Bioprocess engineering of induced pluripotent stem cells for application in cell therapy and pre-clinical research

Dissertation to obtain Master Degree in Biotechnolog

Bioprocess Monitoring and Control

Process monitoring and control are fundamental to all processes; this holds especially for bioprocesses, due to their complex nature. Usually, bioprocesses deal with living cells, which have their own regulatory systems. It helps to adjust the cell to its environmental condition. This must not be the optimal condition that the cell needs to produce whatever is desired. Therefore, a close monitoring of the cell and its environment is essential to provide optimal conditions for production. Without measurement, no information of the current process state is obtained. In this book, methods and techniques are provided for the monitoring and control of bioprocesses. From new developments for sensors, the application of spectroscopy and modelling approaches, the estimation and observer implementation for ethanol production and the development and scale-up of various bioprocesses and their closed loop control information are presented. The processes discussed here are very diverse. The major applications are cultivation processes, where microorganisms were grown, but also an incubation process of bird’s eggs, as well as an indoor climate control for humans, will be discussed. Altogether, in 12 chapters, nine original research papers and three reviews are presented

Environmental biotechnologies for phosphogypsum recycling: protection of the atmosphere, hydrosphere and lithosphere

This monograph substantiates biotechnological solutions for recycling phosphogypsum together with other types of waste with the production of useful bio-based products in the framework of environmental protection technologies. The concept of the methodological approach to phosphogypsum utilisation in bioprocesses includes its integrated use in technologies for the protection of atmospheric air, hydrosphere, and lithosphere. The book is addressed to scientists, university professors, graduate, and postgraduate students, as well as to all those who are interested in the problem of waste recycling, in particular phosphogypsum, and the development of technological solutions for the protection of environmental components based on synergetic approach

Advances in Applied Biotechnology

Biotechnology is the scientific field of studying and applying the most efficient methods and techniques to get useful end-products for the human society by using viable micro-organisms, cells, and tissues of plants or animals, or even certain functional components of their organisms, that are grown in fully controlled conditions to maximize their specific metabolism inside fully automatic bioreactors. It is very important to make the specific difference between biotechnology as a distinct science of getting valuable products from molecules, cells or tissues of viable organisms, and any other applications of bioprocesses that are based on using the whole living plants or animals in different fields of human activities such as bioremediation, environmental protection, organic agriculture, or industrial exploitation of natural resources. The volume Advances in Applied Biotechnology is a scientific book containing recent advances of selected research works that are ongoing in certain biotechnological applications. Fourteen chapters divided in four sections related to the newest biotechnological achievements in environmental protection, medicine and health care, biopharmaceutical producing, molecular genetics, and tissue engineering are presented

Book of abstracts of the 10th International Chemical and Biological Engineering Conference: CHEMPOR 2008

This book contains the extended abstracts presented at the 10th International Chemical and Biological Engineering Conference - CHEMPOR 2008, held in Braga, Portugal, over 3 days, from the 4th to the 6th of September, 2008. Previous editions took place in Lisboa (1975, 1889, 1998), Braga (1978), Póvoa de Varzim (1981), Coimbra (1985, 2005), Porto (1993), and Aveiro (2001). The conference was jointly organized by the University of Minho, “Ordem dos Engenheiros”, and the IBB - Institute for Biotechnology and Bioengineering with the usual support of the “Sociedade Portuguesa de Química” and, by the first time, of the “Sociedade Portuguesa de Biotecnologia”. Thirty years elapsed since CHEMPOR was held at the University of Minho, organized by T.R. Bott, D. Allen, A. Bridgwater, J.J.B. Romero, L.J.S. Soares and J.D.R.S. Pinheiro. We are fortunate to have Profs. Bott, Soares and Pinheiro in the Honor Committee of this 10th edition, under the high Patronage of his Excellency the President of the Portuguese Republic, Prof. Aníbal Cavaco Silva. The opening ceremony will confer Prof. Bott with a “Long Term Achievement” award acknowledging the important contribution Prof. Bott brought along more than 30 years to the development of the Chemical Engineering science, to the launch of CHEMPOR series and specially to the University of Minho. Prof. Bott’s inaugural lecture will address the importance of effective energy management in processing operations, particularly in the effectiveness of heat recovery and the associated reduction in greenhouse gas emission from combustion processes. The CHEMPOR series traditionally brings together both young and established researchers and end users to discuss recent developments in different areas of Chemical Engineering. The scope of this edition is broadening out by including the Biological Engineering research. One of the major core areas of the conference program is life quality, due to the importance that Chemical and Biological Engineering plays in this area. “Integration of Life Sciences & Engineering” and “Sustainable Process-Product Development through Green Chemistry” are two of the leading themes with papers addressing such important issues. This is complemented with additional leading themes including “Advancing the Chemical and Biological Engineering Fundamentals”, “Multi-Scale and/or Multi-Disciplinary Approach to Process-Product Innovation”, “Systematic Methods and Tools for Managing the Complexity”, and “Educating Chemical and Biological Engineers for Coming Challenges” which define the extended abstracts arrangements along this book. A total of 516 extended abstracts are included in the book, consisting of 7 invited lecturers, 15 keynote, 105 short oral presentations given in 5 parallel sessions, along with 6 slots for viewing 389 poster presentations. Full papers are jointly included in the companion Proceedings in CD-ROM. All papers have been reviewed and we are grateful to the members of scientific and organizing committees for their evaluations. It was an intensive task since 610 submitted abstracts from 45 countries were received. It has been an honor for us to contribute to setting up CHEMPOR 2008 during almost two years. We wish to thank the authors who have contributed to yield a high scientific standard to the program. We are thankful to the sponsors who have contributed decisively to this event. We also extend our gratefulness to all those who, through their dedicated efforts, have assisted us in this task. On behalf of the Scientific and Organizing Committees we wish you that together with an interesting reading, the scientific program and the social moments organized will be memorable for all.Fundação para a Ciência e a Tecnologia (FCT

