Dissertations / Theses on the topic 'Biochemical process'

Process algebra, formal specification, and model checking are all well studied techniques in the analysis of concurrent computer systems. More recently these techniques have been applied to the analysis of biochemical systems which, at an abstract level, have similar patterns of behaviour to concurrent processes. Process algebraic models and temporal logic specifications, along with their associated model-checking techniques, have been used to analyse biochemical systems. In this thesis we develop a spatio-temporal logic, the Logic of Behaviour in Context (LBC), for the analysis of biochemical models. That is, we define and study the application of a formal specification language which not only expresses temporal properties of biochemical models, but expresses spatial or contextual properties as well. The logic can be used to express, or specify, the behaviour of a model when it is placed into the context of another model. We also explore the types of properties which can be expressed in LBC, various algorithms for model checking LBC - each an improvement on the last, the implementation of the computational tools to support model checking LBC, and a case study on the analysis of models of post-translational biochemical oscillators using LBC. We show that a number of interesting and useful properties can be expressed in LBC and that it is possible to express highly useful properties of real models in the biochemistry domain, with practical application. Statements in LBC can be thought of as expressing computational experiments which can be performed automatically by means of the model checker. Indeed, many of these computational experiments can be higher-order meaning that one succinct and precise specification in LBC can represent a number of experiments which can be automatically executed by the model checker.

This thesis describes two main sections of work, an examination of a commercial product, Intrasite Gel, and the development of an algorithm for variable selection using projected latent structures.Following on from the successful development of a variable selection procedure for multivariate linear regression this work looks at transferring this idea for use with projected latent structures. The first part of this thesis will show how the variable selection algorithm was developed and used with three different data sets. The algorithm will be shown to be superior to standard projected latent structures, for linear multi-component data. Although the final algorithm developed requires considerable computing resources to carry out this is compensated for by significantly improved model predictions and robustness. The final algorithm developed is written to run using MATLAB on any computer platform that supports this application, though the principles of operation could be transferred to another method of execution, for example custom code written in C or Pascal. The approach used in the development of this method is that the ability of the model to predict unknownsamples is of far greater importance than the internal performance of the model. All the assessments of the procedures developed are based on the ability of the model to predict accurately and precisely samples that were not presented to the model during the training stage.The second section of this thesis is concerned with the study of Intrasite Gel, produced by Smith & Nephew Ltd. Hull. The material in question is a medical device intended to assist in the treatment and healing of wounds that are necrotic, sloughy or granulating. The product is characterised by its ability to maintain moisture equilibrium in a wound environment and to provide a suitable medium to encourage the growth of new cell tissue. Medical devices require registration, and as part of that registration a number of tests are made on samples to ensure that the material meets the required specifications. There was some concern at Smith & Nephew that the tests they were required to carry out as part of the device registration were not providing appropriate information about the product. Of particular interest was the fluid absorption property as it was suspected that the test has a large amount of random error associated with it and an investigation was required to examine this test and to provide an alternative procedure should the fluid absorption test prove inadequate. Also of interest to Smith & Nephew was the issue of sampling frequency, as it was felt that this should also be examined to determine whether the correct rate of sampling to ensure product quality was being carried out. The work reported here shows that the fluid absorption test as it stands is insufficient to the task of monitoring this property of Intrasite gel and that an alternative test should be considered. This work also showed that current sampling rate was too high and that the high sampling rate may in fact cause misleading assumptions as to the stability and quality of the product.

Automated tracking of cells across timelapse microscopy image sequences typically employs complex segmentation routines and/or bio-staining of the tracking objective. Often accurate identification of a cell's morphology is not of interest and the accurate segmentation of cells in pursuit of non-morphological parameters is complex and time consuming. This thesis explores the potential of internalized quantum dot nanoparticles as alternative, bio- and photo-stable optical markers for tracking the motions of cells through time. CdTe/ZnS core-shell quantum dots act as nodes in moving light display networks within A549, epithelial, lung cancer cells over a 40 hour time period. These quantum dot fluorescence sources are identified and interpreted using simplistic algorithms to find consistent, non-subjective centroids that represent cell centre locations. The presented tracking protocols yield an approximate 91% success rate over 24 hours and 78% over the full 40 hours. The nanoparticle moving light displays also provide simultaneous collection of cell motility data, resolution of mitotic traversal dynamics and identification of familial relationships enabling the construction of multi-parameter lineage trees. This principle is then developed further through inclusion of 3 different coloured quantum dots to create cell specific colour barcodes and reduce the number of time points necessary to successfully track cells through time. The tracking software and identification of parameters without detailed morphological knowledge is also demonstrated through automated extraction of DOX accumulation profiles and Cobalt agglomeration accruement statistics from two separate toxicology assays without the need for cell segmentation.

<p>In this master thesis, optical dissolved oxygen (DO) sensors are compared to standard electrochemical DO-sensors. The optical sensors are also compared against each other. DO is an important parameter to measure and to control during microorganism fermentation.  A DO-sensor needs to fulfill certain criteria, such as; purposive response time, the ability to show accurate and stable readings without drift over time, have the capability to manage several sterilizations and exhibiting membrane fouling resistance. The reference used to test these parameters was an industrially accepted electrochemical sensor. By comparing the two technologies, optical sensors showed similar abilities to perform regarding response time and drift performance during the longer test runs. But the optical sensors showed a better performance during sterilizations, with less drift in raw value. When using electrochemical sensors as a reference, different brands of optical sensor gave different results when testing for accuracy at different concentration levels. Due to error in the method, tests for fouling were inconclusive. However, membranes from the two kinds of technologies showed an equal hydrophobic character. The two participating optical DO sensors, Mettler Toledo InPro 6990i and Hamilton Visiferm were compared regarding ability to manage sterilizations and generate accurate measuring values at different DO concentrations. The two sensor brands showed an equal behavior during the fifteen test sterilizations. The Mettler Toledo InPro 6990 had better abilities to follow the linear behavior of the electrochemical reference sensor. While the  Hamilton Visiferm sensor differed at most 3.5% units from the electrochemical reference values. Hamilton Visiferm was tested with analogue signal, which gave a noise problem. When using digital signal the noise disappeared. The goal of this study was to recommend a sensor, for Belach Bioteknik AB, and the recommendation would be Hamilton Visiferm, for being practical as well as reasonable priced without renounce of measuring quality.</p>

[EN] This thesis is aimed to study the implications of the statistical modeling approaches proposed for the bilinear modeling of batch processes, develop new techniques to overcome some of the problems that have not been yet solved and apply them to data of biochemical processes. The study, discussion and development of the new methods revolve around the four steps of the modeling cycle, from the alignment, preprocessing and calibration of batch data to the monitoring of batches trajectories. Special attention is given to the problem of the batch synchronization, and its effect on the modeling from different angles. The manuscript has been divided into four blocks. First, a state-of- the-art of the latent structures based-models in continuous and batch processes and traditional univariate and multivariate statistical process control systems is carried out. The second block of the thesis is devoted to the preprocessing of batch data, in particular, to the equalization and synchronization of batch trajectories. The first section addresses the problem of the lack of equalization in the variable trajectories. The different types of unequalization scenarios that practitioners might finnd in batch processes are discussed and the solutions to equalize batch data are introduced. In the second section, a theoretical study of the nature of batch processes and of the synchronization of batch trajectories as a prior step to bilinear modeling is carried out. The topics under discussion are i) whether the same synchronization approach must be applied to batch data in presence of different types of asynchronisms, and ii) whether synchronization is always required even though the length of the variable trajectories are constant across batches. To answer these questions, a thorough study of the most common types of asynchronisms that may be found in batch data is done. Furthermore, two new synchronization techniques are proposed to solve the current problems in post-batch and real-time synchronization. To improve fault detection and classification, new unsupervised control charts and supervised fault classifiers based on the information generated by the batch synchronization are also proposed. In the third block of the manuscript, a research work is performed on the parameter stability associated with the most used synchronization methods and principal component analysis (PCA)-based Batch Multivariate Statistical Process Control methods. The results of this study have revealed that accuracy in batch synchronization has a profound impact on the PCA model parameters stability. Also, the parameter stability is closely related to the type of preprocessing performed in batch data, and the type of model and unfolding used to transform the three-way data structure to two-way. The setting of the parameter stability, the source of variability remaining after preprocessing and the process dynamics should be balanced in such a way that multivariate statistical models are accurate in fault detection and diagnosis and/or in online prediction. Finally, the fourth block introduces a graphical user-friendly interface developed in Matlab code for batch process understanding and monitoring. To perform multivariate analysis, the last developments in process chemometrics, including the methods proposed in this thesis, are implemented.<br>[ES] La presente tesis doctoral tiene como objetivo estudiar las implicaciones de los métodos estadísticos propuestos para la modelización bilineal de procesos por lotes, el desarrollo de nuevas técnicas para solucionar algunos de los problemas más complejos aún por resolver en esta línea de investigación y aplicar los nuevos métodos a datos provenientes de procesos bioquímicos para su evaluación estadística. El estudio, la discusión y el desarrollo de los nuevos métodos giran en torno a las cuatro fases del ciclo de modelización: desde la sincronización, ecualización, preprocesamiento y calibración de los datos, a la monitorización de las trayectorias de las variables del proceso. Se presta especial atención al problema de la sincronización y su efecto en la modelización estadística desde distintas perspectivas. El manuscrito se ha dividido en cuatro grandes bloques. En primer lugar, se realiza una revisión bibliográfica de las técnicas de proyección sobre estructuras latentes para su aplicación en procesos continuos y por lotes, y del diseño de sistemas de control basados en modelos estadísticos multivariantes. El segundo bloque del documento versa sobre el preprocesamiento de los datos, en concreto, sobre la ecualización y la sincronización. La primera parte aborda el problema de la falta de ecualización en las trayectorias de las variables. Se discuten las diferentes políticas de muestreo que se pueden encontrar en procesos por lotes y las soluciones para ecualizar las variables. En la segunda parte de esta sección, se realiza un estudio teórico sobre la naturaleza de los procesos por lotes y de la sincronización de las trayectorias como paso previo a la modelización bilineal. Los temas bajo discusión son: i) si se debe utilizar el mismo enfoque de sincronización en lotes afectados por diferentes tipos de asincronismos, y ii) si la sincronización es siempre necesaria aún y cuando las trayectorias de las variables tienen la misma duración en todos los lotes. Para responder a estas preguntas, se lleva a cabo un estudio exhaustivo de los tipos más comunes de asincronismos que se pueden encontrar en este tipo de datos. Además, se proponen dos nuevas técnicas de sincronización para resolver los problemas existentes en aplicaciones post-morten y en tiempo real. Para mejorar la detección de fallos y la clasificación, también se proponen nuevos gráficos de control no supervisados y clasificadores de fallos supervisados en base a la información generada por la sincronización de los lotes. En el tercer bloque del manuscrito se realiza un estudio de la estabilidad de los parámetros asociados a los métodos de sincronización y a los métodos estadístico multivariante basados en el Análisis de Componentes Principales (PCA) más utilizados para el control de procesos. Los resultados de este estudio revelan que la precisión de la sincronización de las trayectorias tiene un impacto significativo en la estabilidad de los parámetros de los modelos PCA. Además, la estabilidad paramétrica está estrechamente relacionada con el tipo de preprocesamiento realizado en los datos de los lotes, el tipo de modelo a justado y el despliegue utilizado para transformar la estructura de datos de tres a dos dimensiones. El ajuste de la estabilidad de los parámetros, la fuente de variabilidad que queda después del preprocesamiento de los datos y la captura de las dinámicas del proceso deben ser a justados de forma equilibrada de tal manera que los modelos estadísticos multivariantes sean precisos en la detección y diagnóstico de fallos y/o en la predicción en tiempo real. Por último, el cuarto bloque del documento describe una interfaz gráfica de usuario que se ha desarrollado en código Matlab para la comprensión y la supervisión de procesos por lotes. Para llevar a cabo los análisis multivariantes, se han implementado los últimos desarrollos en la quimiometría de proc<br>[CAT] Aquesta tesi doctoral te com a objectiu estudiar les implicacions dels mètodes de modelització estadística proposats per a la modelització bilineal de processos per lots, el desenvolupament de noves tècniques per resoldre els problemes encara no resolts en aquesta línia de recerca i aplicar els nous mètodes a les dades dels processos bioquímics. L'estudi, la discussió i el desenvolupament dels nous mètodes giren entorn a les quatre fases del cicle de modelització, des de l'alineació, preprocessament i el calibratge de les dades provinents de lots, a la monitorització de les trajectòries. Es presta especial atenció al problema de la sincronització per lots, i el seu efecte sobre el modelatge des de diferents angles. El manuscrit s'ha dividit en quatre grans blocs. En primer lloc, es realitza una revisió bibliogràfica dels principals mètodes basats en tècniques de projecció sobre estructures latents en processos continus i per lots, així com dels sistemes de control estadístics multivariats. El segon bloc del document es dedica a la preprocessament de les dades provinents de lots, en particular, l' equalització i la sincronització. La primera part aborda el problema de la manca d'equalització en les trajectòries de les variables. Es discuteixen els diferents tipus d'escenaris en que les variables estan mesurades a distints intervals i les solucions per equalitzar-les en processos per lots. A la segona part d'aquesta secció es porta a terme un estudi teòric de la naturalesa dels processos per lots i de la sincronització de les trajectòries de lots com a pas previ al modelatge bilineal. Els temes en discussió són: i) si el mateix enfocament de sincronització ha de ser aplicat a les dades del lot en presència de diferents tipus de asincronismes, i ii) si la sincronització sempre es requereix tot i que la longitud de les trajectòries de les variables són constants en tots el lots. Per respondre a aquestes preguntes, es du a terme un estudi exhaustiu dels tipus més comuns de asincronismes que es poden trobar en les dades provinents de lots. A més, es proposen dues noves tècniques de sincronització per resoldre els problemes existents la sincronització post-morten i en temps real. Per millorar la detecció i la classificació de anomalies, també es proposen nous gràfics de control no supervisats i classificadors de falla supervisats dissenyats en base a la informació generada per la sincronització de lots. En el tercer bloc del manuscrit es realitza un treball de recerca sobre l'estabilitat dels paràmetres associats als mètodes de sincronització i als mètodes estadístics multivariats basats en l'Anàlisi de Components Principals (PCA) més utilitzats per al control de processos. Els resultats d'aquest estudi revelen que la precisió en la sincronització per lots te un profund impacte en l'estabilitat dels paràmetres dels models PCA. A més, l'estabilitat paramètrica està estretament relacionat amb el tipus de preprocessament realitzat en les dades provinents de lots, el tipus de model i el desplegament utilitzat per transformar l'estructura de dades de tres a dos dimensions. L'ajust de l'estabilitat dels paràmetres, la font de variabilitat que queda després del preprocessament i la captura de la dinàmica de procés ha de ser equilibrada de tal manera que els models estadístics multivariats són precisos en la detecció i diagnòstic de fallades i/o en la predicció en línia. Finalment, el quart bloc del document introdueix una interfície gràfica d'usuari que s'ha dissenyat e implementat en Matlab per a la comprensió i la supervisió de processos per lots. Per dur a terme aquestes anàlisis multivariats, s'han implementat els últims desenvolupaments en la quimiometria de processos, incloent-hi els mètodes proposats en aquesta tesi.<br>González Martínez, JM. (2015). ADVANCES ON BILINEAR MODELING OF BIOCHEMICAL BATCH PROCESSES [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/55684<br>TESIS<br>Premiado

