Today, activation endoproteolysis of secretory proteins is recognized as a fundamental biological mechanism of spatial and temporal regulation of protein activity as well as of diversification of protein functions. In Proprotein Convertases, experts in the field examine detailed methods involving proprotein convertases, the enzymes mediating this endoproteolysis, which reside within or cycle between the various compartments of the secretory pathway. Providing a timely assessment of impact of activation/inactivation endoproteolysis in the secretory pathway, the volume offers a broader perspective on the biochemistry of the PCSKs (proprotein convertases, subtilisin/kexin-type) by exploring structural and functional analogies with bacterial subtilisin and on the enzymology of endoproteolysis itself by describing the involvement in the process of non-PCSK-type such as cathepsin L. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their specific topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Meticulous and up-to-date, Proprotein Convertases represents an instructive and useful reference book for all scientists interested in endoproteolytic activation and/or inactivation of secretory proproteins through limited proteolysis, for experts in the field and newcomers to it as well.

The last fifteen years have witnessed the birth and maturation of many original methods and the development of protocols specific to single molecule measurements and their analysis, including techniques involving optical imaging, electron microscopy, optical and magnetic trapping, and developments in atomic force microscopy. In Single Molecule Enzymology: Methods and Protocols, experts in the field provide procedures which enable the extraction of detailed information about enzyme work cycles, their static and kinetic properties, and information about their location and activity within cells. The detailed volume offers practical advice on many aspects of single molecule enzymology and includes strategic overviews of interconnected methods involved in sample preparation, single molecule measurements, and data analysis. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters contain 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 up-to-date, Single Molecule Enzymology: Methods and Protocols is intended for use within the diverse community of molecular biologists, biochemists, and biophysicists studying enzymes in detail and can be used by researchers planning their first single molecule study or to aid more experienced researchers in further developing their existing studies.

Whether the pursuit is commercially motivated or purely academic, engineering a novel biological catalyst is an enticing challenge. High-resolution protein structure analysis allows for rational alteration of enzyme function, yet many useful enzyme variants are the product of well-designed selection schemes or screening strategies. Enzyme Engineering: Methods and Protocols provides guidance to investigators wishing to create enzyme variants with desired properties. This detailed volume covers such topics as a simple method for generating site-specific mutations within bacterial chromosomes. It also highlights the engineering of two difference types of rare-cutting endonucleases that show great potential in gene therapy applications: The newest development is the emergence of TAL effector nucleases or TALENs. Chapters describe newly developed technologies in sufficient detail so that each method can be practiced in a standard molecular biology laboratory. Written in the successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible Enzyme Engineering: Methods and Protocols will be valuable for scientists with a budding interest in protein engineering as well as veterans looking for new approaches to apply in established discovery programs.

In this thesis, Xiaoshi Wang investigates the function and mechanism of a newly discovered heme-thiolate peroxygenase, AaeAPO. This enzyme class comes from Agrocybe aegerita and is used in the conversion of inert hydrocarbons to alcohols. Xiaoshi's work focuses on an extracellular P450 enzyme which is not limited in its stability and lack of solubility and therefore is relevant for widespread industrial use. The author demonstrates that the peroxygenase catalyzes a wide range of reactions. In some cases the author even describes very difficult transformations in molecules that are highly inert. Her detailed investigations provide a mechanistic framework for how the peroxygenase catalyzes such a large number of reactions. A major highlight of this thesis is the identification of key short-lived intermediates in the catalytic cycle of the peroxygenase, using rapid kinetic and spectroscopic methods, as well as the elucidation of the thermodynamic properties of these high-energy intermediates. This work adds new insight into an important class of enzymes.

Tryptophan metabolism via kynurenine pathway plays a critical role in both health and a variety of human diseases. This book highlights the known associations between kynurenine pathway and various disease states, as well as examines the current status of drug development and clinical trials of compounds known to alter tryptophan metabolism. The research plays a critical role in molecular targeted therapies directed at altering the kynurenine pathway of tryptophan metabolism. The initial and rate-limiting step of tryptophan metabolism is mediated by one of two enzymes, tryptophan-2,3-dioxygenase (TDO; predominantly in the liver, but also in the brain) and indoleamine-2,3-dioxygenase (IDO; in a host of tissues in response to immune activation). Targeting the enzymes IDO and TDO, as well as other downstream effectors would therefore be likely to generate novel treatment options that would be helpful in a wide variety of clinical settings. This book provides a unique bridge between basic mechanistic understanding of the role of the kynurenine pathway with translational applications and clinical relevance. It will explore the indications that tryptophan metabolism is a potential biomarker of disease activity, can contribute to local and possibly systemic immune suppression in cancer, and is an attractive target for which a variety of inhibitors are readily available.