Animal Feed Science and Nutrition

Animal food production faces many challenges including shortage of high-quality feed ingredients, contribution of greenhouse gases and pollutants to the environment, development of antimicrobial resistance, food safety, and animal health and welfare. This examines these issues over three sections. The first section is the introduction. The second section provides insights into optimization of pasture utilization employing different supplements and feed additives to maximize beef cattle production, use of insect meal as a promising protein feed ingredient, and ruminal microbiome manipulation to improve ruminal fermentation efficiency. The third section discusses accurate estimation of enteric methane emission factors, reduction of enteric methane emissions by means of feeding management and antimethanogenic compounds, and different heavy metal pollution by poultry wastes and associated health hazards

Impact of Osmotic Stressors on the Metabolic Activity of Methylocystis sp. Strain SC2

Proteobacterial methane-oxidizing bacteria, or methanotrophs, have the unique ability to grow on methane as their sole source of carbon and energy. Among these, Methylocystis spp. belong to the family Methylocystaceae within the Alphaproteobacteria. Their key enzyme is the particulate methane monooxygenase (pMMO), which oxidizes methane to methanol. Methylocystis spp. are among the ecologically most relevant methanotroph populations in terrestrial environments and are widely distributed in diverse habitats. In consequence, Methylocystis spp. require a range of physiological capabilities that allow them to respond and acclimatize to fluctuations in abiotic and biotic factors in their native environment. However, to date there still exist major gaps in our knowledge of their metabolic potential, in particular with regard to their ability to acclimatize to environmental change and to cope with abiotic stress. In my first project, we used a recently developed proteome workflow to elucidate the cellular mechanisms underlying the acclimatization of Methylocystis sp. strain SC2 to high NH4+ load (added as NH4Cl). Relative to 1 mM NH4+, high (50 mM and 75 mM) NH4+ load under CH4-replete conditions significantly increased the lag phase duration required for proteome adjustment, while the addition of 100 mM NH4+ completely inhibited growth of strain SC2. The number of differentially regulated proteins was highly significantly correlated to the increase in NH4+ load. The cellular responses involved the significant upregulation of stress-responsive proteins, the K+ “salt-in” strategy, the synthesis of compatible solutes (glutamate and proline), and the glutathione metabolism. The apparent Km value for CH4 oxidation significantly increased with the NH4+ load. This observation was indicative of an increased pMMO-based oxidation of NH3 to toxic hydroxylamine. In consequence, the detoxifying activity of hydroxlyamine oxidoreductase (HAO) increased with the NH4+ concentration and led to a significant accumulation of NO2− and, with delay, N2O. Significant production of N2O occurred only after the oxygen concentration had dropped to low or unmeasurable levels. Thus, high NH4+ load had a dual effect on the activity of strain SC2, with one being general phenomenon of ionic-osmotic stress and the other being the competitive inhibition effect of NH3 on pMMO-based methane oxidation. Although strain SC2 precisely rebalanced enzymes and osmolyte composition in response to the increase in NH4+ load, the need to simultaneously combat both ionic-osmotic stress and the toxic effects of hydroxylamine may be the reason why its acclimatization capacity is limited to 75 mM NH4+. Starting point of my second project was the knowledge that the growth of strain SC2 is completely inhibited at medium concentrations  1.5% NaCl. Sodium chloride is an important ionic-osmotic stressor in bulk and rhizosphere soils. We therefore tested various amino acids and other osmolytes for their potential to act as a compatible solute or osmoprotectant under otherwise inhibitory NaCl conditions. The addition of 10 mM asparagine to the growth medium had the greatest stress relief effect under severe salinity (1.5% NaCl), leading to a partial growth recovery of strain SC2. The analysis of the exo-metabolome revealed that asparagine was taken up quantitatively by strain SC2. This resulted in an intracellular concentration of 264 ± 57 mM asparagine. Under severe salinity (1.5% NaCl), the uptake of asparagine induced major proteome rearrangements related to the KEGG level 2 categories energy metabolism, amino acid metabolism, and cell growth and death. In particular, various proteins involved in cell division and peptidoglycan synthesis showed a positive expression response. The incorporation of asparagine-derived 13C-carbon into nearly all amino acids indicated that asparagine acted as a source for cell biomass under severe salinity (1.5% NaCl), with glutamate being a major hub between central carbon and amino acid pathways