The cell is a highly ordered and intricate machine within which a wide variety of chemical processes take place. The full scientific understanding of cellular physiology requires accurate mathematical models that depict the temporal dynamics of these chemical processes. Modelers build mathematical models of chemical processes primarily from systems of differential equations. Although developing new biological ideas is more of an art than a science, constructing a mathematical model from a biological idea is largely mechanical and automatable. This dissertation describes the practices and processes that biological modelers use for modeling and simulation. Computational biologists struggle with existing tools for creating models of complex eukaryotic cells. This dissertation develops new processes for biological modeling that make model creation, verification, validation, and testing less of a struggle. This dissertation introduces computational software that automates parts of the biological modeling process, including model building, transformation, execution, analysis, and evaluation. User and methodological requirements heavily affect the suitability of software for biological modeling. This dissertation examines the modeling software in terms of these requirements. Intelligent, automated model evaluation shows a tremendous potential to enable the rapid, repeatable, and cost-effective development of accurate models. This dissertation presents a case study that indicates that automated model evaluation can reduce the evaluation time for a budding yeast model from several hours to a few seconds, representing a more than 1000-fold improvement. Although constructing an automated model evaluation procedure requires considerable domain expertise and skill in modeling and simulation, applying an existing automated model evaluation procedure does not. With this automated model evaluation procedure, the computer can then search for and potentially discover models superior to those that the biological modelers developed previously.<br>Ph. D.

The cell is a highly ordered and intricate machine within which a wide variety of chemical processes take place. The full scientific understanding of cellular physiology requires accurate mathematical models that depict the temporal dynamics of these chemical processes. Modelers build mathematical models of chemical processes primarily from systems of differential equations. Although developing new biological ideas is more of an art than a science, constructing a mathematical model from a biological idea is largely mechanical and automatable. This dissertation describes the practices and processes that biological modelers use for modeling and simulation. Computational biologists struggle with existing tools for creating models of complex eukaryotic cells. This dissertation develops new processes for biological modeling that make model creation, verification, validation, and testing less of a struggle. This dissertation introduces computational software that automates parts of the biological modeling process, including model building, transformation, execution, analysis, and evaluation. User and methodological requirements heavily affect the suitability of software for biological modeling. This dissertation examines the modeling software in terms of these requirements. Intelligent, automated model evaluation shows a tremendous potential to enable the rapid, repeatable, and cost-effective development of accurate models. This dissertation presents a case study that indicates that automated model evaluation can reduce the evaluation time for a budding yeast model from several hours to a few seconds, representing a more than 1000-fold improvement. Although constructing an automated model evaluation procedure requires considerable domain expertise and skill in modeling and simulation, applying an existing automated model evaluation procedure does not. With this automated model evaluation procedure, the computer can then search for and potentially discover models superior to those that the biological modelers developed previously.<br>Ph. D.

Ammonia (NH3) in air is of major health and environmental concern, i.e. contribution to the greenhouse effect when NH3 is converted to nitrous oxide (N2O) in the atmosphere. Another important concern for ammonia in the atmosphere, is the possible conversion to secondary fine particulate matter in the presence of SOx or NOx. The main sources of NH3 in air are waste and food processing industries as well as animal livestock production. The increase in environmental litigation, coupled with rising environmental awareness and a focus on quality of life, has resulted in an increase in the investigation and implementation of new technologies for the treatment of ammonia. In addition, existing methods are cost intensive, unreliable or complex and difficult to control. In this research, a novel biochemical ammonia removing process has been developed and operated for 300 days. This process involves a sequence of biological, physical and chemical processes initiated by the dissolution of the introduced ammonia into the water of the filter system to make the ammonium available for biological degradation (nitrification). A spontaneous reaction of the intermediate nitrification product, nitrite, and ammonium to nitrogen takes place as soon as both compounds are present. This reaction is known to be catalysed by zeolite (clinoptilolite). Water from the discharged moisture of the filter system was condensed on top of the reactor by employing a novel moisture control mechanism. The clean condensate percolated by gravity through the reactor bed and forced the accumulated compounds to the bottom of the reactor. This led to a gradient distribution of compounds across the reactor depth with the highest concentrations at the bottom (140 mM ammonium, 1 M nitrite and 350 mM nitrate), favouring the chemical reaction of NH4+ + NO2-

Thesis (PhD (Process Engineering))--University of Stellenbosch, 2010.<br>ENGLISH ABSTRACT: Biological agents offer several opportunities to address the many challenges faced in intensive reticulated aquaculture. We therefore isolated and selected Bacillus spp. as potential biological agents, because this group has demonstrated an array of biological activities of possible benefit to aquaculture. They also display advantages in terms of robustness during bioprocessing and end product application. Natural isolates obtained from Cyprinus carpio, selected as a model high-value ornamental fish species, and associated culture environments, were purified and assessed for in vitro efficacy based on the inhibition of growth of pathogenic Aeromonas hydrophila and the decrease in concentrations of ammonium, nitrite, nitrate and phosphate ions, typically found as waste products in aquaculture systems. Based on suitability for aquaculture application, isolates B001, B002 and B003 were selected and further evaluated in vitro and in an in vivo trial with C. carpio. Inhibition of Aer. hydrophila growth and a decrease in concentrations of waste ions were demonstrated in these studies. Based on 16S RNA sequence homology, the isolates were identified as Bacillus subtilis, B. cereus and B. licheniformis, respectively. High sequence homology between B. subtilis and B. anthracis necessitated further safety studies on the best isolate, B. cereus NRRL100132 (B002). The isolate was shown not to contain the anthrax virulence genes pOX1, pOX2 or the B. cereus enterotoxin. Elucidation of the potential modes of action of a biological agent facilitates an understanding of functionality and encourages technology uptake by end users. Competitive exclusion through growth rate and competitive uptake of glucose and iron, the latter facilitated by siderophore production, were shown to be key mechanisms at play in inhibition of Aer. hydrophila by the B. cereus isolate. As production cost is an important consideration in development of commercially relevant biological products, we examined the optimization of nutrient supplementation, which has an impact on high-density production of spores by fermentation. Corn steep liquor (CSL) was identified as a lower cost and more effective nutrient source in comparison to conventional nutrient substrates, in particular yeast extract and nutrient broth. The improved sporulation performance of B. cereus could be related to the increased availability of free amino acids, carbohydrates, and minerals in CSL, which had a positive effect on organism growth and sporulation efficiency. The impact of nutrient concentration on spore yield and productivity was modelled to develop a tool for selection of optimal conditions. Excellent correlation with actual laboratory fermentation data was demonstrated. A cost analysis revealed that production using liquid phytase treated and ultra-filtered CSL was less expensive than spray dried CSL and supported cultivation of B. cereus spores at densities higher than 1×1010 CFU ml 1. Adoption of biological agents in commercial applications is lacking, due to limitations in process and product development that address key end user product requirements such as cost, efficacy, shelf life and convenience. The development of suitable spore recovery, drying, formulation and tablet production process steps was thus performed. Key criteria used for downstream process unit evaluation included spore viability, recovery, spore balance closure, spore re-germination, product intermediate stability, end product stability and efficacy. A process flow sheet comprising vertical tube centrifugation, fluidised bed agglomeration and tablet pressing yielded an attractive product. The formulation included corn steep liquor and glucose to enhance subsequent spore re-germination. Viable spore recovery and spore balance closure across each of the process units was high (>70% and >99% respectively), with improvement in recovery possible by adoption of continuous processing at large scale. Spore re-germination was 97%, whilst a product half-life in excess of 5 years was estimated based on thermal resistance curves. The process resulted in a commercially attractive product and affordable variable cost of production. Functionality of the product, incorporating the B. cereus isolate, was investigated across a range of physiological conditions, including salinity, pH and temperature, based on rearing of C. carpio. Temperature had a significant influence on germination, specific growth rate and increase in cell number of B. cereus, whilst salinity and pH did not have any measurable effect on growth. Controlled studies in bioreactors and modelling of the data to the Arrhenius function indicated the existence of high and low growth temperature domains. The rates of pathogenic Aer. hydrophila suppression and decrease in waste ion concentrations (ammonium, nitrite, nitrate and phosphate) were translated into a linear predictive indicator of efficacy of the B. cereus isolate at different temperatures. This study has resulted in development of an upstream and downstream process for production of a new B. cereus isolate (NRRL 100132) which was shown to be safe, stable, functional, robust and cost effective for application in aquaculture.<br>AFRIKAANSE OPSOMMING: Biologiese middels bied verskeie maniere om die veelvoudige uitdagings van intensiewe netsgewyse akwakultuur aan te spreek. Gevolglik het ons uitgesoekte Bacillus spesies as potensiële biologiese middels geïsoleer, omdat hierdie groep verskeie biologiese aktiwiteite demonstreer wat van potensiële waarde kan wees in akwakultuur. Die groep toon ook voordele in terme van robuustheid gedurende bioprosessering en eind-toepassings. Natuurlike bakteriële isolate vanuit Cyprinus carpio geassosieerde kultuur omgewings, geselekteer as 'n hoë-waarde model ornamentele spesie, is gesuiwer. Die in vitro doeltreffendheid van die isolate is bepaal gebasseerd op die groei inhibisie van patogeniese Aeromons hydrophila asook die afname in konsentrasies van ammonium, nitriete, nitrate en fosfaat ione wat as tipiese afval produkte gevind word in akwakultuur sisteme. Isolate B001, B002 en B003 is geselekteer op grond van geskiktheid en verder evalueer in in vitro en in vivo proewe met C. carpio. Groei inhibisie van Aer. hydrophila asook 'n afname in konsentrasies van afval ione was tydens die studies gedemonstreer is. Die isolate is identifiseer as Bacillus subtilis, B. cereus en B. licheniformis, respektiewelik, op grond van 16S RNS volgorde homologie. Die hoë volgorde homologie tussen B. subtilis en B. anthracis het verdere veiligheidstudies op die beste isolaat, B. cereus NRRL100132 (B002) genoodsaak. Die isolaat het nie die antraks virulensie plasmied pOX1, pOX2 of die B. cereus enterotoksien getoon nie. Uitklaring van die potensiële meganismes van aksie van biologiese middels fasiliteer 'n begrip van funksionaliteit en moedig tegnologie aanvaarding deur eind-gebruikers aan. Mededingende uitsluiting deur groeitempo en mededingende opname van glukose asook die produksie van siderofore is bewys as sleutel meganismes betrokke in die inhibisie van Aer. hydrophila deur die B. cereus isolaat. Aangesien koste 'n belangrike oorweging is in die ontwikkeling van kommersiële toepaslike biologiese produkte, is die optimisering van voedingstof aanvullings wat 'n impak het op hoëdigtheid produksie van spore deur fermentasie ondersoek. Week-vloeistof van mielie prosessering (CSL) is identifiseer as 'n lae koste en effektiewe voedingsbron in vergelyking met konvensionele voeding substrate, veral gisekstrak en voedingsboeljon. Die verbeterde sporulering prestasie van B. cereus kon toegeskryf word aan die verhoogde beskikbaarheid van vrye aminosure, koolhidrate en minerale in CSL, wat 'n postitewe effek op organisme groei en sporulerings effektiwiteit getoon het. Die impak van voedingstof konsentrasie op spoor opbrengs en produktiwiteit is gemodelleer om 'n werktuig vir die selektering van optimale kondisies te ontwikkel. Uitstekende korrelasie met werklike laboratorium data is gedemonstreer. Koste analises het getoon dat produksie deur middel van vloeibare fitase-behandelde en ultra-filtreerde CSL goedkoper is as sproei-gedroogde CSL en ondersteun verder die kultivering van B. cereus spore teen digthede hoër as 1 x 1010 kolonie vormende eenhede.ml-1. Die opname van biologiese middels in kommersiële toepassings skiet tekort as gevolg van beperkinge in proses en produk ontwikkeling wat belangrike eind-gebruiker vereistes soos koste, doeltreffendheid, rak leeftyd en gerieflikheid aanspreek. Die ontwikkeling van toepaslike prosesse vir spoor herwinning, droging, formulering en tablet produksie is gevolglik uitgevoer. Belangrike maatstawwe wat gebruik is vir stroomaf proseseenheid-ontwikkeling het lewensvatbaarheid, herwinning, spoor balans sluiting, spoor her-ontkieming, intermediêre produk stabiliteit, eindproduk stabiliteit en doeltreffendheid ingesluit. 'n Proses vloeidiagram bestaande uit vertikale buis sentrifugasie, vloeibare bed agglomerasie en tablet persing het 'n aantreklike produk voortgebring. Die formulering het ook CSL en glukose ingesluit om gevolglike spoor herontkieming te verbeter. Lewensvatbare spoor herwinning en spoor balans sluiting oor elke proses eenheid was hoog (>70% en 99% respektiewelik) met verbetering in herwinning wat moontlik gemaak is deur die gebruik van aaneenlopende prosessering op groot skaal. Spoor her-ontkieming was 97%, terwyl produk halfleeftyd langer as 5 jaar beraam is, gebasseer op termiese weerstand grafieke. Die proses het gelei tot 'n kommersiële aantreklike produk asook bekostigbare veranderbare produksie koste. Die funksionaliteit van die tablet-produk met die ingeslote B. cereus isolaat is ondersoek oor 'n reeks fisiologiese kondisies insluitend soutgehalte, pH en temperatuur, gebasseer op die kultivering van C. carpio. Temperatuur het 'n betreklike invloed op ontkieming, spesifieke groeitempo en toename in sel hoeveelheid van B. cereus gehad, terwyl soutgehalte en pH nie enige meetbare effek op groei gehad het nie. Gekontrolleerde studies in bioreaktors en modellering van die data op die Arrhenius funksie het hoë en lae groei temperatuur domeins gewys. Die tempo van patogeniese Aer. hydrophila onderdrukking en afname in konsentrasies van afval-ione (ammonium, nitriete, nitrate en fosfaat) is herlei na 'n liniêre voorspellende aanwysing van effektiwiteit van B. cereus isolate by verskillende temperature. Die studie het gelei tot die ontwikkeling van stroomop- en stroomaf-prosesse vir die produksie van 'n nuwe B. cereus isolaat (NRRL 100132) wat bewys is as veilig, stabiel, funksioneel, robuust en koste effektiewe vir toepassing in akwakultuur.

