Treatment of oily wastewater by ultrafiltration: The effect of different operating and solution conditions

In many industries, such as petroleum production, and the petrochemical, metal, food and cosmetics industries, wastewaters containing an emulsion of oil in water are often produced. The emulsions consist of water (up to 90%), oils (mineral, animal, vegetable and synthetic), surfactants and other contaminates. In view of its toxic nature and its deleterious effects on the surrounding environment (soil, water) such wastewater needs to be treated before release into natural water ways. Membrane-based processes have successfully been applied in industrial applications and are considered as possible candidates for the treatment of oily wastewaters. Easy operation, lower cost, and in some cases, the ability to reduce contaminants below existing pollution limits are the main advantages of these systems. The main drawback of membranes is flux decline due tofouling and concentration polarisation. The complexity of oil-containing systems demands complementary studies on issues related to the mitigation of fouling and concentration polarisation in membranebased ultrafiltration. In this thesis the effect of different operating conditions (factors) on ultrafiltration of oily water is studied. Important factors are normally correlated and, therefore, their effect should be studied simultaneously. This work uses a novel approach to study different operating conditions, like pressure, flow velocity, and temperature, and solution properties, like oil concentration (cutting oil, diesel, kerosene), pH, and salt concentration (CaCl2 and NaCl)) in the ultrafiltration of oily water, simultaneously and in a systematic way using an experimental design approach. A hypothesis is developed to describe the interaction between the oil drops, salt and the membrane surface. The optimum conditions for ultrafiltration and the contribution of each factor in the ultrafiltration of oily water are evaluated. It is found that the effect on permeate flux of the various factors studied strongly depended on the type of oil, the type of membrane and the amount of salts. The thesis demonstrates that a system containing oil is very complex, and that fouling and flux decline can be observed even at very low pressures. This means that only the weak form of the critical flux exists for such systems. The cleaning of the fouled membranes and the influence of different parameters (flow velocity, temperature, time, pressure, and chemical concentration (SDS, NaOH)) were evaluated in this study. It was observed that fouling, and consequently cleaning, behaved differently for the studied membranes. Of the membranes studied, the membrane with the lowest propensity for fouling and the most easily cleaned was the regenerated cellulose membrane (C100H). In order to get more information about the interaction between the membrane and the components of the emulsion, a streaming potential study was performed on the membrane. The experiments were carried out at different pH and oil concentration. It was seen that oily water changed the surface charge of the membrane significantly. The surface charge and the streaming potential during different stages of filtration were measured and analysed being a new method for fouling of oil in this thesis. The surface charge varied in different stages of filtration. It was found that the surface charge of a cleaned membrane was not the same as initially; however, the permeability was equal to that of a virgin membrane. The effect of filtration mode was studied by performing the filtration in both cross-flow and deadend mode. The effect of salt on performance was considered in both studies. It was found that salt decreased the permeate flux even at low concentration. To test the effect of hydrophilicity change, the commercial membranes used in this thesis were modified by grafting (PNIPAAm) on their surfaces. A new technique (corona treatment) was used for this modification. The effect of modification on permeate flux and retention was evaluated. The modified membranes changed their pore size around 33oC resulting in different retention and permeability. The obtained results in this thesis can be applied to optimise the operation of a membrane plant under normal or shock conditions or to modify the process such that it becomes more efficient or effective.