This excellent book covers wide-ranging topics in interdisciplinary microbiology, addressing various research aspects and highlighting advanced discoveries and innovations. It presents the fascinating topic of modern biotechnology, including agricultural microbiology, microalgae biotechnology, bio-energy, bioinformatics and metagenomics, environmental microbiology, enzyme technology and marine biology. It presents the most up-to-date areas of microbiology with an emphasis on shedding light on biotechnological advancements and integrating these interdisciplinary microbiology research topics into other biotechnology sub-disciplines. The book raises awareness of the industrial relevance of microbiology, which is key component of this unique collection. The topics include production of antioxidant-glutathione, enzyme-engineering methods, probiotic microbiology and features of microbial xylanases. It also covers some other remarkable aspects of microbiology, like potential health hazards in recreational water and fullerene nanocomposites, which are vital for biotechnological interventions. This book will be valuable resource for senior undergraduate and graduate students, researchers and other interested professionals or groups working in the interdisciplinary areas of microbiology and biotechnology.

Since the discovery of a collagen-degrading protease in the tadpole tail in 1962, matrix metalloproteinase research has led to the discovery of more than twenty distinct vertebrate MMPs, along with a variety of homologues from diverse organisms such as the sea urchin, plants, insects, and nematode worms. Fully updating and adding to the popular first edition, Matrix Metalloproteinase Protocols, Second Edition includes a series of state-of-the-art techniques provided by eminent experts in the field. Beginning with a brief overview of the MMP arena, from how these enzymes fit into the larger degradome to what occurs when their expression and function in the mouse is modulated, the volume continues with sections on the expression and purification of MMPs and TIMPs, the detection of MMPs and TIMPs at both the protein and mRNA level, and our ability to assay MMP and TIMP activities in a wide variety of circumstances. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, Matrix Metalloproteinase Protocols, Second Edition is an ideal source for many of the essential laboratory techniques for both novice and seasoned researchers alike collected in one convenient volume.

In this brief, the authors explore and review the current knowledge regarding the role of molybdenum in the evolution of biological systems and their interaction with biogeochemical cycles. Special emphasis is placed on biological nitrogen fixation and the nitrogen element cycle. The origin and evolution of molybdenum cofactor biosynthetic pathways as well as the evolutionary significance of molybdenum containing enzymes appearance is analyzed. Original data regarding nitrogen fixation pathways and related enzymes molecular evolution processes is presented. The trace element molybdenum is essential for nearly all organisms and forms the catalytic center of a large variety of enzymes such as nitrogenase, nitrate reductases, sulphite oxidase and xanthine oxidoreductases.

Yeasts are a versatile group of eukaryotic microorganisms, exhibiting heterogeneous nutritional profiles and an extraordinary ability to survive in a wide range of natural and man-associated ecosystems, including cold habitats. Cold-adapted yeasts inhabit numerous low-temperature environments where they are subjected to seasonal or permanent cold conditions. Hence, they have evolved a number of adaptation strategies with regard to growth and reproduction, metabolic activities, survival and protection. Due to their distinctive ability to thrive successfully at low and even subzero temperatures, cold-adapted yeasts are increasingly attracting attention in basic science and industry for their enormous biotechnological potential. This book presents our current understanding of the diversity and ecology of cold-adapted yeasts in worldwide cold ecosystems, their adaptation strategies, and their biotechnological significance. Special emphasis is placed on the exploitation of cold-adapted yeasts as a source of cold-active enzymes and biopolymers, as well as their benefits for food microbiology, bioremediation and biocontrol. Further, aspects of food biodeterioration are considered.