<p>The goals of this Bachelors’ Thesis were to establish the production loss through the VTIS1 Pasteur at Arla Foods Linköping dairy and then suggest simple but effective solutions to minimize the losses for each production run. If the timeframe allowed it, some of the solutions would be implemented and tested in full production to determine their impact on economic and environmental savings.</p><p>The very first step of this project was to establish the statistics over what losses there is in the production pipeline through VTIS1, from milk and powders in mixer tanks to finished and packed products in cold storage. Then, when knowing how much product was actually lost and where, those bottlenecks could be pinpointed and examined more closely. At this stage the focus was switched, from the otherwise complicated and broad manufacturing process, to the VTIS1-aggregate, since early data indicated there were quite substantial losses there.</p><p>In conjunction with this project Arla Foods purchased an absorption photometer-measuring device (OPTEK AS16) that could be inserted straight into the product stream out of VTIS1, allowing real time measurements of its optical density. A substantial series of measurements of the product stream (through all of November) indicated that a certain amount of pure quality product was discarded into the factory sewers with each production batch. Volumes of the product losses clearly depended on which sterile tank it was sent to, due to the different lengths of the pipelines.</p><p>Through the production year of 2007 the VTIS1 aggregate pasteurised over 1500 batches of various products and all preliminary data indicates this number to increase significantly in 2008. Since most products going through VTIS1 contain both milk/cream and some flavourings or jams in various compositions, this means they are a fair bit more expensive per litre in comparison to plain milk.</p><p>Since the VTIS1 aggregate is positioned fairly early in the production line it was deemed logical to start there to eliminate losses and then, if time would allow, move further down the stream.</p><p>The time assigned for the project did allow for both creating a statistics database over product losses and the implementations of a few suggested adjustments. The report includes the estimated financial savings of this project for 2008; the numbers are based on how much the adjustments would have saved Arla Foods in economic losses through 2007. This includes the costs for the pure volumes of products as well as the estimated savings of environmental costs for waste treatment during that same period, based on the products’ COD, S-tot, N-tot and Ts contents.</p><br><p>Examensarbetets syfte har bestått i att med statistik kartlägga produktförlusterna i genom Arla Foods i Linköpings VTIS1 produktion och föreslå möjliga lösningar för att minska råvarusvinnet. Detta skall åstadkommas med hjälp av nya smarta lösningar på processoptimeringar, ändringar i körrutiner och bättre produktplanering.</p><p>Projektbeskrivningen var ursprungligen med avsikt ganska vag, för att sedan efter litet efterforskning fokusera uppmärksamheten på området kring Mixertankarna, VTIS1 och Steriltankarna. I studien har vikten lagts på rent tekniska svinn, som sker vid varje körning och inte mätbart påverkas av den mänskliga faktorn, detta för att få en repeterbarhet och stabila mätvärden.</p><p>Statistiksammanställning och kartering av de olika produktförlusterna gjordes i Excel, där data hämtades både via Arla Foods AS400 server samt logisticprogrammet Movex Explorer som hanterar lagerförningar, men även via ABB: s WebProInfo server och trendverktyget MES.</p><p>En snabb analys av den förenklade statistiken visade på att mycket av de förluster som uppstod i tillverkningen genom VTIS1 faktiskt uppstod i själva aggregatet och det var alltså här man beslutade att, i samråd med Timo Kjellberg och Gunnar Jangbrand, begränsa projektet till just mixertankarna, VTIS och steriltankarna.</p><p>Vid planeringen av examensarbetet var tanken att gå igenom hela tillverkningsprocessen, från det att produkten blandats och behandlats, till det att den förpackats och transporterats till lagerlokalen.</p><p>Av tidstekniska skäl har alltså tidfördelningen fokuserats till att omfatta en liten men dock betydelsefull del i produktionen. Processen i sin helhet är alltför komplex för att sätta sig in i på såpass kort tid och detta tillvägagångssätt har gett goda resultat, mycket på grund av att den tid som funnits kunnat fokuseras på att noga dokumentera, analysera och förbättra tillverkningsprocessen från första början.</p><p>Målsättningen för arbetet är att kunna minimera det tekniska svinn som uppkommer i varje ny produktionscykel och på så sätt både spara Arlas pengar och miljön, en faktor som i rapporten har räknats om till en ren ekonomisk besparing i form av minskade miljöavgifter. Fler möjliga förbättringar tas upp senare i rapporten.</p>

Membranous tubules are very abundant structures in living cells and form or are part of most intracellular organelles. The Golgi apparatus is mainly formed by tubules, which adopt different geometries and conformations. However, their physiological role has not yet been established and this is mainly due to the almost absolute lack of knowledge about the biochemical mechanisms regulating their formation, maintenance and disruption. The aim of this thesis was to investigate in a systematic way these mechanisms. The first step has been to set up an in vitro morphological assay suitable for the visualisation of Golgi-associated tubules in isolated Golgi stacks. This assay was based on electron microscopy and specifically on negative staining of whole-mount preparations. It allowed both qualitative and quantitative analysis of the morphological changes of Golgiassociated tubules after in vitro incubations. This assay was then used for screening several molecules or experimental conditions for their effect on tubular homeostasis. Among them, the most significant was BARS (BFA-dependent ADP-Ribosylation Substrate), a protein previously implicated in the maintenance of Golgi architecture. BARS has been found to cause the selective breakdown of the tubular part of the Golgi complex promoting fission events which convert the tubular structures into clusters of vesicles. This effect correlated with the enzymatic activity of BARS, which acts as an acyl-CoA dependent lysophosphatidic acid acyl transferase (LPAAT), increasing phosphatidic acid (PA) levels in Golgi membranes. This suggests that local modifications of the composition of the lipid bilayer is a possible mechanism for the fission of membranous tubules.

Thesis (MTech (Chemical Engineering))--Cape Peninsula University of Technology, 2016.<br>Grape producers and wine makers in South Africa are currently affected by various challenges, which include anti-alcohol lobbies, climate change, over-production in some vintages and the lack of transformation including empowerment in certain sectors of the industry. Climate change and global warming lead to poor quality wine grapes and as a result, poor quality wine. Therefore, there is a need to channel grapes away from normal wine production and provide an alternative source of income for the industry. The overall aim of this study was therefore to provide an alternative outlet for overproduced wine grapes by producing balsamic-styled vinegar (BSV) in South Africa. Balsamic vinegar is different from other vinegars because it is a direct product of grape must and not a downstream or by-product of wine production. Balsamic vinegar entails lower production costs when compared to the production of wine due to the low technological process requirements during production; therefore, this could be an opportunity for small business entrepreneurs with low capital start-up. In addition, balsamic vinegar can command a high price, which is a benefit for grape producers. The primary aim of this investigation was to biochemically analyse a BSV production process in which 5 non-Saccharomyces yeast and 15 acetic acid bacteria (AAB) were used for a multicultural alcoholic-acetous (EtOH-AcOH) fermentation process. To achieve this aim, a fermentation process was designed where the data generated was fitted into kinetic models and the proliferation including the population dynamics of the microbial consortia were studied.

Urease is a nickel-dependent enzyme that catalyzes hydrolysis of urea in the last step of organic nitrogen mineralization. Its active site contains a dinuclear center for Ni(II) ions that must be inserted into the apo-enzyme through the action of four accessory proteins (UreD, UreE, UreF, UreG) leading to activation of urease. UreE, acting as a metallo-chaperone, delivers Ni(II) to the preformed complex of apo-urease-UreDFG and has the capability to enhance the GTPase activity of UreG. This study, focused on characterization of UreE from Sporosarcina pasteurii (SpUreE), represents a piece of information on the structure/mobility-function relationships that control nickel binding by SpUreE and its interaction with SpUreG. A calorimetric analysis revealed the occurrence of a binding event between these proteins with positive cooperativity and a stoichiometry consistent with the formation of the (UreE)2-(UreG)2 hetero-oligomer complex. Chemical Shift Perturbations induced by the protein-protein interaction were analyzed using high-resolution NMR spectroscopy, which allowed to characterize the molecular details of the protein surface of SpUreE involved in the complex formation with SpUreG. Moreover, backbone dynamic properties of SpUreE, determined using 15N relaxation analysis, revealed a general mobility in the nanoseconds time-scale, with the fastest motions observed at the C-termini. The latter analysis made it possible for the first time to characterize of the C-terminal portions, known to contain key residues for metal ion binding, that were not observed in the crystal structure of UreE because of disorder. The residues belonging to this portion of SpUreE feature large CSPs upon addition of SpUreG, showing that their chemical environment is directly affected by protein-protein interaction. Metal ion selectivity and affinity of SpUreE for cognate Ni(II) and non cognate Zn(II) metal ions were determined, and the ability of the protein to select Ni(II) over Zn(II), in consistency with the proposed role in Ni(II) cations transport, was established.