Palvelutasopoikkeamariskin kustannusvaikutusten määrittäminen tietoliikennepalveluiden tuotannossa

Tutkimuksen päämääränä oli selvittää laadullisin menetelmin millaisia kustannusvaikutuksia laajavaikutteiset palvelutasopoikkeamat, eli erilaiset viat tai häiriöt tietoliikennepalveluissa aiheuttavat kohdeyrityksen tietoliikennepalveluiden tuotannossa. Tutkimus muistutti lähtökohdiltaan paljolti riskianalyysia, sillä sen tavoitteina oli tunnistaa merkittävimmät palvelutasopoikkeamien aiheuttajat, arvioida palvelutasopoikkeaman kustannusvaikutuksia ja pohtia millaisin toimin ja kustannuksin palvelutasopoikkeamia voitaisiin ehkäistä. Tutkimusongelmaa lähestyttiin jakamalla palvelutasopoikkeaman kustannusvaikutukset viankorjauksesta aiheutuviin välittömiin ja asiakasvaikutusten kautta mahdollisesti syntyviin välillisiin kustannuksiin. Välillisiä kustannuksia aiheuttaviksi ilmiöiksi tunnistettiin palvelukatkon aiheuttama liikennetulomenetys, nykyisten tai potentiaalisten asiakkaiden menettäminen ja nykyisten asiakkaiden alentunut ostohalukkuus. Yhteenvetona tutkimuksessa tehtyihin havaintoihin perustuen välilliset kustannukset eivät osoittautuneet niin merkittäviksi kuin tutkimuksen aloitusvaiheessa oletettiin, mutta lisätutkimus aiheesta voi olla perusteltua.

The effectiveness of vocationally oriented multidisciplinary rehabilitation (ASLAK®) amongst public sector employees

In Finland, vocationally oriented medical rehabilitation (ASLAK®) is a common preventive rehabilitation measure with the primary goal of preserving and improving work ability. The ASLAK® programme has been used for almost 30 years, although limited data exist on its effectiveness. The aims of this study were to determine whether the increased risk of work disability predicts the participants’ likelihood to be granted ASLAK® rehabilitation and to assess the effectiveness of the programme in decreasing the risk of work disability and modifying health-risk behaviours. This study is a part of the on-going Finnish Public Sector Study conducted by the Finnish Institute of Occupational Health. Data on 53 416 employees (81% women) were gathered from employers’ records, national health registers and repeated survey responses. During the 5-year follow-up, increased levels of the risk factors for work disability did not predict participation in the rehabilitation programme. During the 2.8-year followup (range 0.04–5.0 years), the risk of long-term work disability (sick leave >90 days or retirement) overall or, more specifically, due to musculoskeletal or mental diseases did not differ between the rehabilitants who participated in ASLAK® in 1997–2005 and their propensity score matched controls. There was no evidence of ASLAK® being effective in changing participants’ health-risk behaviours or in improving perceived general or mental health. The results suggest that potential participant recognition, mainly taking place in occupational health care, may fail to identify those with a higher risk of work disability. No evidence on the effectiveness of the programme was found in the study cohort when measured by the selected indicators.

Finnish cleantech SMEs in China: Challenges and solutions

The rapid economic growth in China has resulted in environmental challenges ranging from air pollution to water-related issues. Thus supporting clean technology, or cleantech, that encompasses industries that focus on alternative energy, pollution and recycling, power supplies and conservation has become one of the focal points in the Chinese economic policy for the next decade. Simultaneously, the Finnish government has initiated programs to support the internationalisation of domestic cleantech companies in an attempt to spiral the industry into one of the pillars of Finnish economic growth. This study concentrates on the conjunction of these two themes and studies the challenges faced by Finnish cleantech SMEs in the Chinese market. Consequently, the study answers the following sub questions: 1. What human and financial resource-based challenges do Finnish cleantech SMEs face in the Chinese market and what are their solutions? 2. What knowledge-based challenges do Finnish cleantech SMEs face in the Chinese market and how can these difficulties be resolved? 3. What network-based challenges do Finnish cleantech SMEs face in the Chinese market, how do they relate to the resource- and knowledge-based challenges, and how can these difficulties be resolved? This qualitative study is conducted by analysing four semi structured interviews collected from four Finnish SMEs that operate in China. The findings of the study indicate that in human resources the most important challenges are related to the hiring and retaining of employees. In contrast to extant academic literature results distinguish salary and social status as the main solutions to this challenge. Regarding financial resources it is discovered that cleantech companies enjoy a benign business environment in China and benefit from the Chinese government’s support for cleantech industry. Challenges related to knowledge resources can be grouped into categories with the most interesting knowledge flows being the stream of local market knowledge into to the foreign parent company and the outward flow of manufacturing and business practice information into the target venture. The challenge related to the first flow is gathering relevant information and the main solutions are clustering at the foreign location and hiring knowledge prior to internationalisation. Regarding the second flow the main challenge is related to intellectual property rights and the most interesting solution is the purposeful transformation of explicit knowledge into tacit knowledge. Finally, it is discovered that networks, called guanxi in China, greatly affect the business processes. Within the guanxi system there is the concept of face which was found to affect employee propensity to stay as well as, as a novel academic result, employees’ knowledge sharing intention.

