The development of anaerobic digestion processes has gained recognition for its potential for producing volatile fatty acids alongside bio-alcohols. In addition, the fermentation process and the products produced from biological fermentation have the possibility to overcome the abundance of organic waste in our society, which is potentially immensely rich in untapped valuable potential products. The current project aims at producing medium chained volatile fatty acids and alcohols from short-chain volatile fatty acids using hydrogen as an electron donor. For efficient conversion, inhibition of methanogenic microorganisms was performed by thermally pre-treating the microorganisms at 90 °C for 15 min. The highest observed concentrations of volatile fatty acid were 6.42 ± 0.09 g/L and the concentration of ethanol was 0.33±0.03 g/L. These concentrations were obtained with the addition of 4 g/L of liquid substrate and gas co-substrate (H2:CO2). Moreover, the predominant product from the present experiment was valeric acid and it reached its highest concentration of 1.41 g/L after 37 days. Interestingly, the addition of H2:CO2 co-substrate showed that this fermentation can be used for carbon capture and utilisation alongside hydrogen consumption in a ratio of 1:1 to increase the value of short-chain fatty acids. Furthermore, this can contribute to decreased CO2 emissions and reduced use of fossil resources for alcohol production which is in line with the global environmental goals.

Perfusion operation mode is currently under fast expansion in mammalian cell based manufacturing of biopharmaceuticals, not only for labile drug protein but also for stable proteins such as monoclonal antibodies (mAbs). Perfusion mode can advantageously offer a stable cell environment, long-term production with high productivity and consistent product quality. Intensified high cell density culture (HCDC) is certainly one of the most attractive features of a perfusion process due to the high volumetric productivity in a small footprint that it can provide. Advancements in single-use technology have alleviated the intrinsic complexity of perfusion processes while the maturing in cell retention devices has improved process robustness. The knowledge for perfusion process has been gradually built and the “continuous” concept is getting more and more acceptance in the field. This thesis presents the development of robust perfusion process at very high cell densities in various culture systems. Four HCDC perfusion systems were developed with industrial collaborators with three different mAb producing Chinese Hamster Ovary (CHO) cell lines: 1-2) WAVE Bioreactor™ Cellbag prototype equipped with cell separation by hollow fiber filter utilizing Alternating Tangential Flow (ATF) and Tangential Flow Filtration (TFF) techniques; 3) Fiber matrix based CellTank™ prototype; 4) Glass stirred tank bioreactor equipped with ATF. In all the systems, extremely high viable cell densities above 130 million viable cells per milliliter (MVC/mL) up to 214 MVC/mL were achieved. Steady states were maintained and studied at 20-30 MVC/mL and 100-130 MVC/mL for process development. Perfusion rate selection based on cell specific perfusion rate (CSPR) was systematically investigated and exometabolome study was performed to explore the metabolic footprint of HCDC perfusion process. / <p>QC 20170523</p>

Biomass is a valuable and limited resource that should be used efficiently. The potential of replacing fossil-based products with bio-based ones produced in biobased industrial systems is huge. One important aim of increasing the share of biobased products is to improve the sustainability of systems for production and consumption. Therefore, it is important to evaluate what solutions are available to improve the sustainability performance of bio-based industrial systems, and if they also bring negative impacts. The thesis focuses on assessing the role of biogas solutions in developing sustainable bio-based systems. Such assessments are often quite narrow in their scope and focus on quantitative environmental or economic aspects. This thesis aims at also including feasibility related aspects involving the contextual conditions that are assessed more qualitatively. Biogas solutions are identified as a versatile approach to treat organic materials which are generated in large volumes in bio-based industrial systems. The results show that biogas solutions in bio-based industrial systems (i) improve circular flows of energy and nutrients, (ii) are especially viable alternatives when the quality of the by-product streams become poorer, and (iii) may improve the profitability of the bio-based industrial system. To perform better assessments of these systems, it seems valuable to broaden the set of indicators assessed and include feasibility-related indicators, preferably through the involvement of relevant stakeholders as they contribute with different perspectives and can identify aspects that influence the sustainability in different areas. Future studies could benefit from applying those broader assessments on more cases to build on a more generalisable knowledge base.

