Matching Items (3)
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- All Subjects: Metabolism
- All Subjects: Environmental
- Genre: Doctoral Dissertation
- Creators: Torres, Cesar
Description
The accurate and fast determination of carbon dioxide (CO2) levels is critical for many health and environmental applications. For example, the analysis of CO2 levels in exhaled breath allows for the evaluation of systemic metabolism, perfusion, and ventilation, and provides the doctors and patients with a non-invasive and simple method to predict the presence and severity of asthma, and Chronic Obstructive Pulmonary Disease (COPD). Similarly, the monitoring of CO2 levels in the atmosphere allows for assessment of indoor air quality (IAQ) as the indoor CO2 levels have been proved to be associated with increased prevalence of certain mucous membrane and respiratory sick building syndrome (SBS) symptoms. A pocket-sized CO2 analyzer has been developed for real-time analysis of breath CO2 and environmental CO2. This CO2 analyzer is designed to comprise two key components including a fluidic system for efficient gas sample delivery and a colorimetric detection unit integrated into the fluidic system. The CO2 levels in the gas samples are determined by a disposable colorimetric sensor chip. The sensor chip is a novel composite based sensor that has been optimized to provide fast and reversible response to CO2 over a wide concentration range, covering the needs of both environmental and health applications. The sensor is immune to the presence of various interfering gases in ambient or expired air. The performance of the sensor in real-time breath-by-breath analysis has also been validated by a commercial CO2 detector. Furthermore, a 3D model was created to simulate fluid dynamics of breath and chemical reactions for CO2 assessment to achieve overall understanding of the breath CO2 detection process and further optimization of the device.
ContributorsZhao, Di (Author) / Forzani, Erica S (Thesis advisor) / Lin, Jerry Ys (Committee member) / Torres, Cesar (Committee member) / Tsow, Tsing (Committee member) / Xian, Xiaojun (Committee member) / Arizona State University (Publisher)
Created2014
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
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