Carbohydrate Biotechnology Protocols

We are in a phase of the evolution of biotechnology in which the true and potential commercial importance of carbohydrates is becoming appre- ated more fully. Progress in providing hard facts to establish the commercial value ofpolysaccharides and oligosaccharides is limited, as always, by lack of funding and by a relative shortage of skilled practitioners in the production and analysis of those materials. Carbohydrate science has a reputation, not unmerited, for technical difficulty owing to the structural similarity of the many monosaccharide monomers and the potential, and real, complexity of oligosaccharides and polysaccharides, particularly heterosaccharides conta- ing many different monomers. Modem analytical and synthetic methods, in many cases using enzyme technology, are beginning to allow this complexity to be unraveled. Carbohydrate Biotechnology Protocols is aimed at those newcomers who have an interest in the production and use of carbohydrate materials, but have shied away from involvement for lack of detailed descriptions of appropriate methods, including the type of practical hints that may be provided by those skilled in those methods, but that are rarely described in research papers. The majority of the contributions to this book conform to the established format of the Methods in Biotechnology series. They begin with the theoretical and c- mercial background to the method or group of methods, provide a list of the reagents and equipment required for the procedure, then give a detailed st- by-step description of how to carry out the protocol.

This second edition provides new and updated tools for studying protein-carbohydrate interactions ranging from traditional biochemical methods to state-of-the-art techniques. This book focuses on four different research themes detailing methods for screening and quantifying CAZyme activity, investigating the interactions between proteins, carbohydrate ligands, methods for the visualization of carbohydrates, protein-carbohydrate complexes, structural and “omic” approaches for studying systems of CAZymes. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Carbohydrate- Protein Interactions: Methods and Protocols, Second Edition aims to be comprehensive guide for researchers in the field.

These volumes of Methods in Enzymology contain the protocols that made up the on-line Methods Navigator. Our philosophy when we selected the protocols to include in the Navigator was that they should be for techniques useful in any biomedical laboratory, regardless of the system the lab studies. Each protocol was written by researchers who use the technique routinely, and in many cases by the people who actually developed the procedure in the first place. The protocols are very detailed and contain recipes for the necessary buffers and reagents, as well as flow-charts outlining the steps involved. Many of the chapters have accompanying videos demonstrating key parts of the procedures. The volumes are broken into distinct areas: DNA methods; Cell-based methods; lipid, carbohydrate and miscellaneous methods; RNA methods; protein methods. Our goal is that these protocols will be useful for everyone in the lab, from undergraduates and rotation students to seasoned post-doctoral fellows. We hope that these volumes will become dog-eared and well-worn in your laboratory, either physically or electronically.

"This volume is a wide-ranging tool for studying protein-carbohydrate interactions that extend from traditional biochemical methods to state-of-the-art techniques. This book focuses on four different research themes: Part I describes methods for screening and quantifying CAZyme activity; Part II contains methods for investigating the interactions between proteins and carbohydrate ligands; Part III discusses methods for the visualization of carbohydrates and protein-carbohydrate complexes; and Part IV focuses on structural and "omic" approaches for studying systems of CAZymes. Written in the highly successful Methods in Molecular Biology series format, 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. Cutting-edge and thorough, Protein-Carbohydrate Interactions: Methods and Protocols is a valuable resource to the glycomics research community. In this continuously advancing field, the methods in this book highlight the biology of glycomics, thus driving biotechnological innovation and solutions for human health and sustainable resources within the emerging green community. ."--Prové de l'editor.

Glycobiology involves studies of complex carbohydrates and posttrans- tional modifications of proteins, and has become an important interdiscip- nary field encompassing chemistry, biochemistry, biology, physiology, and pathology. Although initial research was directed toward elucidation of the different carbohydrate structures and the enzymes synthesizing them, the field has now moved toward identifying the functions of carbohydrates. The pro- cols described in Glycobiology Protocols form a solid basis for investigations of glycan functions in health and disease. The cloning of many of the genes participating in glycosylation processes has helped to enhance our knowledge of how glycosylation is controlled, but has also added another dimension of complexity to the great heterogeneous variety of the structures of the oligos- charides of glycoproteins, proteoglycans, and glycolipids. A family of similar enzyme proteins exists for each glycosylation step. Glycosyltransferases are extremely specific for both the nucleotide sugar donor and the acceptor s- strate, but many other factors control sugar transfer, including the locali- tion and topology of enzymes, cofactors, possible chaperone proteins, and the availability of sugar acceptor substrates. The analysis of the intracellular organization of glycosylation and of the factors controlling the activities of the participating enzymes in the cell are important areas that need more research efforts. Another challenge for future research is to understand the glycodynamics of a cell, that is, how the cell responds to stimuli leading to biological and pathological changes in terms of alterations in glycosylation, and how this affects the biology of the cell.

The second edition of this book constitutes a comprehensive manual of new techniques for setting up mammalian cell lines for production of biopharmaceuticals, and for optimizing critical parameters for cell culture considering the whole cascade from lab to final production. The chapters are written by world-renowned experts and the volume’s five parts reflect the processes required for different stages of production. This book is a compendium of techniques for scientists in both industrial and research laboratories that use mammalian cells for biotechnology purposes.

Since its inception in 1945, this serial has provided critical articles written by research specialists that integrate industrial, analytical, and technological aspects of biochemistry, organic chemistry, and instrumentation methodology in the study of carbohydrates. Features contributions from leading authorities and industry experts Informs and updates on all the latest developments in the field

Two of the recent books in the Methods in Molecular Biology series, Yeast Protocols and Pichia Protocols, have been narrowly focused on yeasts and, in the latter case, particular species of yeasts. Food Microbiology Pro- cols, of necessity, covers a very wide range of microorganisms. Our book treats four categories of microorganisms affecting foods: (1) Spoilage organisms; (2) pathogens; (3) microorganisms in fermented foods; and (4) microorganisms p- ducing metabolites that affect the flavor or nutritive value of foods. Detailed information is given on each of these categories. There are several chapters devoted to the microorganisms associated with fermented foods: these are of increasing importance in food microbiology, and include one bacteriophage that kills the lactic acid bacteria involved in the manufacture of different foods—cottage cheese, yogurt, sauerkraut, and many others. The other nine chapters give procedures for the maintenance of lactic acid bacteria, the isolation of plasmid and genomic DNA from species of Lac- bacillus, determination of the proteolytic activity of lactic acid bacteria, det- mination of bacteriocins, and other important topics.