Material degradation and a decrease of fuel quality are common phenomena when storing biomass. A magnitude of 7.8% has been reported to degrade over five months when storing spruce wood chips in the winter in Central Europe. This thesis presents a theoretical study of biomass storage. It includes investigations of bio-chemical, chemical and physical processes that occur during storage of chipped woody biomass. These processes lead to degradation caused by micro-activity, chemical oxidation reactions and physical transformation of water. Micro-activity was modeled with Monod kinetics which are Michaelis-Menten type of expressions. The rate expressions were complemented with dependency functions describing the impact of oxygen, moisture and temperature. The woody biomass was divided into three fractions. These fractions represent how hard different components of the wood are to degrade by microorganisms. Chemical oxidation was modeled as a first order rate expression with respect to the active components of the wood. Two different cases have been simulated during the project. Firstly, an isolated system with an initial oxygen concentration of air was considered. This case displayed a temperature increase of approximately 2˚C and a material degradation less than 1%. The second case considered an isolated system with an endless depot of oxygen. This case resulted in degradation losses around 0.45-0.95% in the temperature range between 65-80˚C during approximately 300 days of storage. The temperature increased slowly due to chemical oxidation.

Since the 1900’s it has been known that microorganisms are capable of generating electrical power through extracellular electron transfer by converting the energy found organic compounds (Potter, 1911). Microbial fuel cells (MFCs) has garnered more attention recently, and have shown promise in several applications, including wastewater treatment (Yakar et al., 2018), bioremediation (Rosenbaum &amp; Franks, 2014), biosensors (ElMekawy et al., 2018) desalination (Zhang et al., 2018) and as an alternative renewable energy source in remote areas (Castro et al., 2014). In MFCs catalytic reactions of microorganisms oxidize an electron donor through extracellular electron transfer to the anode, under anaerobic conditions, with the cathode exposed to an electron acceptor, facilitating an electrical current (Zhuwei, Haoran &amp; Tingyue, 2007; Lovley, 2006). For energy production in remote areas a low cost and easily accessible feed stock is required for the MFCs. Sweet sorghum is a drought tolerant feedstock with high biomass and sugar yields, good water-use efficiency, established production systems and the potential for genetic improvements. Because of these advantages sweet sorghum stalks were proposed as an attractive feedstock (Rooney et al., 2010; Matsakas &amp; Christakopoulos, 2013). Dried sweet sorghum stalks were, therefore, tested as a raw material for power generation in a MFC, with anaerobic sludge from a biogas plant as inoculum (Sjöblom et al., 2017a). Using sorghum stalks the maximum voltage obtained was 546±10 mV, the maximum power and current density of 131±8 mW/m2 and 543±29 mA/m2 respectively and the coulombic efficiency was 2.2±0.5%. The Ohmic resistances were dominant, at an internal resistance of 182±17 Ω, calculated from polarization data. Furthermore, hydrolysis of the dried sorghum stalks did not improve the performance of the MFC but slightly increased the total energy per gram of substrate. During the MFC operation, the sugars were quickly fermented to formate, acetate, butyrate, lactate and propionate with acetate and butyrate being the key acids during electricity generation. Efficient electron transfer between the microorganisms and the electrodes is an essential aspect of bio-electrochemical systems such as microbial fuel cells. In order to design more efficient reactors and to modify microorganisms, for enhanced electricity production, understanding the mechanisms and dynamics of the electron transport chain is important. It has been found that outer membrane C-type cytochromes (OMCs) (including omcS and omcZ discussed in this study) play a key role in the electron transport chain of Geobacter sulfurreducens, a well-known, biofilm forming, electro-active microorganism  (Millo et al., 2011; Lovley, 2008). It was found that Raman microscopy is capable of providing biochemical information, i.e., the redox state of c-type cytochromes (cyt-C) without damaging the microbial biofilm, allowing for in-situ observation. Raman microscopy was used to observe the oxidation state of OMCs in a suspended culture, as well as in a biofilm of an MFC. First, the oxidation state of the OMCs of suspended cultures from three G. sulfurreducens strains (PCA, KN400 and ΔpilA) was analyzed. It was found that the oxidation state can also be used as an indicator of the metabolic state of the cells, and it was confirmed that PilA, a structural pilin protein essential for long range electron transfer, is not required for external electron transfer. Furthermore, we designed a continuous, anaerobic MFC enabling in-situ Raman measurements of G. sulfurreducens biofilms during electricity generation, while poised using a potentiostat, in order to monitor and characterize the biofilm. Two strains were used, a wild strain, PCA, and a mutant, ΔOmcS. The cytochrome redox state, observed through the Raman spectra, could be altered by applying different poise voltages to the electrodes. This change was indirectly proportional to the modulation of current transferred from the cytochromes to the electrode. This change in Raman peak area was reproducible and reversible, indicating that the system could be used, in-situ, to analyze the oxidation state of proteins responsible for the electron transfer process and the kinetics thereof.