Entrepreneurial orientation in small and medium-sized enterprises during economic crisis

The economic importance of small and medium-sized enterprises (SMEs) and entrepreneurship has increased significantly in recent decades and entrepreneurial activity and SMEs are deemed vital to economic progress. Therefore, it is justifiable to study how small firms and entrepreneurs can enhance their performance and emergence in the turbulent economic environment. The concept of entrepreneurial orientation (EO) has recently attracted considerable attention in the field of entrepreneurship research. EO generally refers to a firm’s propensity to be innovative, to be proactive and to take risks. A majority of EO studies so far have found that adopting EO associated entrepreneurial behaviors will help firms to create or sustain a high level of performance. This dissertation explores the main drivers and performance implications of EO for SMEs in time of economic crisis. Hence the first objective of this dissertation is to examine the performance implications of EO and to test the role of EO on how firms are treated by the crisis at operative level. The second objective is to expand the prevailing understanding of determinants of EO by exploring the relationship between owner's work related values, attitudes, demographic characteristics, firm’s financial resources and EO. EO was found to be a significant and positive factor behind a firm’s long run growth. Hence it can be said that EO has positive implications for firm performance. But on the other hand, during a time of economic crisis the different dimensions of EO had both positive and negative effects on performance of SMEs. The performance implications varied across different stages of the crisis and were also dependent on what measure was used for measuring the performance. The main drivers of EO in SMEs were the personal work related values of the entrepreneur and his/her prior experience as an entrepreneur. The intrinsic work values related to interest, responsibility, challenge, self-development or intellectual stimulation and values related to status, power, achievement and recognition had a positive effect on the level of EO. On the other hand, extrinsic values related to high income, material possessions, benefits such as generous holidays, job security, and comfort through good working conditions decreased the level of EO

Drivers and barriers of mobile travel and tourism service adoption : a study of individual perceptions and business model development in a travel and tourism context