Urease is a nickel-dependent enzyme that catalyzes hydrolysis of urea in the last step of organic nitrogen mineralization. Its active site contains a dinuclear center for Ni(II) ions that must be inserted into the apo-enzyme through the action of four accessory proteins (UreD, UreE, UreF, UreG) leading to activation of urease. UreE, acting as a metallo-chaperone, delivers Ni(II) to the preformed complex of apo-urease-UreDFG and has the capability to enhance the GTPase activity of UreG. This study, focused on characterization of UreE from Sporosarcina pasteurii (SpUreE), represents a piece of information on the structure/mobility-function relationships that control nickel binding by SpUreE and its interaction with SpUreG. A calorimetric analysis revealed the occurrence of a binding event between these proteins with positive cooperativity and a stoichiometry consistent with the formation of the (UreE)2-(UreG)2 hetero-oligomer complex. Chemical Shift Perturbations induced by the protein-protein interaction were analyzed using high-resolution NMR spectroscopy, which allowed to characterize the molecular details of the protein surface of SpUreE involved in the complex formation with SpUreG. Moreover, backbone dynamic properties of SpUreE, determined using 15N relaxation analysis, revealed a general mobility in the nanoseconds time-scale, with the fastest motions observed at the C-termini. The latter analysis made it possible for the first time to characterize of the C-terminal portions, known to contain key residues for metal ion binding, that were not observed in the crystal structure of UreE because of disorder. The residues belonging to this portion of SpUreE feature large CSPs upon addition of SpUreG, showing that their chemical environment is directly affected by protein-protein interaction. Metal ion selectivity and affinity of SpUreE for cognate Ni(II) and non cognate Zn(II) metal ions were determined, and the ability of the protein to select Ni(II) over Zn(II), in consistency with the proposed role in Ni(II) cations transport, was established.

A scientifically sound method for calculating the parameters of bio-cleaning should contain as a basic a reliable mathematical description of the stationary process. The results of stationary laboratory experiments are presented in the coordinates “specific rate of destruction V – concentration ρ”. Statistical processing proves the presence of substrate inhibition for both gaseous and soluble and dissolved harmful substances in water. For an analytical description of the dependence of the biooxidation rate on the concentration of contaminants, a phenomenological approach is applied, taking into account in a simple form two obvious phenomena: the contact of a microorganism with a substrate molecule and the inhibitory effect of the medium on it. The numerical values of empirical dependency coefficients for the studied processes are calculated. A differential equation is proposed at the macro level that describes the kinetics of biochemical destruction. The concept of a macrokinetic mathematical model of bioremediation is defined as a system of two functions that quantitatively reflect the dependence of the specific oxidation rate of pollution on its concentration and concentration on time, as well as satisfying the relationship between the relationships of the same parameters in differential form. The dependence of concentration on time is defined both in the form of a numerical integration algorithm and in the form of an approximate formula. The adequacy and universality of the proposed model for the studied processes is proved. The advantage of the proposed model of substrate inhibition kinetics is the simplicity of the structure of the basic formula and the ease of determining empirical coefficients based on this. In addition to numerical integration for determining the time of destruction, an approximate analytical solution is found, which can be adequately used in the concentration range of the experimental study. Further research is aimed at developing methods for calculating non-stationary processes in biochemical purification plants of certain specific types.

The finite amount of energy carriers affects all of us. It is important to utilize all available sources and to find new sources of energy. The confectionery industry generates both solid and liquid waste during the production of confectioneries, which could be utilized as a substrate for biogas production. However, problems might arise during the biogas process since these kinds of waste are very rich in carbohydrates. The initial breakdown of the substrate would probably cause an accumulation of fermentation products such as volatile fatty acids (VFA) and a low pH. A solution to this might be to use a two-stage process. The first stage would be a pre-fermentation that should be optimized for production of fermentation products such as ethanol and VFA. The aim of this master thesis is to evaluate the biogas potential of confectionery waste products. The confectionery waste products are evaluated using a continuous two-stage process, batch experiments and theoretical calculations of the methane potential. The potential from process wastewater was examined. Depending on COD reduction for a reactor and COD content of process wastewater, an annual amount of 75 000 m3 or of 857 000 m3 of process wastewater is necessary to produce enough biogas for a gas engine to continuously convert the biogas to electricity. A batch experiment evaluating the methane production potential of nine different confectionery waste products from a large confectionery industry gave a range of 430 - 690 NmL/g VS, which is relatively high. A continuous experiment in two lab-scale reactors with a HRT of two days worked satisfactory. The gas production was stable periodically with a carbon dioxide content above 60%. The pH was low (3.4 - 3.6) throughout the experiment for one of the reactors. However, addition of digester sludge from a methane-producing reactor towards the end of the experiment resulted in a higher pH and more VFA available for utilization in the second stage. The main fermentation products were: acetic acid, lactic acid, ethanol and carbon dioxide. A second batch experiment showed that the methane potential was not affected by pre-fermentation. A carbon balance calculation of the process indicates that 57% of the ingoing organic matter is fermented within only two days and ends up in the known fermentation products. The study shows that confectionery waste products are well suited for two-stage anaerobic digestion.<br>Den ändliga mängden av energibärare påverkar oss alla. Det är därför viktigt att utnyttja alla tillgängliga men även att finna nya energibärare. Konfektyrindustrin generar restprodukter (avfall) både i fast och flytande form, vilka båda kan utnyttjas för produktion av biogas. Det kan dock uppstå problem i biogasprocessen eftersom dessa innehåller en stor mängd kolhydrater. Den initiala nedbrytningen kan ge upphov till en ackumulering av VFA och ett lågt pH. En lösning på detta problem kan vara att använda en två-stegs process. Första steget är en för-fermentering, som ska optimeras för att producera fermentationsprodukter så som etanol och VFA. Syftet med detta arbete är att utvärdera biogaspotentialen från konfektyrrestprodukter genom att använda en kontinuerlig två-stegs process, batchförsök och teoretiska beräkningar av metanpotentialen. Potentialen undersöktes från processvatten. Beroende på COD reduktion i en reaktor och COD innehåll i processvatten, är en årlig mängd av 75 000 m3 eller av 857 000 m3 processvatten nödvändig för att producera en tillräcklig mängd biogas så att en gasmotor kontinuerligt kan omvandla biogasen till elektricitet. Ett batchförsök med nio olika restprodukter från en storskalig konfektyrproducent visade en relativt hög metanpotential (430 - 690 NmL/g VS). Ett kontinuerligt reaktorexperiment genomfördes i laboratorieskala med två reaktorer, där uppehållstiden var två dagar. De två för-fermenteringsreaktorerna presterade tillfredsställande. Gasproduktionen var periodvis stabil med en koldioxidhalt över 60%. pH var lågt (3,4 och 3,6) genom hela experimentet för en av reaktorerna. För den andra reaktorn gjordes tillsatser av reaktormaterial från en metanproducerande reaktor i slutet av experimentet. Dessa tillsatser ökade pH och totalmängden av VFA, som kan utnyttjas i det andra steget. Huvudfermentationsprodukterna är acetat, laktat, etanol och koldioxid. Ytterligare batchförsök visade att för-fermentation inte verkar påverka metanpotentialen för konfektyrrestprodukter. En kolbalans av processen indikerar att 57% av ingående kol återfinns i de kända fermentationsprodukterna inom två dagar. Studiens resultat visar att avfallsprodukter från konfektyrindustrin lämpar sig väl för två-stegs anaerob rötning.

Biofuels produced from lignocellulosic biomass via the fermentation platform are sustainable energy alternatives to fossil fuels. Process Systems Engineering (PSE) uses computer-based tools and methods to design, simulate and optimize processes. Application of PSE tools to the design of economic biorefinery processes requires the development of simulation approaches that can be integrated with existing, mature PSE tools used to optimize traditional refineries, such as Aspen Plus. Current unit operation models lack the ability to describe unsteady state fermentation processes, link unsteady state fermentation with in situ separations, and optimize these processes for competing factors (e.g., yield and productivity). This work applies a novel architecture of commercial PSE tools, Aspen Plus and MATLAB, to develop techniques to simulate time-dependent fermentation without and with in situ separations for process design, analyses and optimization of the operating conditions. Traditional batch fermentation simulations with in situ separations decouple these interdependent steps in a separate “steady state” reactor followed by an equilibrium separation of the final fermentation broth. A typical mechanistic system of ordinary differential equations (ODEs) describing a batch fermentation does not fit the standard built-in power law reaction kinetics model in Aspen Plus. To circumvent this challenge, a novel platform that links the batch reactor to a FORTRAN user kinetics subroutine (incorporates the ODEs) combined with component substitution (to simulate non-databank components) is utilized to simulate an unsteady state batch and in situ gas stripping process. The resulting model system predicts the product profile to be sensitive to the gas flow rate unlike previous “steady state” simulations. This demonstrates the importance of linking a time-dependent fermentation model to the fermentation environment for the design and analyses of fermentation processes. A novel platform linking the genetic algorithm multi-objective and single-objective optimizations in MATLAB to the unsteady state batch fermentation simulation in Aspen Plus through a component object module communication platform is utilized to optimize the operating conditions of a typical batch fermentation process. Two major contributions are: prior concentration of sugars from a typical lignocellulosic hydrolysate may be needed and with a higher initial sugar concentration, the fermentation process must be integrated with an in situ separation process to optimize the performance of fermentation processes. With this framework, fermentation experimentalists can use the full suite of PSE tools and methods to integrate biorefineries and refineries and as a decision-support tool to guide the design, analyses and optimization of fermentation-based biorefineries.

<p>The present Master thesis describes the development and optimization of a fed-batch process for production of recombinant proteins in Escherichia coli BL21(DE3) in a multi-fermentor system. The system consists of six 1-liter fermentors, capable of producing 500-1500 μg/mL with present protocol.</p><p>Response surface methodology (RSM) was used for multivariable optimization regarding cultivation time, pH, temperature and feed rate. Optimal protein expression conditions were found out to be 17.8 h cultivation time, 36.7 ºC, pH 6.8 and a feed rate corresponding to specific growth of 0.23 h-1, on glucose substrate. The aggregation of expressed proteins to inclusion bodies, could not be affected by the various growth conditions employed during cultivations.</p><p>A study was conducted regarding growth conditions effect on phosphogluconoylation of expressed proteins. In ten fed-batch cultivations on glucose, LC/MS analysis showed a gluconoylated fraction with additional 178 Da mass, but no correlation between growth conditions and gluconoylation could be found. In two fed-batch cultivations on glycerol-feed, a lower feed rate resulted in no gluconoylation, while a higher did. An explanation would be that the lower amount of available intra-cellular carbon limits formation of gluconoylation precursors.</p>

L’intérêt pour la paroi de la levure s’est accru récemment par l’explosion des activités de bioraffineries augmentant la production de biomasse, et par le besoin de valoriser cette biomasse dans d’autres débouchés comme en nutrition animale et en œnologie pour leurs propriétés probiotiques et de sorption. Le but de cette thèse était de combiner des approches biochimiques, biophysiques et les puces à ADN afin d'étudier les relations entre ces paramètres ainsi que de mettre en évidence l'impact des souches, des conditions de croissance et des procédés sur la composition et les propriétés biophysiques de la paroi cellulaire. Une méthode acido-enzymatique a été développée pour quantifier spécifiquement chacun des quatre composants de la paroi cellulaire de la levure, à savoir les mannanes, la chitine, les β-1,3-glucanes et les β-1,6-glucanes. Cette méthode a été validée sur des souches mutantes et a permis d’évaluer les effets de divers stress. Ultérieurement, l'utilisation de la microscopie à force atomique (AFM) a permis l'étude des mêmes souches et de quatre souches utilisées dans la fermentation industrielle. Ils ont démontré des propriétés nanomécaniques et adhésives distinctes, en raison de différences dans la composition et la structure de la paroi cellulaire. Dans la dernière partie, les effets du procédé d’autolyse et du séchage à lit fluidisé sont présentés. Ce procédé industriel ne modifie pas la composition de la paroi cellulaire, mais induit une modification de la topographie et des propriétés de surface de la cellule. En outre, en utilisant l'AFM nous avons imagés sur S. cerevisiae des patchs hautement adhésifs formant des nanodomaines à la surface de la cellule<br>Due to increasing yeast biomass production resulting from the expansion of the Biorefinery as an alternative to petrol-based energy, the yeast cell wall is receiving an increasing interest as an added value product targeting agro-nutrition markets, such as in animal nutrition and in wine for its probiotic and sorption properties. The purpose of this thesis was therefore to combine DNA microarrays, biochemical and biophysical approaches in order to investigate the relationships between these parameters as well as to highlight the impact of strains, growth conditions and processes on the cell wall composition and biophysical properties. To achieve this objective, an acido-enzymatic method was developed to specifically quantify each of the four components of the yeast cell wall, namely mannan, chitin, β-1,3-glucan and β-1,6-glucan. This method was validated on mutant strains and allowed highlighten various stresses effects. Then, the use of atomic force microscopy (AFM) has allowed investigating the same strains and four strains used in industrial fermentation. They demonstrated distinct nanomechanical and adhesive properties, due to differences in their cell wall structure and composition. In the last part, the effects of the autolysis and fluid-bed drying processes are presented. This industrial process does not change the composition of the cell wall but induces a modification in topography and surface properties of the cell. Moreover, using AFM we imaged on S. cerevisiae cell surface highly adhesive patches forming nanodomains

