This volume provides an essential update on fundamental issues, current and new applications, as well as practical protocols to explore the extensive applications of lipases and the potential application of phospholipases. After an overview, the book delves into activity screening and expression, optimization of the biocatalyst production and performances, and applications of lipases, phospholipases, and esterases. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Lipases and Phospholipases: Methods and Protocols, Second Editionserves as an updated reference book for the large scientific community, both seasoned and novice, working with lipases, phospholipases, and related enzymes.

The lipases and phospholipases represent a diverse group of enzymes that are expressed in animals, plants, fungi, and bacteria. Their ubiquitous distribution among all species is a testament to the essential roles played by these enzymes in lipid storage, mobilization and metabolism, membrane homeostasis and remodeling, endocrine and immune functions, and signal tra- duction. In humans, lipases and phospholipases are also thought to contribute to complex diseases, such as atherosclerosis, obesity, arthritis, and cancer, as well as to single gene defects, such as Wolman's disease and Type I hyperlipoproteinemia. Enzymatically, the lipases are unique, since they hydrolyze substrates that are either insoluble, or only partly soluble, in aq- ous solvents; thus, enzyme catalysis takes place at a lipid-water interface. The interface comprises at least two, and often more, discrete bulk and s- face phases, in which the enzyme, substrate, and products oflipolysis disperse among these phases based on their physical properties. Furthermore, the d- tribution of these components changes continuously as lipolysis proceeds. Thus, the lipases and phospholipases are fundamentally different from any other enzyme because of the physical complexity of the environment in which catalysis occurs.

A collection of papers that comprehensively describe the major areas of research on lipid metabolism of plants. State-of-the-art knowledge about research on fatty acid and glycerolipid biosynthesis, isoprenoid metabolism, membrane structure and organization, lipid oxidation and degradation, lipids as intracellular and extracellular messengers, lipids and environment, oil seeds and gene technology is reviewed. The different topics covered show that modern tools of plant cellular and molecular biology, as well as molecular genetics, have been recently used to characterize several key enzymes of plant lipid metabolism (in particular, desaturases, thioesterases, fatty acid synthetase) and to isolate corresponding cDNAs and genomic clones, allowing the use of genetic engineering methods to modify the composition of membranes or storage lipids. These findings open fascinating perspectives, both for establishing the roles of lipids in membrane function and intracellular signalling and for adapting the composition of seed oil to the industrial needs. This book will be a good reference source for research scientists, advanced students and industrialists wishing to follow the considerable progress made in recent years on plant lipid metabolism and to envision the new opportunities offered by genetic engineering for the development of novel oil seeds.

Enzymatic methods of lipid modification, particularly of fats and oils, have developed rapidly since the 1980s. In parallel to the rapid progress in research a wide range of applications have emerged, e.g. in the food industry. The book is written by leading experts in the field and reflects the state-of-the-art of enzymatic lipid modification. It provides the reader with guidelines how to select suitable enzymes and how to apply them efficiently. Applications of lipases and phospholipases, lipoxygenases and P450-monooxygenases and the use of whole-cell systems in lipid modification are described. Cloning, expression and mutagenesis as well as attempts to understand the molecular basis of specificity and stereoselectivity are outlined. In addition engineering aspects and the choice of solvent systems are addressed.

“Infogest” (Improving Health Properties of Food by Sharing our Knowledge on the Digestive Process) is an EU COST action/network in the domain of Food and Agriculture that will last for 4 years from April 4, 2011. Infogest aims at building an open international network of institutes undertaking multidisciplinary basic research on food digestion gathering scientists from different origins (food scientists, gut physiologists, nutritionists...). The network gathers 70 partners from academia, corresponding to a total of 29 countries. The three main scientific goals are: Identify the beneficial food components released in the gut during digestion; Support the effect of beneficial food components on human health; Promote harmonization of currently used digestion models Infogest meetings highlighted the need for a publication that would provide researchers with an insight into the advantages and disadvantages associated with the use of respective in vitro and ex vivo assays to evaluate the effects of foods and food bioactives on health. Such assays are particularly important in situations where a large number of foods/bioactives need to be screened rapidly and in a cost effective manner in order to ultimately identify lead foods/bioactives that can be the subject of in vivo assays. The book is an asset to researchers wishing to study the health benefits of their foods and food bioactives of interest and highlights which in vitro/ex vivo assays are of greatest relevance to their goals, what sort of outputs/data can be generated and, as noted above, highlight the strengths and weaknesses of the various assays. It is also an important resource for undergraduate students in the ‘food and health’ arena.