Microalgae are considered as a potential feedstock for many promising applications. Some active substances in microalgae have plant biostimulation effects potentially useful in agriculture. However, to produce such a microalgal biomass, specific microalgae cultivation and post-treatment processes must be designed to preserve active substances. A particular focus is provided on cultivation (tubular photobioreactor) and different plausible post-treatment scenarios for microalgae separation (flocculation and centrifugation) and preservation (sterilization and drying). For each step, yield and energy consumption are modeled using data taken from literature or lab and pilot scale experiments. Industrial equipment for scale-up process is also studied by comparing existing systems. These models enable to make an economic evaluation of the whole process and to study its profitability for each scenario. The breakeven price is calculated as a function of the production rate. Several parameters are suggested to improve system efficiency and profitability at the end of this study. However, a better microalgae characterization and more experiments on potential post-treatment systems are required to improve the accuracy of the model.

One of the main interests in commercial methane production is to maximize the gas yield, and it is thus appealing to use material with relative high methane potential. However, such material often results in process instability whereas ammonia inhibition is common. Removal of ammonia through adsorption is a fairly unexplored method in the field of biogas production, and could prove to be cost-effective.The adsorption capacity of pulp fibres from the paper making industry were investigated through batch adsorption experiments. Additionally, the fibres effect on small scale batch digesters in terms of methane production and cellulase activity was explored. Overall, the adsorption capacity of the pulp fibres was low, whereas Kraft hardwood had the highest adsorption capacity in both an aqueous ammonium solution and digester fluid at 11±3 and 60±20 mg g-1, respectively. The initial total ammonium nitrogen concentration had the highest effect on the adsorption capacity with a positive correlation. The pulp fibres seemingly had no effect on the ammonia inhibited anaerobic digestion systems. However, the cellulase activity was higher after day 5 in the anaerobic digestion systems with a high ammonia concentration.In essence, the overall results showed that the adsorption of the fibres was relatively low and most likely not suitable as a material to prevent ammonia inhibition in an AD.

Food waste and fashion pollution are two of the significant global environmental issues throughout the recent past. In this research, it was investigated the feasibility of making a leather-like material from bread waste using biotechnology as the bridging mechanism. The waste bread collected from the supermarkets were used as the substrate to grow filamentous fungi species Rhizopus Delemar and Fusarium Venenatum. Tanning of fungal protein fibres was successfully performed using vegetable tanning, confirmed using FTIR and SEM images. Furthermore, glycerol and a biobased binder treatment was performed for the wet-laid fungal microfibre sheets produced. Overall, three potential materials were able to produce with tensile strengths ranging from 7.74 ± 0.55 MPa to 6.92 ± 0.51 MPa and the elongation% from 16.81 ± 1.61 to 4.82 ± 0.36. The binder treatment enhanced the hydrophobicity even after the glycerol treatment, an added functional advantage for retaining flexibility even after contact with moisture. The fungal functional material produced with bread waste can be tailored successfully into leather substitutes using an environmentally benign procedure.