The overall objective of this work has been to adopt new developments and techniques in the area of measurement, modelling and control of fermentation processes. Flow cytometry and software sensors are techniques which were considered ready for application and the focus was set on developing tools for research aiming at understanding the relationship between measured variables and process quality parameters. In this study fed-batch cultivations have been performed with two different strains of Escherichia coli (E.coli) K12 W3110 with and without a gene for the recombinant protein promegapoietin. Inclusion body formation was followed during the process with flow cytometric detection by labelling the inclusion bodies with first an antibody against the protein promegapoietin and then a second fluorescent anti-antibody. The approach to label inclusion bodies directly in disintegrated and diluted cell slurry could be adopted as a method to follow protein production during the process, although the labelling procedure with incubation times and washings was somewhat time-consuming (1.5 h). The labelling of inclusion bodies inside the cells to follow protein production was feasible to perform, although an unexplained decrease in the relative fluorescence intensity occurred late in process. However, it is difficult to translate this qualitative measurement into a quantitative one, since a quantitative protein analysis should give data proportional to the volume, while the labelling of the spheric inclusion bodies gives a signal corresponding to the area of the body, and calibration is not possible. The methods were shown to be useful for monitoring inclusion body formation, but it seems difficult to get quantitative information from the analysis. Population heterogeneity analysis was performed, by using flow cytometry, on a cell population, which lost 80-90% viability according to viable count analysis. It was possible to show that the apparent cell death was due to cells incapable of dividing on agar plates after induction. These cells continued to produce the induced recombinant protein. It was shown that almost all cells in the population (≈97%) contained PMP, and furthermore total protein analysis of the medium indicated that only about 1% of the population had lysed. This confirms that the "non-viable" cells according to viable count by cfu analysis produced product. The software sensors XNH3 and µNH3, which utilises base titration data to estimate biomass and specific growth rate was shown to correlate well with the off-line analyses during cultivation of E. coli W3110 using minimal medium. In rich medium the µNH3 sensor was shown to give a signal that may be used as a fingerprint of the process, at least from the time of induction. The software sensor KLaC* was shown to respond to foaming in culture that probably was caused by increased air bubble dispersion. The RO/S coefficient, which describes the oxygen to substrate consumption, was shown to give a distinct response to stress caused by lowered pH and addition of the inducing agent IPTG. The software sensor for biomass was applied to a highly automated 6-unit multi-bioreactor system intended for fast process development. In this way also specific rates of substrate and oxygen consumption became available without manual sampling.<br>QC 20100819

Orientador: Edson Rosa Pimentel<br>Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia<br>Made available in DSpace on 2018-08-26T20:21:48Z (GMT). No. of bitstreams: 1 Almeida_MarcosdosSantosde_D.pdf: 2549790 bytes, checksum: f7c5cf41c1fe21e8441475d830717187 (MD5) Previous issue date: 2015<br>Resumo: Vários estudos indicam que a acupuntura (AC) tem efeito anti-inflamatório (AI) sistêmico e mecanotransdutor local. Estes efeitos podem potencialmente acelerar o processo de cicatrização. Portanto, o objetivo deste estudo foi analisar o efeito de diferentes protocolos de AC nas características bioquímicas, organizacionais e ultraestruturais de tendões de ratos durante o processo de cicatrização. Ratos Wistar machos com 60 dias de idade foram divididos nos seguintes grupos: grupo não tenotomizado ou normal (N), grupo tenotomizado (T), tenotomizado e submetido à AC no ponto E-36 (E36), tenotomizado e submetido à AC no B-57 (B57), tenotomizado e submetido à AC manual no ponto E-36 e B-57 (EB) e tenotomizado e submetido à eletroacupuntura (EA) nos dois pontos citados. Os animais foram submetidos à eutanásia nos dias 7, 14 e 21 após a lesão. Em seguida, os tendões calcâneos foram coletados, processados e analisados para quantificação de proteínas não colagênicas através do método de Bradford. Para a análise organizacional, foi utilizada a quantificação da birrefringência das fibras de colágeno através da microscopia de polarização. Na analise ultraestrutural foram mensurados os diâmetros das fibrilas de colágeno e em seguida foram construídos histogramas de distribuição destes diâmetros assim como foi calculado o diâmetro da massa-média - MAD - das fibrilas de colágeno. Os resultados mostraram que o uso associado dos pontos E-36 e B-57 aumenta tanto a birrefringência das fibras de colágeno nos dias 14 e 21 após a lesão quanto o MAD nos três períodos analisados. A reorganização das fibrilas de colágeno também aumentou no dia 21 após a lesão com a aplicação dos dois pontos citados. No entanto, a aplicação da EA não promoveu aumento da birrefringência ou da reorganização das fibrilas assim como diminuiu o MAD no dia 21 após a lesão. Desta forma, concluímos que o uso da AC manual nos pontos E-36 e B-57 tem efeito no restabelecimento das propriedades estruturais e ultraestruturais do colágeno de tendões durante o processo de cicatrização. Estes resultados sugerem o fortalecimento da estrutura tendínea com consequente aumento da resistência a re-ruptura e o potencial uso da AC em protocolos de reabilitação<br>Abstract: Several studies indicate that acupuncture (AC) has both local mechanotransductor and systemic anti-inflammatory (AI) effects. These effects could potentially accelerate the healing process. Therefore, the aim of this study was to analyze the effect of different protocols of AC on the biochemical, organizational and ultrastructural aspects of rat tendons during the healing process. Male Wistar rats at 60 days of age were divided into the following groups: not tenotomized or normal group (N), tenotomized group (T), tenotomized and submitted to AC at ST-36 point (ST36), tenotomized and submitted to AC at BL-57 point (BL57), tenotomized and submitted to manual AC at ST-36 and BL-57 points (SB) and tenotomized and submitted to electroacupuncture (EA) on the two points mentioned. The animals were euthanized on 7th, 14th and 21th days after injury. Thereafter, the Achilles tendons were collected, processed and analyzed to quantify non-collagenic proteins using the method of Bradford. For organizational analysis, it was quantified the collagen fibers birefringence by polarizing microscopy. In the ultrastructural analysis were measured the diameters of collagen fibrils and then histograms of distribution of these diameters were built as well as mass-average diameter - MAD - of collagen fibrils was calculated. The results showed that the associated use of ST-36 and BL-57 points increases both the birefringence of collagen fibers at 14th and 21th days and the MAD at the three periods analyzed. The reorganization of collagen fibrils also increased at 21th day with the application of the two points mentioned. However, the application of EA did not increase the birefringence or the reorganization of fibrils as well as decreased the MAD of fibrils at 21th after injury. Thus, we conclude that the use of manual AC at ST-36 and BL-57 points is effective in restoring the structural and ultrastructural properties of the collagen of tendons during the healing process. These results suggest strengthening the tendon structure with consequent increased resistance to re-rupture and the potential use of AC in rehabilitation protocols<br>Doutorado<br>Anatomia<br>Doutor em Biologia Celular e Estrutural

Nanofiltration is a proper way of preparation of drinking water. The process has a lot in common with reverse osmosis where a semipermeable membrane is used to separate a pure water flow from the remaining water. A pilot plant was used to ensure that nanofiltration is a suitable technology for a specific waterworks station in Glemmingebro, Ystad. The purpose was also to find out if a pH-adjustment is needed for feed water in a possible full scale plant.  In this case nanofiltration is used mainly to remove sulphate and calcium since the raw water contains concentrations of these substances which exceed the limits for potable water. It is important to receive a high recovery (RC), otherwise the deliviery of water won’t be high enough. On the other hand a high RC increases the risk of precipitates, which require more frequent cleaning. An optimal RC has already been calculated, the intention of this survey is to consider whether the theory is consistent with reality or not. Calculations of the highest acceptable pH-value were made in a computer program from Dow, ROSA. The result indicates that nanofiltration is suitable to use in this waterworks station and a pH-adjustment is needed when the pH level of raw water exceeds 7.7.<br>Tjänligt dricksvatten är en självklarhet för många. Processen som krävs för att producera vattnet är emellertid ganska komplicerad. Vattentäkterna har varierande förutsättningar och problematiken för varje enskilt vattenverk löses därför specifikt. En teknik som kan användas vid dricksvattenberedning är membranteknik i form av nanofiltrering. För att en membranprocess ska fungera krävs ett semipermeabelt membran vilket innebär att endast vissa molekyler tillåts att passera. Detta fenomen kallas omvänd osmos. Vattenmolekylerna diffunderar mot koncentrationsgradienten med hjälp av energi i form av ett högt tryck. På så vis återfås ett rent vatten från ett vatten innehållande en högre andel lösta ämnen. Ett av de centrala begreppen inom membranteknik är recovery (RC). RC (%) = permeatflöde/inflöde x 100 Permeat är den del av vattnet som filtrerats genom membranet, som senare ska bli dricksvatten. Enheten för recovery är %. Inflödet är den totala mängd vatten som matas in i anläggningen. Det vattnet som inte filtreras igenom membranet bildar tillsammans med de avskilda ämnena det flöde som kallas koncentrat eller avloppsflöde. En hög recovery innebär att av det vattnet som matas in till anläggningen går en större del ut som dricksvatten och en mindre del till avlopp. En låg recovery medför att det behövs en större mängd matarvatten för att erhålla samma mängd permeat. Med ökad recovery ökar koncentrationen av de avskilda ämnena i avloppsflödet. Detta sker eftersom samma mängd ämnen avskiljs, men volymen vatten till avlopp minskar. Löslighetsprodukten för svårlösliga föreningar riskerar då att överskridas, vilket resulterar i att de fälls ut och orsakar igensättning av membranet.  För att motverka igensättning kan en antiscalent användas. Optimal recovery är högsta möjliga, som inte orsakar igensättning av membranet. Den här studien är utförd på en specifik anläggning för Ystad kommun. Vattenverket är beläget i Glemmingebro. Invånarna i östra delen av kommunen får sitt vatten från kommunens största vattenverk, men det blandas upp med vatten från det lokala verket.  Sommartid råder det brist på vatten och planen är att hushållen i Glemmingebro i framtiden ska få större delen av sitt vatten från det lokala verket under den här perioden. Förutsättningarna för den här grundvattentäkten är att vattnet innehåller höga halter av både kalcium och sulfat. Det krävs en process där båda dessa ämnen reduceras för att uppnå tjänligt dricksvatten. Den höga halten av kalcium bidrar till att råvattnets hårdhetsgrad är 21dhᵒ. Sweco har gjort inledande undersökningar, där de kommit fram till att membranteknik troligtvis är lämpligt att använda vid beredning av dricksvatten i Glemmingebro. De har hyrt in två pilotanläggningar för vidare försök. Syftet med denna undersökning är att genom utredning av pilotförsök säkerställa att membranteknik är lämpligt att använda för dricksvattenberedning på Glemmingebro vattenverk, samt att utvärdera om det behövs en pH-sänkning för inkommande råvatten. Jag har också för avsikt att skapa en rapport för de som vill veta hur tekniken fungerar, alternativt är intresserade av dricksvattenberedning. Min förhoppning är att jag ska kunna belysa vad som är extra tänkvärt vid ett pilotförsök inom membranteknik, vilka problem som uppstår och hur dessa löses. Resultatet visar att membranteknik lämpar sig för beredning av dricksvatten på Glemmingebro vattenverk. Med en recovery på 75 % bedöms intervallen mellan tvätt av membranet överstiga fyra veckors normal drift vilket är önskvärt för anläggningen.  Avskiljningsgarden för oönskade ämnen har i pilotförsöken uppnått 95 % (kalcium), respektive 98 % (sulfat), vilket är förväntad nivå. Råvattnets pH varierar mellan 8,4 och 7,1. Analysdata visar att de högst uppmätta värdena främst förekommit sommartid. Under den här perioden antas råvattentemperaturen vara 15ᵒC. PH för inkommande vatten behöver vid denna temperatur regleras då pH överstiger 7,7. Detta för att undvika igensättning av membranet. Viktigt att notera är att en pilotanläggning inte visar hela verkligheten, utan endast ger en ungefärlig uppskattning om vad utfallet blir.<br>ombyggnad av Glemmingebro vattenverk