Alexander Todd, the 1957 Nobel laureate in chemistry is credited with the statement: "where there is life, there is phosphorus". Phosphorus chemical biology underlies most of life's reactions and processes, from the covalent bonds that hold RNA and DNA together, to the making and spending 75 kg of ATP every day, required to run almost all metabolic and mechanical events in cells. Authored by a renowned biochemist, The Chemical Biology of Phosphorus provides an in-depth, unifying chemical approach to the logic and reactivity of inorganic phosphate and its three major derivatives (anhydrides, mono- and diesters) throughout biology to examine why life depends on phosphorus. Covering the breadth of phosphorus chemistry in biology, this book is ideal for biochemistry students, postgraduates and researchers interested in the chemical logic of phosphate metabolites, energy generation, biopolymer accumulation and phosphoproteomics.

This book describes the complex processes involved in styrene degradation by microbes, including highly adaptive microorganisms, the various enzymes involved in styrene biodegradation, new styrene-catabolic routes, novel regulatory mechanisms, and the genes coding for styrene metabolizing enzymes. Numerous biotechnological applications are discussed, such as the development of sustainable eco-friendly technologies as well as the use of styrene degrading microorganisms and their enzymes as a rich resource for biotechnology.

Springer Handbook of Enzymes provides data on enzymes sufficiently well characterized. It offers concise and complete descriptions of some 5,000 enzymes and their application areas. Data sheets are arranged in their EC-Number sequence and the volumes themselves are arranged according to enzyme classes. This new, second edition reflects considerable progress in enzymology: many enzymes are newly classified or reclassified. Each entry is correlated with references and one or more source organisms. New datafields are created: application and engineering (for the properties of enzymes where the sequence has been changed). The total amount of material contained in the Handbook has more than doubled so that the complete second edition consists of 39 volumes as well as a Synonym Index. In addition, starting in 2009, all newly classified enzymes are treated in Supplement Volumes. Springer Handbook of Enzymes is an ideal source of information for researchers in biochemistry, biotechnology, organic and analytical chemistry, and food sciences, as well as for medicinal applications.

Springer Handbook of Enzymes provides data on enzymes sufficiently well characterized. It offers concise and complete descriptions of some 5,000 enzymes and their application areas. Data sheets are arranged in their EC-Number sequence and the volumes themselves are arranged according to enzyme classes. This new, second edition reflects considerable progress in enzymology: many enzymes are newly classified or reclassified. Each entry is correlated with references and one or more source organisms. New datafields are created: application and engineering (for the properties of enzymes where the sequence has been changed). The total amount of material contained in the Handbook has more than doubled so that the complete second edition consists of 39 volumes as well as a Synonym Index. In addition, starting in 2009, all newly classified enzymes are treated in Supplement Volumes. Springer Handbook of Enzymes is an ideal source of information for researchers in biochemistry, biotechnology, organic and analytical chemistry, and food sciences, as well as for medicinal applications.

The Springer Handbook of Enzymes provides concise data on some 5,000 enzymes sufficiently well characterized - and here is the second, updated edition. Their application in analytical, synthetic and biotechnology processes as well as in food industry, and for medicinal treatments is added. Data sheets are arranged in their EC-Number sequence. The new edition reflects considerable progress in enzymology: the total material has more than doubled, and the complete 2nd edition consists of 39 volumes plus Synonym Index. Starting in 2009, all newly classified enzymes are treated in Supplement Volumes.

The Springer Handbook of Enzymes provides concise data on some 5,000 enzymes sufficiently well characterized - and here is the second, updated edition. Their application in analytical, synthetic and biotechnology processes as well as in food industry, and for medicinal treatments is added. Data sheets are arranged in their EC-Number sequence. The new edition reflects considerable progress in enzymology: the total material has more than doubled, and the complete 2nd edition consists of 39 volumes plus Synonym Index. Starting in 2009, all newly classified enzymes are treated in Supplement Volumes.

The Springer Handbook of Enzymes provides concise data on some 5,000 enzymes sufficiently well characterized - and here is the second, updated edition. Their application in analytical, synthetic and biotechnology processes as well as in food industry, and for medicinal treatments is added. Data sheets are arranged in their EC-Number sequence. The new edition reflects considerable progress in enzymology: the total material has more than doubled, and the complete 2nd edition consists of 39 volumes plus Synonym Index. Starting in 2009, all newly classified enzymes are treated in Supplement Volumes.