Interfacial Phenomena examines the fundamental properties of various liquid interfaces. This book discusses the physics of surfaces; electrostatic and electrokinetic phenomena; and adsorption at liquid interfaces. The properties of monolayers; reactions at liquid surfaces; diffusion through interfaces; and disperse systems and adhesion are also deliberated. Other topics include the vapor pressures over curved surfaces; electrical capacity of the double layer; applications of electrophoresis; and thermodynamics of adsorption and desorption. The experimental methods of spreading films at the oil-water interface; penetration into monolayers; experiments on dynamic systems; and spontaneous emulsification are likewise covered in this text. This book is beneficial to chemical engineers and students concerned with interfacial phenomena.

“Microbial Enzymes: Roles and applications in industry” offers an essential update on the field of microbial biotechnology, and presents the latest information on a range of microbial enzymes such as fructosyltransferase, laccases, amylases, lipase, and cholesterol oxidase, as well as their potential applications in various industries. Production and optimisation technologies for several industrially relevant microbial enzymes are also addressed. In recent years, genetic engineering has opened up new possibilities for redesigning microbial enzymes that are useful in multiple industries, an aspect that the book explores. In addition, it demonstrates how some of the emerging issues in the fields of agriculture, environment and human health can be resolved with the aid of green technologies based on microbial enzymes. The topics covered here will not only provide a better understanding of the commercial applications of microbial enzymes, but also outline futuristic approaches to use microbial enzymes as driver of industrial sustainability. Lastly, the book is intended to provide readers with an overview of recent applications of microbial enzymes in various industrial sectors, and to pique researchers’ interest in the development of novel microbial enzyme technologies to meet the changing needs of industry.

Lipases and Phospholipases are key control elements in mammalian metabolism. They share many common features that set them apart from other metabolic enzyme classes, most importantly their association with biological membranes. Their potential as drug targets for the treatment of metabolic diseases is widely recognized, and the first lipase inhibitor drugs have been successfully introduced. Providing drug developers with a firm foundation for lipase-centered drug design, the editors of this volume have assembled experts from different scientific disciplines to create a comprehensive handbook for all pharmaceutical chemists, biochemists and physiologists working with lipases. The authors examine fundamental aspects of lipase function in vitro and in vivo, explaining how this knowledge may be used to develop lipase assays. They also treat the physiological roles of lipases in normal and disordered metabolism, as well as strategies to target lipases for the treatment of diabetes, obesity and related disorders. Additional topics include the application of phospholipases for liposome-based drug delivery and their use as diagnostic tools.

The endothelium, a monolayer of endothelial cells, constitutes the inner cellular lining of the blood vessels (arteries, veins and capillaries) and the lymphatic system, and therefore is in direct contact with the blood/lymph and the circulating cells. The endothelium is a major player in the control of blood fluidity, platelet aggregation and vascular tone, a major actor in the regulation of immunology, inflammation and angiogenesis, and an important metabolizing and an endocrine organ. Endothelial cells controls vascular tone, and thereby blood flow, by synthesizing and releasing relaxing and contracting factors such as nitric oxide, metabolites of arachidonic acid via the cyclooxygenases, lipoxygenases and cytochrome P450 pathways, various peptides (endothelin, urotensin, CNP, adrenomedullin, etc.), adenosine, purines, reactive oxygen species and so on. Additionally, endothelial ectoenzymes are required steps in the generation of vasoactive hormones such as angiotensin II. An endothelial dysfunction linked to an imbalance in the synthesis and/or the release of these various endothelial factors may explain the initiation of cardiovascular pathologies (from hypertension to atherosclerosis) or their development and perpetuation. Table of Contents: Introduction / Multiple Functions of the Endothelial Cells / Calcium Signaling in Vascular Cells and Cell-to-Cell Communications / Endothelium-Dependent Regulation of Vascular Tone / Conclusion / References