The American Cancer Society predicts that 1,665,540 people will be diagnosed with cancer, and 585,720 people will die from cancer in 2014. One of the most common types of cancer in the United States is skin cancer. Melanoma alone is predicted to account for 10,000 of the cancer related deaths in 2014. As a highly mobile and aggressive form of cancer, melanoma is difficult to fight once it has metastasized through the body. Early detection in such varieties of cancer is critical in improving survival rates in afflicted patients. Present methods of detection rely on visual examination of suspicious regions of tissue via various forms of biopsies. Accurate assessment of cancerous cells via this method are subjective, and often unreliable in the early stages of cancer formation when only few cancer cells are forming. With fewer cancer cells, it is less likely that a cancer cell will appear in a biopsied tissue. This leads to a lower detection rate, even when cancer is present. This lack of detection when cancer is in fact present is referred to as a false negative. False negatives can have a highly detrimental effect on treating the cancer as soon as possible. More accurate methods of detecting cancer in early stages, in a nonsubjective form would alleviate these problems. A proposed alternative to visual examination of biopsied legions is to utilize fluorescent nanocrystalline biomarker constructs to directly attach to the abnormal markers found on cancerous tissues. Quantum dots (QDs) are hydrophobic nanoscale crystals composed of semiconducting materials which fluoresce when exposed to specific wavelengths of radiation, most commonly in the form of an ultraviolet light source. The QD constructs generated were composed of cadmium-selenium (CdSe) cores encapsulated with zinc-sulfide (ZnS) shells. These QDs were then encapsulated with phospholipids in an effort to create a hydrophilic particle which could interact with polar fluids as found within the human body. The goal of this thesis is to develop a method for the solubilization, encapsulation, and initial functionalization of CdSe/ZnS QDs. The first stage of this thesis focused on the generation of CdSe/ZnS QDs and the fluorescence differences between unshelled and shelled QDs. The second stage focused on utilizing the shelled QDs to generate hydrophilic constructs by utilizing phospholipids to bind with the QDs. Analysis via spectroscopy was performed in an effort to characterize the difference in QDs both prior to and after the encapsulation process. The method generated provides insight on fluorescence trends and the encapsulation of QDs in polar substances. Future research focusing on the repeatability of the process, introducing the QD constructs to a biological material, and eventual interaction with cancer cells are the next steps in generating a new technique to target and reveal skin cancer cells in the earliest possible stages without using a biopsy.

Com o objetivo geral de elaborar e testar uma metodologia para comparar custos diretos unitários totais de análises bioquímicas quantitativas em processo manual e em processo automático no Laboratório de Análises Clínicas do Hospital Universitário da USP-LAC/HU e com os objetivos específicos de (a) aplicar a metodologia no LAC/HU, (b) avaliar se o auto-analisador era adequado à rotina, (c) qual o tempo teórico despendido por cada um dos processos - manual e automático - para realizar o total de análises requisitadas em 1989 e (d) proceder simulações do modelo de análises para situações hipotéticas a fim de verificar sua sensibilidade, foram estudados 7 tipos de análise - glicose, creatinina, uréia, sódio & potássio, ácido úrico, proteínas totais e cálcio. Adequou-se uma metodologia para avaliar o custo unitário direto total de cada um e conhecer qual processo apresentava os menores valores; os 7 tipos estudados foram submetidos a ambos no decorrer da pesquisa em 1989. Procurou-se, também, constatar se o equipamento utilizado no processo automático era adequado à rotina do HU e determinar o tempo teórico despendido por ambos os processos na execução de cada tipo de análise. O número estudado de análises foi determinado estatisticamente, o tempo de mão-de- obra gasto na execução das análises foi cronometrado, os dados referentes ao material de consumo, manutenção e depreciação foram obtidos nos respectivos processos de licitação e/ou aquisição. Para determinar os diversos custos unitários diretos totais foram selecionados os custos relevantes dos dois processos de execução e os resultados obtidos foram comparados. Verificou-se que os custos unitários diretos totais do processo manual foram menores que os do processo automático, exceto aqueles referentes à análise de sódio & potássio. Foi constatado, igualmente, pela comparação do tempo teórico despendido, se as análises dos 7 tipos fossem realizadas por ambos os processos, que o processo automático teria gasto um tempo menor que o processo manual na realização das duas etapas. Como os resultados encontrados divergiram dos esperados, pois a maioria dos custos unitários diretos totais do processo manual foi menor que os do processo automático, a pesquisa foi ampliada para serem acrescentadas 5 situações hipotéticas nas quais se admitiu, primeiramente, que dos 7 tipos de análises mencionados foram realizadas (a) 28.509 análises (número total das glicemias, o tipo mais requisitado) e (b) 47.894 análises (capacidade operacional do autoanalisador); a seguir admitiu-se que outros 4 tipos de análises-colesterol, bilirrubina, ferro e triglicérides, programados no auto-analisador e cujas análises não foram realizadas, o teriam sido nas seguintes quantidades: (a) o número real de análises executadas em 1989 das 11 análises e, também, os dois números das duas primeiras hipóteses, isto é, (b) 28.509 análises (número total das glicemias, o tipo mais requisitado) e (c) 47.894 análises (capacidade operacional do autoanalisador). Realizada a comparação do custo unitário direto total de cada um dos processos verificou-se que o processo automático, mesmo utilizando o auto-analisador em suas capacidades quantitativa e qualitativa totais, teria para a maioria das análises um custo unitário direto total maior que o do processo manual.<br>Seven types of biochemical analyses were studied (glucose, creatinine, urea, sodium & potassium, uric acid, total proteins and calcium), in orden to design and test a methodology aiming the comparison of total unitary direct cost of quantitative biochemical analyses between a manual process and an automatic one in the Clinical Analysis Laboratory (LAC) of Hospital Universitário from the University of São Paulo (HU). The specific objectives were a) to aply the methodology in the LAC, b) to determine wether the autoanalyzer was adequate for routine work, c) to measure the theorical time spent for each one of the processes - manual and automatic - to satisfy the demand of all the analyses accomplished during 1989, and d) to simulate a model of analyses for hypothetical situations in order to test its sensitivity. A specific methodoly was designed in order to evaluate the total unitary direct cost of each of the biochemical tests and to find out which of them had the lowest values. A side observation dealt with the adequacy to HU routine of the equipment used for the automatic process. All studies were done in 1989. The sample was determined with statistical tools, the time of manual labor was chronometered and the data related to supplies, maintenance and depreciation were gathered from their bidding and/or acquisition processes. To obtain the different total unitary direct costs, a comparison among relevant costs from the processes was established. The total unitary direct costs from the manual processes were lower than the automatic ones, except for sodium & potassium. If all 7 types were done by both processes, the automatic process would have taken less time than the manual one. These findings were different from what was expected, since costs for manual processes were lower than those for automatic ones. Therefore, the research was redesigned, to add 5 new hypothetic situations: a) the 7 types equaled 28509 analyses (total of glukemia tests, the one with the largest demand); b) the 5 types equaled 47894 analyses (operational capacity of the autoanalyzer) and c) under the false assumption that cholesterol, bilirubina, iron and triglycerides were done using autoanalyzer, three situations were simulated: (a) the actual number of analyses done in 1989; (b) 28509 analyses (see a above) and (c) 47894 (see b above). Comparing the total unitary direct cost of each of the processes, it was observed that for the automatic process, even using the autoanalyzer in full capacity (quantitative and qualitative), it would be greater than for the the manual process, for most cases.

Wastewater treatment facilities around the world use significant amount of energy which contributes to large quantities of greenhouse gas (GHG) emissions. According to the U.S.EPA, nearly 3% of the USA's energy is used to treat wastewater. This consumption is increasing at faster rates with increase in population and regulations. Wastewater facilities use large number of pumps in their transfer stations, treatment plants, and effluent pump stations. All these pumps consume considerable amounts of energy. This study presents a preliminary energy inspection of two facilities from Louisiana. This audit provides an inventory of the energy consumed for various activities like pumping, treatment, and discharge. This analysis helps the operators to identify the potential power consuming areas and optimize by adopting several energy conservation measures (ECMs). This study also involves the quantification of GHG emissions based on the energy consumption. The benefits of the study include minimizing energy and GHG emission.

Fossil based energy resources are dominating the world’s primary energy consumption for the last century. However, with decreasing crude oil reservoirs and the role they play in global warming by emitting greenhouse gases, the focus has been turned towards improved utilization of renewable resources and the need for new, sustainable fuels and chemicals is more urgent than ever. Biomass is a carbon neutral resource that can be used to produce biofuels and other useful chemicals. One such chemical is 1-butanol (or simply butanol), which has great potential as a gasoline substitute because of its favorable fuel properties. Butanol can be produced from acetone, butanol and ethanol (ABE) fermentation using e.g. Clostridium acetobutylicum. However, the concentration of butanol in fermentation in the resulting broth is limited to ca. 20 g/L due to its toxicity for microorganisms. Butyric acid is a precursor to butanol, which is produced prior to butanol in ABE fermentation. Butyric acid is an important industrial chemical, which can be further converted into a number of commercial compounds e.g. acetate butyrate, butyl acetate and butanol. Arginine is a semi-essential amino acid that has vast applications in the field of pharmaceutical and food industry. In addition, arginine can replace inorganic nitrogen as nitrogen source in fertilizers. It can be produced via fermentation of sugars using engineered microorganism like E. Coli, but like butanol its concentration is restricted to approximately 12 g/L. Due to low concentration of these useful chemicals in the resulting fermentation broths recovery of these chemicals remain challenging with today’s options and therefore  novel recovery process should be developed. In this study, zeolite adsorbents were used to recover butanol, butyric acid and arginine from model and real fermentation broths. Zeolite MFI adsorbent efficiently adsorbed butanol from model solutions with a saturation loading of 0.11 g/g- zeolite. On the other hand, adsorption of butyric acid was found to be strongly pH dependent, with high adsorption below and little adsorption above the pKa value of the acid. A structured adsorbent in the form of steel monolith coated with a silicalite-1 film was also used and performance was evaluated by performing breakthrough experiments at room temperature using model ABE fermentation broths and the results were compared with those obtained using traditional adsorbent sin the form of beads. Desorption studies showed that a high quality butanol product with purity up to 95.2% for butanol-water system and 88.5% for the ABE system can be recovered with the structured silicalite-1 adsorbent. Further, zeolite X adsorbents in the form of powder and extrudates was used to recover arginine from a real fermentation broth and also from aqueous model solutions. To the best of our knowledge, this is the first time recovery of arginine from real fermentation broths using any type of adsorbent is reported. Arginine loading of 0.15 g/g was observed at pH 11 using zeolite X powder. The selectivity for arginine over ammonia and alanine from the fermentation broth at pH 11 was 1.9 and 8.3, respectively, for powder and 1.0 and 4.1, respectively, for extrudates. Synthesis gas (CO + H2) can be produced e.g.by gasification of lignocellulose biomass. This synthesis gas can be used to produce methanol, which subsequently may be converted into gasoline using zeolite ZSM-5 catalyst. However, during Methanol to Gasoline (MTG) process, undesirable carbon residue (coke) is formed that gradually reduces the activity of catalyst. It was hypothesized that intracrystalline defects in the zeolite formed during conventional synthesis may accelerate the deactivation rate by coke formation. In this work, a novel ZSM-5 zeolite catalyst essentially free of intracrystalline defects was synthesized and evaluated in the  MTG reaction,. The novel catalyst showed significantly higher resistance towards deactivation by coke formation as compared to a reference catalyst containing defects.