The Travel and Tourism field is undergoing changes due to the rapid development of information technology and digital services. Online travel has profoundly changed the way travel and tourism organizations interact with their customers. Mobile technology such as mobile services for pocket devices (e.g. mobile phones) has the potential to take this development even further. Nevertheless, many issues have been highlighted since the early days of mobile services development (e.g. the lack of relevance, ease of use of many services). However, the wide adoption of smartphones and the mobile Internet in many countries as well as the formation of so-called ecosystems between vendors of mobile technology indicate that many of these issues have been overcome. Also when looking at the numbers of downloaded applications related to travel in application stores like Google Play, it seems obvious that mobile travel and tourism services are adopted and used by many individuals. However, as business is expected to start booming in the mobile era, many issues have a tendency to be overlooked. Travelers are generally on the go and thus services that work effectively in mobile settings (e.g. during a trip) are essential. Hence, the individuals’ perceived drivers and barriers to use mobile travel and tourism services in on-site or during trip settings seem particularly valuable to understand; thus this is one primary aim of the thesis. We are, however, also interested in understanding different types of mobile travel service users. Individuals may indeed be very different in their propensity to adopt and use technology based innovations (services). Research is also switching more from investigating issues of mobile service development to understanding individuals’ usage patterns of mobile services. But designing new mobile services may be a complex matter from a service provider perspective. Hence, our secondary aim is to provide insights into drivers and barriers of mobile travel and tourism service development from a holistic business model perspective. To accomplish the research objectives seven different studies have been conducted over a time period from 2002 – 2013. The studies are founded on and contribute to theories within diffusion of innovations, technology acceptance, value creation, user experience and business model development. Several different research methods are utilized: surveys, field and laboratory experiments and action research. The findings suggest that a successful mobile travel and tourism service is a service which supports one or several mobile motives (needs) of individuals such as spontaneous needs, time-critical arrangements, efficiency ambitions, mobility related needs (location features) and entertainment needs. The service could be customized to support travelers’ style of traveling (e.g. organized travel or independent travel) and should be easy to use, especially easy to take into use (access, install and learn) during a trip, without causing security concerns and/or financial risks for the user. In fact, the findings suggest that the most prominent barrier to the use of mobile travel and tourism services during a trip is an individual’s perceived financial cost (entry costs and usage costs). It should, however, be noted that regulations are put in place in the EU regarding data roaming prices between European countries and national telecom operators are starting to see ‘international data subscriptions’ as a sales advantage (e.g. Finnish Sonera provides a data subscription in the Baltic and Nordic region at the same price as in Finland), which will enhance the adoption of mobile travel and tourism services also in international contexts. In order to speed up the adoption rate travel service providers could consider e.g. more local initiatives of free Wi-Fi networks, development of services that can be used, at least to some extent, in an offline mode (do not require costly network access during a trip) and cooperation with telecom operators (e.g. lower usage costs for travelers who use specific mobile services or travel with specific vendors). Furthermore, based on a developed framework for user experience of mobile trip arrangements, the results show that a well-designed mobile site and/or native application, which preferably supports integration with other mobile services, is a must for true mobile presence. In fact, travel service providers who want to build a relationship with their customers need to consider a downloadable native application, but in order to be found through the mobile channel and make contact with potential new customers, a mobile website should be available. Moreover, we have made a first attempt with cluster analysis to identify user categories of mobile services in a travel and tourism context. The following four categories were identified: info-seekers, checkers, bookers and all-rounders. For example “all-rounders”, represented primarily by individuals who use their pocket device for almost any of the investigated mobile travel services, constituted primarily of 23 to 50 year old males with high travel frequency and great online experience. The results also indicate that travel service providers will increasingly become multi-channel providers. To manage multiple online channels, closely integrated and hybrid online platforms for different devices, supporting all steps in a traveler process should be considered. It could be useful for travel service providers to focus more on developing browser-based mobile services (HTML5-solutions) than native applications that work only with specific operating systems and for specific devices. Based on an action research study and utilizing a holistic business model framework called STOF we found that HTML5 as an emerging platform, at least for now, has some limitations regarding the development of the user experience and monetizing the application. In fact, a native application store (e.g. Google Play) may be a key mediator in the adoption of mobile travel and tourism services both from a traveler and a service provider perspective. Moreover, it must be remembered that many device and mobile operating system developers want service providers to specifically create services for their platforms and see native applications as a strategic advantage to sell more devices of a certain kind. The mobile telecom industry has moved into a battle of ecosystems where device makers, developers of operating systems and service developers are to some extent forced to choose their development platforms.

