in pharmaceutical production. Overall, these pathways suffered from limitations due to product toxicity as well as limited precursor availability. In an effort to overcome the toxicity threshold, the styrene pathway was transferred to a yeast host with a higher toxicity limit. First, Saccharomyces cerevisiae BY4741 was engineered to overproduce phenylalanine. Next, PAL2 (the only enzyme needed to complete the styrene pathway) was then expressed in the BY4741 phenylalanine overproducer. Further strain improvements included the deletion of the phenylpyruvate decarboxylase (ARO10) and expression of a feedback-resistant choristmate mutase (ARO4K229L). These works have successfully demonstrated the possibility of utilizing microorganisms as cellular factories for the production styrene, (S)-styrene oxide, and (R)-1,2-phenylethanediol.
ContributorsMcKenna, Rebekah (Author) / Nielsen, David R (Thesis advisor) / Torres, Cesar (Committee member) / Caplan, Michael (Committee member) / Jarboe, Laura (Committee member) / Haynes, Karmella (Committee member) / Arizona State University (Publisher)
Created2014
Description
Microbial fuel cells(MFC) use micro-organisms called anode-respiring bacteria(ARB) to convert chemical energy into electrical energy. This process can not only treat wastewater but can also produce useful byproduct hydrogen peroxide(H2O2). Process variables like anode potential and pH play important role in the MFC operation and the focus of this dissertation are pH and potential control problems.
Most of the adaptive pH control solutions use signal-based-norms as cost functions, but their strong dependency on excitation signal properties makes them sensitive to noise, disturbances, and modeling errors. System-based-norm( H-infinity) cost functions provide a viable alternative for the adaptation as they are less susceptible to the signal properties. Two variants of adaptive pH control algorithms that use approximate H-infinity frequency loop-shaping (FLS) cost metrics are proposed in this dissertation.
A pH neutralization process with high retention time is studied using lab scale experiments and the experimental setup is used as a basis to develop a first-principles model. The analysis of such a model shows that only the gain of the process varies significantly with operating conditions and with buffering capacity. Consequently, the adaptation of the controller gain (single parameter) is sufficient to compensate for the variation in process gain and the focus of the proposed algorithms is the adaptation of the PI controller gain. Computer simulations and lab-scale experiments are used to study tracking, disturbance rejection and adaptation performance of these algorithms under different excitation conditions. Results show the proposed algorithm produces optimum that is less dependent on the excitation as compared to a commonly used L2 cost function based algorithm and tracks set-points reasonably well under practical conditions. The proposed direct pH control algorithm is integrated with the combined activated sludge anaerobic digestion model (CASADM) of an MFC and it is shown pH control improves its performance.
Analytical grade potentiostats are commonly used in MFC potential control, but, their high cost (>$6000) and large size, make them nonviable for the field usage. This dissertation proposes an alternate low-cost($200) portable potentiostat solution. This potentiostat is tested using a ferricyanide reactor and results show it produces performance close to an analytical grade potentiostat.
Most of the adaptive pH control solutions use signal-based-norms as cost functions, but their strong dependency on excitation signal properties makes them sensitive to noise, disturbances, and modeling errors. System-based-norm( H-infinity) cost functions provide a viable alternative for the adaptation as they are less susceptible to the signal properties. Two variants of adaptive pH control algorithms that use approximate H-infinity frequency loop-shaping (FLS) cost metrics are proposed in this dissertation.
A pH neutralization process with high retention time is studied using lab scale experiments and the experimental setup is used as a basis to develop a first-principles model. The analysis of such a model shows that only the gain of the process varies significantly with operating conditions and with buffering capacity. Consequently, the adaptation of the controller gain (single parameter) is sufficient to compensate for the variation in process gain and the focus of the proposed algorithms is the adaptation of the PI controller gain. Computer simulations and lab-scale experiments are used to study tracking, disturbance rejection and adaptation performance of these algorithms under different excitation conditions. Results show the proposed algorithm produces optimum that is less dependent on the excitation as compared to a commonly used L2 cost function based algorithm and tracks set-points reasonably well under practical conditions. The proposed direct pH control algorithm is integrated with the combined activated sludge anaerobic digestion model (CASADM) of an MFC and it is shown pH control improves its performance.
Analytical grade potentiostats are commonly used in MFC potential control, but, their high cost (>$6000) and large size, make them nonviable for the field usage. This dissertation proposes an alternate low-cost($200) portable potentiostat solution. This potentiostat is tested using a ferricyanide reactor and results show it produces performance close to an analytical grade potentiostat.
ContributorsJoshi, Rakesh (Author) / Tsakalis, Konstantinos (Thesis advisor) / Rodriguez, Armando (Committee member) / Torres, Cesar (Committee member) / Spanias, Andreas (Committee member) / Arizona State University (Publisher)
Created2018
Description
This dissertation focuses on the biosynthetic production of aromatic fine chemicals in engineered Escherichia coli from renewable resources. The discussed metabolic pathways take advantage of key metabolites in the shikimic acid pathway, which is responsible for the production of the aromatic amino acids phenylalanine, tyrosine, and tryptophan. For the first time, the renewable production of benzaldehyde and benzyl alcohol has been achieved in recombinant E. coli with a maximum titer of 114 mg/L of benzyl alcohol. Further strain development to knockout endogenous alcohol dehydrogenase has reduced the in vivo degradation of benzaldehyde by 9-fold, representing an improved host for the future production of benzaldehyde as a sole product. In addition, a novel alternative pathway for the production of protocatechuate (PCA) and catechol from the endogenous metabolite chorismate is demonstrated. Titers for PCA and catechol were achieved at 454 mg/L and 630 mg/L, respectively. To explore potential routes for improved aromatic product yields, an in silico model using elementary mode analysis was developed. From the model, stoichiometric optimums maximizing both product-to-substrate and biomass-to-substrate yields were discovered in a co-fed model using glycerol and D-xylose as the carbon substrates for the biosynthetic production of catechol. Overall, the work presented in this dissertation highlights contributions to the field of metabolic engineering through novel pathway design for the biosynthesis of industrially relevant aromatic fine chemicals and the use of in silico modelling to identify novel approaches to increasing aromatic product yields.
ContributorsPugh, Shawn (Author) / Nielsen, David (Thesis advisor) / Dai, Lenore (Committee member) / Torres, Cesar (Committee member) / Lind, Mary Laura (Committee member) / Wang, Xuan (Committee member) / Arizona State University (Publisher)
Created2016