No presente trabalho buscaram-se formas adequadas de produzir micélio de Ceriporiopsis subvermispora em culturas submersas, com o objetivo de preparar inóculo destinado ao processo de biopolpação. A obtenção de inóculo pode ser considerada uma etapa chave na biopolpação, visto que atualmente não existem processos industriais estabelecidos para a produção de micélio de basidiomicetos em quantidades compatíveis com a demanda prevista no processo. Mesmo uma planta industrial pequena de biopolpação (200 ton de polpa/dia) pode demandar cerca de 1 kg de micélio seco/dia, considerando-se as cargas de inoculação de 5 g de micélio seco/ton de madeira tratada. Desta forma, o sucesso desse processo pode depender de um baixo custo da produção dos inóculos fúngicos em larga escala. Os experimentos realizados consistiram inicialmente no crescimento do fungo em Erlenmeyers de 250 mL contendo 20 mL de meio líquido. Numa segunda etapa, os experimentos foram conduzidos em biorreatores de 1,5 L (Bioflo-NewBrunswick) e por último em um biorreator de 14 L. No último caso, o biorreator em questão foi especialmente desenhado e construído com o intuito de minimizar efeitos de cisalhamento da hifa durante o cultivo, mantendo ainda níveis adequados de aeração. Em Erlenmeyer foram feitos cultivos utilizando meio composto por 2,4% de extrato de batata e 0,7% de extrato de levedura (DB-EL), além de um meio potencialmente de menor custo que era composto por 2,0% de sacarose e 3,2% de milhocina (SM). A biomassa total obtida nos cultivos com meio SM foi compatível com aquela obtida no meio DB-EL. Os cultivos subsequentes realizados em biorreator utilizaram então o meio SM. O maior nível de biomassa fúngica obtida nos cultivos com biorreator agitado por pás (1,5 L) foi de 7,0 g/L, após 12 dias de crescimento. As maiores produtividades foram observadas a partir do 7o dia de cultivo, atingindo 0,93 g/L.dia quando a biomassa total era de 6,5 g/L. No biorreator de 14 L foram realizados 3 cultivos em condições diferenciadas (denominados de A, B e C). No cultivo A não houve intervenção em relação à correção de pH ou nível de nutrientes disponíveis ao longo de 14 dias. No cultivo B foi realizado o controle do pH a fim de mantê-lo entre 4,0 e 5,0. No cultivo C foi realizada a adição de sacarose correspondente a uma reposição de 5 g/L após 5 dias de crescimento. Os cultivos foram monitorados quanto à biomassa produzida, o pH, o teor de açúcar residual e o nível de O2 dissolvido. De forma geral, o acúmulo de biomassa sempre foi seguido pelo consumo de açúcares, pela diminuição nos níveis de O2 dissolvido e pela tendência de elevação do pH. A maior quantidade de biomassa foi obtida no cultivo C (14,1 g/L), correspondendo a uma produtividade de 1,72 g/L.dia. O surgimento de clamidósporos foi verificado em todos os tipos de cultivo, sendo que no cultivo C (em biorreator de 14 L) foi observada a menor quantidade de clamidósporos. Isso sugeriu que nessa situação houve melhores condições de crescimento para o fungo e menor estresse nutricional ou induzido por cisalhamento. A viabilidade dos inóculos preparados para a efetiva colonização da madeira foi testada e indicou que o inóculo produzido no biorreator de 14 L, tanto micélio como clamidósporos, foi efetivo para colonizar Eucalyptus grandis.<br>The present work evaluated suitable systems for producing Ceriporiopsis subvermispora mycelium in submerged cultures. Mycellium produced was used in subsequent inoculation of wood chips in biopulping processes. The development of appropriated technologies for producing large amounts of inoculum may be a key step in biopulping, since currently there are no established processes designed to produce basidiomycetes mycelium to supply biopulping demands. Even a small biopulping plant (200 ton pulp / day) may require about 1 kg of dry mycelium/day, taking into account inoculation of 5 g of dry mycelium/ton of wood to be biotreated. Thus, the success of this process may depend on the low cost of large scale fungal inoculum production. The experiments were conducted initially in 250 mL Erlenmeyer flasks containing 20 mL of liquid medium. In a second step, the experiments were conducted in bioreactors of 1.5 L (Bioflo-NewBrunswick) and finally in a bioreactor of 14 L. In the latter case, the bioreactor has been designed and constructed in order to minimize the effects of shear stress during cultivation and supply suitable aeration levels. Culture broths were composed of 2.4% potato extract and 0.7% yeast extract (BD-EL) or 2.0% sucrose and 3.2% of corn steep liquor (SM) that represent a potentially lower cost culture broth. The total biomass obtained in flask-cultures with SM broth was similar to that obtained with DB-EL. Cultures performed in the subsequent bioreactor used then the SM broth. The higher level of fungal biomass obtained in the cultures performed in the 1.5 L-stirred bioreactor was 7.0 g/L after 12 days. The highest mycelium yield was observed from the 7th day of cultivation, reaching 0.93 g/L.day and 6.5 g/L total biomass. In the 14 L-bioreactor, 3 experiments were performed in different conditions (called A, B and C). In cultivation A, no interventions were performed to correct the culture pH or the level of available nutrients over 14 days of culturing. In culture B pH was controlled to keep it between 4.0 and 5.0. Cultivation C was carried out with addition of sucrose (final concentration of 5 g/L) after 5 days of growth. All cultures were monitored with basis on the biomass produced, culture pH, residual sugar content and dissolved O2 levels. In general, biomass accumulation was always followed by sugar consumption, decrease in dissolved O2 levels and a rising tendency for the pH values. The highest biomass amount was obtained in culture C (14.1 g / L), corresponding to a yield of 1.72 g/L.day. The presence of clamydospores was observed in all cultures, whereas in culture C (in 14 L bioreactor) it appeared at the lowest amounts. This fact suggested that culture C presented the lowest stress level for the fungus, including low hypha shear stress and low nutritional depletion. The viability of inoculum prepared, both mycelium and clamydospores, was checked by culturing them on Eucalyptus grandis wood chips. Both inoculum types (prepared from the 14 L-bioreactor) were efficient on colonizing wood chips and to produce the manganese peroxidase enzyme.

Utilizou-se um meio de cultivo a base de farinha de mandioca, suplementado com nutrientes, em fermentador agitado (700 rpm) e aerado (10 litros de ar/min), com volume de reação de 10 litros praticamente constante, fração de inóculo de 10% em volume, ph 4,0 e temperatura de 35ºC. Foram realizados ensaios com concentração total de açúcares de 20 g/l e 40 g/l, tanto descontínuos como descontínuos alimentados. Nestes variou-se a vazão mássica de alimentação (fs), o instante de início de alimentação e a condição do xarope de farinha (previamente hidrolisado ou não). Repetições dos ensaios descontínuos indicaram variabilidade de resultados elevada. Não se observou expressivas mudanças no crescimento microbiano, a não ser pelo aumento na velocidade específica nos ensaios descontínuos alimentados a 20 g/l. A síntese de glicoamilase foi sensivelmente aumentada nos ensaios descontínuos alimentados a 20 g/l (produtividade dobrada). A 40 g/l, obteve-se produtividade 26% superior. Os melhores resultados foram obtidos com fs=17,1 gart/h a so=20 g/l e fs=32,2 gart/h a 40 g/l, e obteve-se o pior no ensaio em que se alimentou desde o início de cultivo. A so=20 g/l a repressão se apresenta como principal mecanismo de controle de síntese de glicoamilase, não ocorrendo a mesma a 40 g/l, ensaios nos quais a indução tornou-se muito relevante.<br>In order to study different processes and the influence of control mechanism on glucoamylase synthesis, several batch and fed-batch runs were made with Aspergillus awamori NRRL 3112. A medium containing cassava flour and nutrients were used in a 10 liters stirred and aerated tank, at pH 4,0 and temperature 35 °C. The batch and fed-batch runs used 20 and 40 g of total reducing sugars (TRS) per liter. In the fed-batch runs, the carbon source feed rate (fs), the feeding start time, and whether the syrup were pre-hydrolyzed or not were varied. Repeated batch runs showed significant variability. Notable changes in cell growth were not observed, unless by the increase of the specific growth rate in the 20 gTRS/l fed-batch runs. The enzyme productivity doubled in the lower sugar concentration fed-batch runs, but increased just 26% in the runs with 40g/l of TRS. The best results were achieved at 20g/l with carbon source feed rate=17,1 gTRS/h and fs=32,2 gTRS/h at 40g/l. The worst noted when the feeding started at the beginning of the run. At 20 gTRS/l, repression showed as the main mechanism control in order to synthesize glucoamylase. On the other hand induction became the relevant factor when 40gTRS/l were offered to microorganism.

Orientador: Marcus César Avezum Alves de Castro<br>Resumo: No Brasil são geradas quantidades significativas de resíduos orgânicos provenientes das perdas no segmento de produção e processamento da cadeia de fornecimento de alimentos, ou desperdiçadas no varejo e no consumo final. Ainda, no país há carência de aplicações do processo de digestão anaeróbia no tratamento dos resíduos orgânicos alimentares, tornando pertinente seu estudo. Este trabalho analisou a influência de diferentes proporções de resíduos alimentares no processo de biometanização. Os substratos consistiram de resíduos orgânicos coletados em restaurante universitário, sendo divididos em dois grupos: Resíduos de Pré-Preparo dos alimentos (RPP) e Resíduos de Sobras das refeições (RS). Testes em batelada foram realizados para avaliar o Potencial Bioquímico de Metano (BMP) das diferentes proporções de resíduos, sob condições mesofílica e termofílica, tendo como fontes de inóculos dois lodos anaeróbios granulares provenientes de reatores UASB (Upflow Anaerobic Sludge Blanket). Para verificar a atividade dos lodos foram realizados testes de Atividade Metanogênica Específica (AME), tendo o acetato como fonte de carbono, com aplicação de diferentes relações Substrato/Inóculo (So/Xo) (0,5, 1,0 e 1,5). Os resultados dos testes de AME sob condições mesofílica e termofílica demonstraram bom desempenho dos inóculos com índices 0,1965 e 0,1559 g DQOCH4.g STV-1.d-1, respectivamente, sendo que a relação So/Xo=1,0 foi a mais indicada para aplicação na biodigestão anaeróbia dos resíduo... (Resumo completo, clicar acesso eletrônico abaixo)<br>Abstract: In Brazil, significant amounts of organic waste are generated from losses in the production and processing segment of the food supply chain, or wasted on retail and final consumption. Still, in the country there is a lack of applications of the anaerobic digestion process in the treatment of organic food residues, making its study pertinent. This work analyzed the influence of different proportions of food residues on the biomethanization process. The substrates consisted of organic residues collected in a university restaurant, being divided into two groups: Food Pre-Preparation Residues (RPP) and Residues of Leftovers from meals (RS). Batch tests were carried out to evaluate the Biochemical Potential of Methane (BMP) of the different proportions of residues, under mesophilic and thermophilic conditions, having as inoculum sources two granular anaerobic sludge from UASB (Upflow Anaerobic Sludge Blanket) reactors. In order to verify the activity of the sludges, tests of Specific Methanogenic Activity (AME) were carried out, with acetate as carbon source, applying different Substrate/Inoculum (So/Xo) ratios (0.5, 1.0 and 1.5). The results of the AME tests under mesophilic and thermophilic temperatures showed good performance of the inoculum with indexes 0.1965 and 0.1559 g DQOCH4.g STV-1.d-1, respectively, with the ratio So/Xo=1.0 was the most indicated for application in anaerobic biodigestion of food waste. BMP tests under different proportions of food residues demonstrated ... (Complete abstract click electronic access below)<br>Mestre

A large part of energy carriers and chemicals of our society come from fossil fuels; due to their critical exploitation and environmental concerns, alternative solutions should be promoted. The goal of this work is to understand the environmental impacts of different supply chains of levulinic acid production. To enable this comparison, a cradle-to-gate LCA was conducted on different kinds of biomasses, under an innovative thermochemical process called Biofine. The study concerns biomass not only of agri-food origin (corn stover, barley straw, wheat straw), but also deriving from the waste collection supply chain (organic fraction of municipal solid waste). Results show that biochemicals production from waste-derived biomass represent the lowest environmental impact solution, compared to the supply chain with biomass of agri-food origin. For agri-food biomasses, the impacts deriving from the agricultural phase are orders of magnitude more significant than those of the industrial phase, while the transport phase is the least impacting in absolute terms. For waste-derived biomasses, transport is the most impacting phase of the entire supply chain.

The subject of this thesis is the generation of spatial temporal structures in living cells. Specifically, we studied the Min-system in the bacterium Escherichia coli. It consists of the MinC, the MinD, and the MinE proteins, which play an important role in the correct selection of the cell division site. The Min-proteins oscillate between the two cell poles and thereby prevent division at these locations. In this way, E. coli divides at the center, producing two daughter cells of equal size, providing them with the complete genetic patrimony. Our goal is to perform a quantitative study, both theoretical and experimental, in order to reveal the mechanism underlying the Min-oscillations. Experimentally, we characterize theMin-system, measuring the temporal period of the oscillations as a function of the cell length, the time-averaged protein distributions, and the in vivo Min-protein mobility by means of different fluorescence microscopy techniques. Theoretically, we discuss a deterministic description based on the exchange of Minproteins between the cytoplasm and the cytoplasmic membrane and on the aggregation current induced by the interaction between membrane-bound proteins. Oscillatory solutions appear via a dynamic instability of the homogenous protein distributions. Moreover, we perform stochastic simulations based on a microscopic description, whereby the probability for each event is calculated according to the corresponding probability in the master equation. Starting from this microscopic description, we derive Langevin equations for the fluctuating protein densities which correspond to the deterministic equations in the limit of vanishing noise. Stochastic simulations justify this deterministic model, showing that oscillations are resistant to the perturbations induced by the stochastic reactions and diffusion. Predictions and assumptions of our theoretical model are compatible with our experimental findings. Altogether, these results enable us to propose further experiments in order to quantitatively compare the different models proposed so far and to test our model with even higher precision. They also point to the necessity of performing such an analysis through single cell measurements.