The Influence of Network Competence on Internationalization of SMEs

This study focuses on the relationship between organizational network competence and the internationalization process of small- and medium sized enterprises (SMEs). Over recent decades, the global business environment has become increasingly conducive to internationalization of small firms. A central facilitating factor in the process has been the emergence of networked business relationships between internationalizing firms. Research on SME internationalization has found that certain types of structures and dynamics of business networks allow SMEs access to the resources they need to enter foreign markets. This consequently means that their internationalization often becomes to depend on the networks they are embedded in. However, research so far has mostly ignored the possibility that the organizational ability to develop and manage business network relationships, network competence, may be a major underlying factor in determining how well SMEs can leverage their network relationships to enter foreign markets and consequently may determine in large part how successful their internationalization process turns out to be. This study aims to respond to those gaps, by empirically examining how the development of network competence in internationalizing SMEs influences the internationalization outcomes that they can expect, and how such network competence is conceptualized and developed. Using a mixed methods approach, survey data collected from 298 Finnish SMEs across five industry sectors is first used to examine how levels of network competence are related to internationalization propensity of SMEs and their subsequent international performance, growth and profitability as internationally operating firms. In order to illustrate in more detail the ways in which network competence is conceptualized and how it develops during the internationalization process of an SME, qualitative data from internationally operating Finnish SMEs are used. Longitudinal interview data of an internationalizing Finnish SME is accompanied by data gathered through a series of semistructured interviews of Finnish and Russian managers involved in mutual business relationship dyads. Structurally, this thesis examines the research issue as an article-based dissertation, consisting of five journal and conference publications. Three of these publications are based on the quantitative data, and the remaining two apply the qualitative interview data. The results find several aspects where network competence has a positive influence on the success of internationalizing SMEs, how it develops and what it entails conceptually in this context. Quantitatively, the level of network competence is found to have a positive relationship to various internationalization outcomes, including the propensity of SMEs to enter foreign markets and on their subsequent international performance, their growth and their profitability. Additionally, the positive relationship is divided between the relationship-specific and cross-relational dimension of network competence, in that the influence of the former is relevant for the propensity to internationalize, while the latter is for the growth and profitability of the already internationalized SMEs. Qualitatively, the results suggest, firstly, that the development process of network competence does not necessarily precede the start of the internationalization process, but may occur through a gradual learning process alongside it. And secondly, the results also imply that the conceptualization of network competence by Finnish managers of internationally operating Finnish SMEs is structurally distinct from that of their culturally distinct partner managers in Russia. This study contributes to the literature on SME internationalization in several ways. Firstly, it introduces operationalized organizational competencies to the literature on internationalization of SMEs, which has so far mainly examined the influence of business networking on the internationalization process without having such an organizational viewpoint. Furthermore, this study provides a multi-level analysis of the determinants of successful SME internationalization, by examining various strategic and performance outcomes across the process. These results also contribute to the literature on organizational strategy of internationalizing SMEs, by clarifying how different dimensions of business networking may be optimal in different phases of the internationalization process. Conceptually, the results of this study contribute to the literature on competence development and SME internationalization, by illustrating how the development process of network competence may occur during internationalization process. Thus, they also contribute to the discussion on how SMEs are able to influence the dynamics and structures of their business networks over time. Finally, this study contributes to the literature on the role of culture in the internationalization process, by implying that the cultural background of the manager of the SME may determine whether business networking and network competence is seen as an organizational-level or an individual level capability. The study also includes some additional contributions to the literature on dynamic capabilities in strategic management, and on that of strategic business networks. These include further clarifying the exact nature and tangibility of dynamic capabilities, and being one of the first studies to introduce constructs from both dynamic capabilities and business network literature to the field of international entrepreneurship. And finally, the study also has some contribution on the two streams of literature, in illustrating how both dyadic and network-level capabilities may be relevant, depending on the current strategic goals and market position of the firm. Keywords: network competence, internationalizatio

The role of emotional state, sexual arousal, and alcohol intoxication as situational factors associated with adults’ online sexual contact with children and adolescents