Syftet med projektet är att undersöka effekten av att leda över vattenverksslam från Norrvattens vattenverk, Görvälnverket, till Käppalaverkets reningsverk som ett steg i en mer hållbar slamhantering. Detta slamhanterings alternativ jämförs med en framtida lokal slamhantering vid Görvälnverket. Studien undersöker möjligheten att leda vattenverksslam över till Käppalaverket utifrån fyra huvudaspekter, vattenrening, drift, kostnad och miljöpåverkan. En litteraturstudie genomfördes med syfte att utvärdera hur vattenverksslammet kan påverka reningsprocesserna vid Käppalaverket. En Livscykelanalys genomfördes med syfte att utvärdera miljöpåverkan av att leda över vattenverksslam till Käppalaverket. Genomförbarheten utvärderades med hjälp av en multikriterieanalys, där tekniska, miljömässiga och ekonomiska aspekter utvärderades.  Resultat från studien visade att den framtida lokala slamhanteringen är mer fördelaktig från ett tekniskt och ekonomiskt perspektiv, än överledning av vattenverksslammet till Käppalaverket. En nackdel med överledning av vattenverksslam till Käppalaverket är att vattenverksslammet sannolikt kommer påverka avvattningen av avloppsslammet, vilket resulterar i en högre polymerförbrukning och en ökad hydraulisk belastning på centrifuger och rötkammare. Överledningen av vattenverksslam antas däremot inte ha en negativ påverka på kvaliteten av reningen vid Käppalaverket. Att leda vattenverksslam över till Käppalaverket ger en lägre miljöpåverkan med avseende på kemikalieutsläpp till vattenmiljön men en högre miljöpåverkan med avseende på transporter och energiförbrukning.  Överledning av vattenverksslam till Käppalaverket bedöms som genomförbart, men denna studie visar att den framtida lokala slamhanteringen på Görvälnverket är ett mer fördelaktigt alternativ. Om överledning av vattenverksslam till Käppalaverket fortsatt är ett aktuellt alternativ rekommenderas det att en mer djupgående studie utförs, där vattenverksslammet tillsätts till Käppalaverket för att utvärdera dess påverkan. / The aim with this project is to investigate the impact of leading the produced waterworks sludge (WWS) from Norrvattens drinking water treatment plant (DWTP), Görvälnverket, over to Käppalaverkets wastewater treatment plant (WWTP) as a step in a more sustainable sludge management. This alternative is compared to a future sludge management at Görvälnverket. The study, investigating the feasibility of leading the WWS over to Käppalaverket, is based on four main aspects, water treatment, operation, cost, and environmental impact. A literature study was performed to evaluate the effect of WWS on Käppalaverket. A life cycle assessment (LCA) analysis was performed to evaluate the environmental impact of leading the WWS over to Käppalaverket. The feasibility was evaluated using a multi-criteria decision analysis (MCDA), where technical, environmental, and economic aspects were considered. The study shows that the future sludge management is more favourable than leading the WWS over to Käppalaverket from a technical and economic aspect. The main drawback with leading the WWS over to Käppalaverket is that the WWS will likely impact the dewatering of the sewage sludge, resulting in a higher polymer consumption and an increased hydraulic load on centrifuges and digesters. However, the addition of WWS at Käppalaverket is not assumed to negatively impact the quality of the treatment at Käppalaverket. Leading the WWS over to Käppalaverket results in a lower environmental impact regarding chemical emissions but results in a higher environmental impact regarding transportation and energy consumption. Leading the WWS over to Käppalaverket was found to be feasible, although the future sludge management at Görvälnverket was found to be more favourably in this study. A more in depth study on the feasibility of leading the WWS over to Käppalaverkets is required to fully assess this aspect. A trial where the WWS is added to Käppalaverket is recommended to further evaluate the impact of the WWS.

To diagnose patients suffering from blood clotting disorders latex immununoassays (LIA) can be used. A time consuming manual tangential flow filtration (TFF) process suggests the implementation of an automatic TFF system to improve the efficiency, profitability, and expandability of the production facility of LIA at Nordic Biomarker. Tests were made of the automatic TFF system's ability to perform the desired steps of concentration, dilution and diafiltration, both with purified water and mimicked product. The mimicked product of micro particles (MP) mixed with monoclonal antibodies (mAb) was also used to further test the system's pressure control, safety alarms and stops, and to determine a permeate flux by a critical flux experiment. The results imply a functional TFF system able to automatically concentrate the process fluid and maintain a stable volume during diafiltration, although an additional permeate pump was ordered to be able to attain a fully functional performance of the automatic TFF process. The final part of the implementation was to initiate a validation draft including a risk assessment, OQ plan and PQ plan that resulted in a plan of the main tests to be performed. To conclude, the essential part of the implementation of a high quality and efficient automatic TFF process was conducted to facilitate future expansion of the production of LIA.