The subject of this thesis is the generation of spatial temporal structures in living cells. Specifically, we studied the Min-system in the bacterium Escherichia coli. It consists of the MinC, the MinD, and the MinE proteins, which play an important role in the correct selection of the cell division site. The Min-proteins oscillate between the two cell poles and thereby prevent division at these locations. In this way, E. coli divides at the center, producing two daughter cells of equal size, providing them with the complete genetic patrimony. Our goal is to perform a quantitative study, both theoretical and experimental, in order to reveal the mechanism underlying the Min-oscillations. Experimentally, we characterize theMin-system, measuring the temporal period of the oscillations as a function of the cell length, the time-averaged protein distributions, and the in vivo Min-protein mobility by means of different fluorescence microscopy techniques. Theoretically, we discuss a deterministic description based on the exchange of Minproteins between the cytoplasm and the cytoplasmic membrane and on the aggregation current induced by the interaction between membrane-bound proteins. Oscillatory solutions appear via a dynamic instability of the homogenous protein distributions. Moreover, we perform stochastic simulations based on a microscopic description, whereby the probability for each event is calculated according to the corresponding probability in the master equation. Starting from this microscopic description, we derive Langevin equations for the fluctuating protein densities which correspond to the deterministic equations in the limit of vanishing noise. Stochastic simulations justify this deterministic model, showing that oscillations are resistant to the perturbations induced by the stochastic reactions and diffusion. Predictions and assumptions of our theoretical model are compatible with our experimental findings. Altogether, these results enable us to propose further experiments in order to quantitatively compare the different models proposed so far and to test our model with even higher precision. They also point to the necessity of performing such an analysis through single cell measurements.

Owing to depletion of fossil fuels, increase in crude prices, and large scale environmental pollutions forced to shift current research focus on renewable resources like biomass for production of fuels and chemicals. The bio-butanol is one such alternative fuel for application in existing internal combustion engines that can be produced by fermentation of biomass. The bio-butanol provides an alternative to butanol produced from petrochemical pathways. Thus there is a need to develop the process for large scale production of bio-butanol from biomass in cost effective manner. The objective of the present study is to design processes to produce bio-butanol from various feedstock including sugarcane, corn, and lignocellulosic biomass using aspen plus. The economic estimation of fixed capital investment and production costs has been carried out for a plant capacity of 10,000 tonne per year butanol. The yield of 0.39 g ABE/g glucose with ABE solvents in the ratio 3:6:1 has been considered in entire analysis. It has been found that the fixed capital investment for corn as feedstock was much higher compared to sugarcane and lignocellulosic biomass. This is because of the additional pretreatment required to extract starch from corn and medium preparation. Byproduct credits for gases and chemicals are taken into consideration to calculate the production cost of butanol. For a yield of 0.39 g ABE/g glucose, the bio-butanol production cost was estimated as $1.04, $1.89, $1.42 for sugarcane, corn, and lignocellulosic biomass respectively. These costs are sensitive to changes in feedstock cost which can change the butanol price significantly.

碩士<br>國立臺灣大學<br>環境工程學研究所<br>86<br>Abstract The main purpose of this study was to investigate the nutrient removal efficiency of Modified A/O System, and to analyze the mechanisms of biological nitrogen and phosphorus removal in this system by utilizing phosphorus fractionation method(the STS Method)、PHB and glycogen analysis . Experimental results indicated that, under different influent substrates and ORP control conditions, the TCOD removal efficiency of Modified A/O system achieved 90% and the TKN removal efficiency also achieved 95%. However, the TN removal efficiency could only maintain 50% and the TP removal efficiency only remained 20~30% .The results verified that NOx- N Loading in the anoxic tank influence the releasing of phosphorus magnificently and it *s difficult to treat nitrogen and phosphorus simultaneously Using the STS Method, we found that the amount of nucleic acid-P in anoxic tank rouse highly and this is due to the production of ATP by denitrification. The magnificent ATP would inhibit the polyphosphates breakdown reaction and thus inhibited the phosphorus releasing, this diminished the phosphorus treating ability of Modified A/O system. In addition, the analysis of PHB in anoxic sludge show that the fermentation of influent sugar-rich substrates poorly appeared, this may be the results of denitrification and cause the polyphosphates accumulating bacteria hardly to survive. By analyzing the relationship of the difference of ORP of two tanks and the phosphorus fraction absorption amount, we found that the major species supporting biological phosphorus removal were high molecular weight polyphosphates and protein- P. The nucleic acid-P had negative effect on biological phosphorus removal and was competitive to polyphosphates. Comparing this system with other systems revealed by literature, we found that the main factors causing poorly nutrient removal efficiency were the shortness of HRT and the failure of separation of phosphorus releasing and denitrification. The further study of elongation of HRT and the change of the microbe carrier materials in anoxic tank were suggested.

Some of the results presented in this document were submitted to the international journal of Phycological Research for publication and are currently under revision (Annex I).<br>The effect of drying on two brown seaweed (Cystoseira abies-marina, Cystoseira humilis) and two red seaweed species (Asparagopsis armata, Asparagopsis taxiformis), collected from the Azores Archipelago, was evaluated through the study of their proximate and mineral composition, relevant biological activities, such as antioxidant and anti-inflammatory properties, and significant bioactives, namely polyphenols and beta-glucans. Ash and protein content ranged from 25-56 g/100g dw and 3.5-13.1 g/100g dw, respectively. Sun-dried C. humilis had the highest moisture decrease. Concerning insoluble dietary fibre content, the genus Cystoseira presented superior concentrations (43.7-53.6 g/100g dw). Contrarily, the soluble dietary fibre content is superior in the Asparagopsis genus (8.0-13.2 g/100g dw). For the proximate composition, no significant differences were detected concerning the drying procedure. Seaweeds from the Cystoseira genus showed high polyphenol levels (176-678mg GAE/100 g dw), exceeding those determined in the Asparagopsis genus, regardless of drying process. This was partially reflected in the antioxidant activity, which showed that extracts from the Cystoseira species were often more antioxidant than those from Asparagopsis species. The influence of the drying technique upon the antioxidant activity was limited, since in many instances there was no effect. Concerning anti-inflammatory activity, in the case of shade-dried samples, C. humilis had a higher activity (>30% COX-2 inhibition) but was not rendered bioaccessible. Indeed, only A. taxiformis displayed anti-inflammatory activity in the bioaccessible fraction, leading to bioaccessibility factors in the 90-100% range. Therefore, though bioactivities were higher in the Cystoseira species, Asparagopsis species also had a positive bioactive potential. Sun-drying produced more negative effects than shade-drying, despite not being very extensive. Regarding elemental composition, iodine was present in a considerable amount in the Asparagopsis genus. Iron had high concentrations in the four species. Regarding contaminants, Cystoseira abies-marina showed high arsenic concentrations. Iodine, bromine, magnesium, and cadmium showed the highest bioaccessibility percentages.

Dissertation submitted in compliance with the requirements for the Master's Degree in Technology: Biotechnology, Technikon Natal, 2001.<br>Up until now extensive work has been done to develop kinetic models and related software that can be used successfully to simulate and design nitrification denitrification (ND) and nitrification denitrification biological excess phosphorus removal (NDBEPR) systems for efficient nitrogen removal. The denitrification kinetics of these systems have primarily been determined and attributed to the ordinary heterotrophic bacteria, now also known as the OHO fraction, otherwise not involved in biological excess phosphorus removal. However, denitrification kinetics determined for ND systems have been found to vary considerably at times when applied to NDBEPR systems because of varying OHO active fraction estimates and the unexplained occurrence of anoxic phosphorus removal and anysuccess achieved to date has been some what fortuitous. Ultimately variations in process performance and kinetics are attributable to inadequate control and lack of understanding of the ecological, physiological and biochemical activities of constituent microorganisms. There is growing concern and movement towards a better understanding of the microbial community within activated sludge in order to gain optimal control of the process.<br>M

Dissertação de Mestrado Integrado em Engenharia Química apresentada à Faculdade de Ciências e Tecnologia<br>Among all environmental problems faced nowadays, climate change is undoubtedly the most imminent. Biofuels such as biogas may have a major role in the replacement of fossil fuels. Biogas is the main product of anaerobic digestion (AD) and can be used for producing energy in an efficient and eco-friendly way. The main objective of this work is the study of the AD process to assess the potential valorization of agro-industrial wastes through experimental and theoretical biochemical methane potential (BMP) prediction. In a first phase, 40 scientific papers were analyzed and was possible to obtain the BMP for 149 substrates. Then, an exhaustive analysis of these data was carried out through simple linear and multivariate polynomial regressions. Moreover, experimental tests of BMP and AD batch tests were used to evaluate the methane production of three substrates, namely winery wastewater (WW), tomato waste (TW) and banana peel waste (BW). Substrate-inoculum (S/I) ratios were optimized for WW and then applied to other substrates. Through the explorative analysis of the literature data, it was possible to verify that the BMP is a very complex parameter, making its prediction very hard. In fact, the methods for predicting this parameter referred in several papers are unsatisfactory since large differences are found between the predicted and the experimental BMP values. With the multivariate polynomial regressions performed in this study, it was possible to develop two models that present great potential for BMP prediction. From experimental tests, it was possible to verify that TW is the substrate with the highest potential for AD, followed by BW and lastly, with a lower potential, WW. The batch AD of WW revealed that the optimal S/I ratio is 0.5, which is in accordance with the literature. By using this S/I ratio it was possible to obtain 358.6 NmL gVS-1 of biogas for WW, 453.4 NmL gVS-1 for TW and 574.1 NmL gVS-1 for BW.This study demonstrated that BMP plays a fundamental role in the evaluation of a substrate potential for AD. It was possible to conclude that this parameter may be assessed in a simpler and faster way by theoretical models when compared to the experimental laborious methodologies. It was also possible to show that the substrates explored in this study can be valorized through an AD process.<br>De entre todos os problemas ambientais enfrentados hoje em dia, as alterações climáticas são, sem dúvida, as mais preocupantes. Os biocombustíveis, como o biogás, começam a ter um papel importante na substituição dos combustíveis fósseis. Este gás é o principal produto da digestão anaeróbia (AD) e pode ser usado para produzir energia de forma eficiente e ecológica. Existem diversos tipos de substratos orgânicos biodegradáveis que podem ser utilizados como substrato para na AD. Portanto, é de grande importância avaliar a biodegradabilidade e o potencial bioquímico de metano (BMP) destes.Este trabalho tem como principal objetivo o estudo do processo de AD, a fim de avaliar a possibilidade de valorização de resíduos agroindustriais. Através da análise de 40 artigos científicos, foi possível obter o BMP para 149 substratos diferentes. Após esta coleta foi realizada uma análise exaustiva desses dados através de regressões lineares simples e regressões polinomiais multivariadas. Foram realizados testes experimentais de BMP e de AD para avaliar o potencial de produção de metano de três substratos, nomeadamente de um efluente vinícola (WW), resíduo de tomate (TW) e resíduo de cascas de banana (BW). A razão substrato-inoculo (S/I) foi otimizada através da digestão do substrato WW, sendo depois aplicada aos restantes.Através da análise exploratória dos dados da literatura, foi possível verificar se o BMP é um parâmetro bastante complexo, fazendo com que a sua previsão seja difícil. Daí, os métodos para prever esse parâmetro referidos na literatura serem insatisfatórios. Através das regressões polinomiais multivariadas realizadas neste estudo foi possível desenvolver dois modelos que apresentam grande potencial no que toca à previsão do BMP. A partir dos testes experimentais BMP, foi possível verificar que o TW é o substrato com o maior potencial de AD, seguido do BW e, por último, com menor potencial, o WW. A AD do WW revelou que a razão S/I ideal, dentro dos valores testados, é de 0.5. Através desta razão S/I, foi possível obter 358.6 NmL gVS-1 de biogás para WW, 453.4 NmL gVS-1 para TW e 574.1 NmL gVS-1 para o BW.Este estudo demonstrou que o BMP desempenha um papel fundamental na avaliação do potencial de um substrato para AD e que é possível prever este parâmetro de forma mais simples e rápida em comparação com o procedimento experimental. Foi possível também mostrar que os substratos explorados neste estudo podem ser valorizados através de um processo AD.