In the present study we explored whether and how the situational factors; emotional states, sexual arousal, and alcohol intoxication influenced the propensity in adults to engage in online sexual contact with children (13 or younger) and adolescents (14 – 17 year olds). The results were compared to a group of adults that had engaged in online sexual contact with adults only (18 or older). We also looked at the variation over time within these situational factors during the online sexual contact with a child, an adolescent, or an adult. The present study was an online self-report survey to the adult populations in Finland, Sweden, and Germany, with a final sample (N = 776) of women and men who were active on the Internet. The participants were asked to report whether, how, and with whom they had engaged in online sexual contact. The results showed that more men than women reported online sexual contact with persons of all age groups, and that the situational factors; emotional states and sexual arousal influenced the propensity in both women and men to engage in online sexual contact with children and adolescents. However, the effects of alcohol intoxication were small and significant only for men. These results indicate that higher levels of emotional state and sexual arousal might increase the propensity to go against social norms and contact children and adolescents online for sexual purposes, but it can also imply that that those who look for online sexual contact online with children and adolescents, are more emotionally and/or sexually aroused than the group that only seek adult company or that these are post-hoc explanations for such sexual activities.

Periprocedural prognostic factors in coronary interventions – retrospective studies

Background: Approximately 11,000 revascularization procedures, either percutaneous coronary interventions (PCI) or coronary artery bypass grafting surgery (CABG), are performed yearly in Finland for coronary artery disease. Periprocedural risk factors for mortality and morbidity as well as long-term outcome have been extensively studied in general populations undergoing revascularization. Treatment choice between PCI and CABG in many high risk groups and risk-stratification, however, needs clarification and there is still room for improvement in periprocedural outcomes. Materials and methods: Cohorts of patients from Finnish hospitals revascularized between 2001 and 2011 were retrospectively analyzed. Patient records were reviewed for baseline variables and postprocedural outcomes (stroke, myocardial infarction, quality of life measured by the EQ-5D –questionnaire, repeat revascularization, bleeding episodes). Data on date and mode of death was acquired from Statistics Finland. Statistical analysis was performed to identify predictors of adverse events and compare procedures. Results: Postoperative administration of blood products (red blood cells, fresh frozen plasma, platelets) after isolated CABG independently and dose-dependently increases the risk of stroke. Patients 80 years or older who underwent CABG had better survival at 5 years compared to those who underwent PCI. After adjusting for baseline differences survival was similar. Patients on oral anticoagulation (OAC) for atrial fibrillation (AF) treated with CABG had better survival and overall outcome at 3 years compared to PCI patients. There was no difference in incidence of stroke or bleeding episodes. Differences in outcome remained significant after adjusting for propensity score. Lower health-related quality of life (HRQOL) scores as measured by the visual analogue scale (VAS) of the EQ-5D questionnaire at 6 months after CABG predicted later major adverse cardiac and cerebrovascular events (MACCE). Deteriorating function and VAS scores between 0 and 6 months on the EQ-5D also independently predicted later MACCE. Conclusions: Administration of blood products can increase the risk of stroke after CABG and liberal use of transfusions should be avoided. In the frail subpopulations of patients on OAC and octogenarians CABG appears to offer superior long-term outcome as compared to PCI. Deteriorating HRQOL scores predict later adverse events after CABG. Keywords: percutaneous coronary intervention, coronary artery bypass grafting, age over 80, transfusion, anticoagulants, coronary artery disease, health-related quality of life, outcome.

