Modeling and optimization of extracellular polysaccharides production by Enterobacter A47

Polysaccharides are gaining increasing attention as potential environmental friendly and sustainable building blocks in many fields of the (bio)chemical industry. The microbial production of polysaccharides is envisioned as a promising path, since higher biomass growth rates are possible and therefore higher productivities may be achieved compared to vegetable or animal polysaccharides sources. This Ph.D. thesis focuses on the modeling and optimization of a particular microbial polysaccharide, namely the production of extracellular polysaccharides (EPS) by the bacterial strain Enterobacter A47. Enterobacter A47 was found to be a metabolically versatile organism in terms of its adaptability to complex media, notably capable of achieving high growth rates in media containing glycerol byproduct from the biodiesel industry. However, the industrial implementation of this production process is still hampered due to a largely unoptimized process. Kinetic rates from the bioreactor operation are heavily dependent on operational parameters such as temperature, pH, stirring and aeration rate. The increase of culture broth viscosity is a common feature of this culture and has a major impact on the overall performance. This fact complicates the mathematical modeling of the process, limiting the possibility to understand, control and optimize productivity. In order to tackle this difficulty, data-driven mathematical methodologies such as Artificial Neural Networks can be employed to incorporate additional process data to complement the known mathematical description of the fermentation kinetics. In this Ph.D. thesis, we have adopted such an hybrid modeling framework that enabled the incorporation of temperature, pH and viscosity effects on the fermentation kinetics in order to improve the dynamical modeling and optimization of the process. A model-based optimization method was implemented that enabled to design bioreactor optimal control strategies in the sense of EPS productivity maximization. It is also critical to understand EPS synthesis at the level of the bacterial metabolism, since the production of EPS is a tightly regulated process. Methods of pathway analysis provide a means to unravel the fundamental pathways and their controls in bioprocesses. In the present Ph.D. thesis, a novel methodology called Principal Elementary Mode Analysis (PEMA) was developed and implemented that enabled to identify which cellular fluxes are activated under different conditions of temperature and pH. It is shown that differences in these two parameters affect the chemical composition of EPS, hence they are critical for the regulation of the product synthesis. In future studies, the knowledge provided by PEMA could foster the development of metabolically meaningful control strategies that target the EPS sugar content and oder product quality parameters.

Sustainable urban planning: parameters to the operational process

In modern society, energy consumption and respect for the environment have become essential aspects of urban planning. The rising demand for alternative sources of energy, coupled with the decline in the construction sector and material usage, gives the idea that the thinking on modern cities, where attention is given to reduced energy consumption, savings, waste recycling and respect for the surrounding environment, is being put into practice. If we examine development of the city over recent centuries, by means of the theories of the most famous and influential urban planners, it is possible to identify the major problems caused by this type of planning. For this reason, in recent urban planning the use of systems of indicators that evaluate and certify land environmentally and energetically guides the master plan toward a more efficient city model. In addition the indicators are targeted on key factors determined by the commissioner or the opportunities the territory itself provides. Due the complexity of the environmental mechanics, the process of design and urban planning has become a challenging issue. The introduction of the indicators system has made it possible to register the life of the process, with a spiral route that allows the design itself to be refined. The aim of this study, built around the creation of a system of urban sustainability indicators that will evaluate highly eco-friendly cities, is to develop a certification system for cities or portions of them. The system will be upgradeable and objective, will employ real data and will be concerned with energy production and consumption.

The effect of key process operational conditions on enhanced biological phosphorus removal from wastewater

Enhanced biological phosphorus removal (EBPR) is the most economic and sustainable option used in wastewater treatment plants (WWTPs) for phosphorus removal. In this process it is important to control the competition between polyphosphate accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs), since EBPR deterioration or failure can be related with the proliferation of GAOs over PAOs. This thesis is focused on the effect of operational conditions (volatile fatty acid (VFA) composition, dissolved oxygen (DO) concentration and organic carbon loading) on PAO and GAO metabolism. The knowledge about the effect of these operational conditions on EBPR metabolism is very important, since they represent key factors that impact WWTPs performance and sustainability. Substrate competition between the anaerobic uptake of acetate and propionate (the main VFAs present in WWTPs) was shown in this work to be a relevant factor affecting PAO metabolism, and a metabolic model was developed that successfully describes this effect. Interestingly, the aerobic metabolism of PAOs was not affected by different VFA compositions, since the aerobic kinetic parameters for phosphorus uptake, polyhydroxyalkanoates (PHAs) degradation and glycogen production were relatively independent of acetate or propionate concentration. This is very relevant for WWTPs, since it will simplify the calibration procedure for metabolic models, facilitating their use for full-scale systems. The DO concentration and aerobic hydraulic retention time (HRT) affected the PAO-GAO competition, where low DO levels or lower aerobic HRT was more favourable for PAOs than GAOs. Indeed, the oxygen affinity coefficient was significantly higher for GAOs than PAOs, showing that PAOs were far superior at scavenging for the often limited oxygen levels in WWTPs. The operation of WWTPs with low aeration is of high importance for full-scale systems, since it decreases the energetic costs and can potentially improve WWTP sustainability. Extended periods of low organic carbon load, which are the most common conditions that exist in full-scale WWTPs, also had an impact on PAO and GAO activity. GAOs exhibited a substantially higher biomass decay rate as compared to PAOs under these conditions, which revealed a higher survival capacity for PAOs, representing an advantage for PAOs in EBPR processes. This superior survival capacity of PAOs under conditions more closely resembling a full-scale environment was linked with their ability to maintain a residual level of PHA reserves for longer than GAOs, providing them with an effective energy source for aerobic maintenance processes. Overall, this work shows that each of these key operational conditions play an important role in the PAO-GAO competition and should be considered in WWTP models in order to improve EBPR processes.