Adaptive Dynamics of Resource Specialization

Ecological specialization in resource utilization has various facades ranging from nutritional resources via host use of parasites or phytophagous insects to local adaptation in different habitats. Therefore, the evolution of specialization affects the evolution of most other traits, which makes it one of the core issues in the theory of evolution. Hence, the evolution of specialization has gained enormous amounts of research interest, starting already from Darwin’s Origin of species in 1859. Vast majority of the theoretical studies has, however, focused on the mathematically most simple case with well-mixed populations and equilibrium dynamics. This thesis explores the possibilities to extend the evolutionary analysis of resource usage to spatially heterogeneous metapopulation models and to models with non-equilibrium dynamics. These extensions are enabled by the recent advances in the field of adaptive dynamics, which allows for a mechanistic derivation of the invasion-fitness function based on the ecological dynamics. In the evolutionary analyses, special focus is set to the case with two substitutable renewable resources. In this case, the most striking questions are, whether a generalist species is able to coexist with the two specialist species, and can such trimorphic coexistence be attained through natural selection starting from a monomorphic population. This is shown possible both due to spatial heterogeneity and due to non-equilibrium dynamics. In addition, it is shown that chaotic dynamics may sometimes inflict evolutionary suicide or cyclic evolutionary dynamics. Moreover, the relations between various ecological parameters and evolutionary dynamics are investigated. Especially, the relation between specialization and dispersal propensity turns out to be counter-intuitively non-monotonous. This observation served as inspiration to the analysis of joint evolution of dispersal and specialization, which may provide the most natural explanation to the observed coexistence of specialist and generalist species.

Monooxygenase Diversity and Oxygen Activation within Polyketide Antibiotic Biosynthesis

Molecular oxygen (O2) is a key component in cellular respiration and aerobic life. Through the redox potential of O2, the amount of free energy available to organisms that utilize it is greatly increased. Yet, due to the nature of the O2 electron configuration, it is non-reactive to most organic molecules in the ground state. For O2 to react with most organic compounds it must be activated. By activating O2, oxygenases can catalyze reactions involving oxygen incorporation into organic compounds. The oxygen activation mechanisms employed by many oxygenases to have been studied, and they often include transition metals and selected organic compounds. Despite the diversity of mechanisms for O2 activation explored in this thesis, all of the monooxygenases studied in the experimental part activate O2 through a transient carbanion intermediate. One of these enzymes is the small cofactorless monooxygenase SnoaB. Cofactorless monooxygenases are unusual oxygenases that require neither transition metals nor cofactors to activate oxygen. Based on our biochemical characterization and the crystal structure of this enzyme, the mechanism most likely employed by SnoaB relies on a carbanion intermediate to activate oxygen, which is consistent with the proposed substrate-assisted mechanism for this family of enzymes. From the studies conducted on the two-component system AlnT and AlnH, both the functions of the NADH-dependent flavin reductase, AlnH, and the reduced flavin dependent monooxygenase, AlnT, were confirmed. The unusual regiochemistry proposed for AlnT was also confirmed on the basis of the structure of a reaction product. The mechanism of AlnT, as with other flavin-dependent monooxygenases, is likely to involve a caged radical pair consisting of a superoxide anion and a neutral flavin radical formed from an initial carbanion intermediate. In the studies concerning the engineering of the S-adenosyl-L-methionine (SAM) dependent 4-O-methylase DnrK and the homologous atypical 10-hydroxylase RdmB, our data suggest that an initial decarboxylation of the substrate is catalyzed by both of these enzymes, which results in the generation of a carbanion intermediate. This intermediate is not essential for the 4-O-methylation reaction, but it is important for the 10-hydroxylation reaction, since it enables substrate-assisted activation of molecular oxygen involving a single electron transfer to O2 from a carbanion intermediate. The only role for SAM in the hydroxylation reaction is likely to be stabilization of the carbanion through the positive charge of the cofactor. Based on the DnrK variant crystal structure and the characterizations of several DnrK variants, the insertion of a single amino acid in DnrK (S297) is sufficient for gaining a hydroxylation function, which is likely caused by carbanion stabilization through active site solvent restriction. Despite large differences in the three-dimensional structures of the oxygenases and the potential for multiple oxygen activation mechanisms, all the enzymes in my studies rely on carbanion intermediates to activate oxygen from either flavins or their substrates. This thesis provides interesting examples of divergent evolution and the prevalence of carbanion intermediates within polyketide biosynthesis. This mechanism appears to be recurrent in aromatic polyketide biosynthesis and may reflect the acidic nature of these compounds, propensity towards hydrogen bonding and their ability to delocalize π-electrons.