The Springer Handbook of Enzymes provides concise data on some 5,000 enzymes sufficiently well characterized - and here is the second, updated edition. Their application in analytical, synthetic and biotechnology processes as well as in food industry, and for medicinal treatments is added. Data sheets are arranged in their EC-Number sequence. The new edition reflects considerable progress in enzymology: the total material has more than doubled, and the complete 2nd edition consists of 39 volumes plus Synonym Index. Starting in 2009, all newly classified enzymes are treated in Supplement Volumes.

Cutting edge reviews by leading researchers illuminate key aspects of DNA repair in mammalian systems and its relationship to human genetic disease and cancer. Major topics include UV and X-Ray repair, repair of chemical damage, recombinational repair, mismatch repair, transcription-repair coupling, and the role of DNA repair in disease prevention. Extensive up-to-date references and rigorous peer-review of each chapter make this volume definitive and bring it to the active frontiers of research.

Accessible and comprehensive, this book describes the universal cellular nature of living organisms and is an indispensable tool for anyone in the sciences who wishes to get a quick overview of molecular biology. Individual chapters cover nucleic acids and proteins, genetic code and protein synthesis, the fidelity of transferring genetic information to the next generations, and the regulation of various processes inside the cells. Special attention is paid to new areas rising from modern DNA sequencing technologies which transform biology. The book also touches on developing areas, such as cures for cancer and CRISPR, which are important for medicine and the future of humankind.

Gamma-Glutamyl Transpeptidases ( -GTs) are members of the N-terminal nucleophile hydrolase superfamily, enzymes that cleave the -glutamyl amide bond of glutathione to liberate cysteinylglycine. The released -glutamyl group can be transferred to water (hydrolysis) or to amino acids or short peptides (transpeptidation). -GT plays a key role in the gamma glutamyl cycle by regulating the cellular levels of the antioxidant glutathione, hence it is a critical enzyme in maintaining cellular redox homeostasis. -GT is upregulated during inflammation and in several human tumors, and it is involved in many physiological disorders related to oxidative stress, such as Parkinson's disease and diabetes. Furthermore, this enzyme is used as a marker of liver disease and cancer. This book covers current knowledge about the structure-function relationship of -GTs and gives information about applications of -GTs in different fields ranging from clinical biochemistry to biotechnology and biomedicine.

This textbook is designed for students of biology, molecular biology, ecology,medicine, agriculture, forestry and other professions where the knowledge of organic chemistry plays an important role. The work may also be of interest to non-professionals, as well as to teachers in high schools. The book consists of 13 chapters that cover the essentials of organic chemistry, including - basic principles of structure and constitution of organic compounds, - the elements of the nomenclature, - the concepts of the nature of chemical bond, - introductions in NMR and IR spectroscopy, - the concepts and main classes of the organic reaction mechanisms, - reactions and properties of common classes or organic compounds, - and the introduction to the chemistry of the natural organic products followed by basic principles of the reactions in living cells. This second edition includes revisions and suggestions made by the readers of the first edition and the author's colleagues. In addition, it includes substantial changes compared to the first edition. The chapter on Cycloaddition has been completed by including the other pericyclic reactions (sigmatropic rearrangements, electrocyclic reactions). The chapter on Organic Natural Products has been extended to include new section covering the principles of organic synthesis. New chapter "Organic Supramolecular and Supermolecular Structures" is added. This chapter covers the basic knowledge about the molecular recognition, supramolecular structures, and the mechanisms of the enzyme catalyzed reactions.

The second international symposium on Pectins and Pectinases was organized by Wageningen University and Research Centre and held in Rotterdam, May 6-10, 2001. This successful meeting was attended by around 130 participants from more than 20 countries representing almost all of the groups and industries working woridwide on pectins and pectinases. Following the first meeting on this subject held in December 1995, the symposium defInitely forms a platform for researchers and industries working in the fIeld, all within their own discipline and expertise. The symposium demanded a written account and this book is the resuit of that. It contains aIl keynote lectures and other oral presentations and provides an update about the current research. SignifIcant progress has been made in the last 5 years. This book provides an up-to-date insight into the research on pectin and pectic enzymes involved in their biosynthesis, degradation, modifIcation or utilization. The progress in the elucidation of the chemical structure of pectin and mode of action and 3-D structure of the pectin degrading enzymes allows us to identify and influence the functionality of pectins and pectic enzymes, both in vitra after isolation as weIl in the plants themselves (in planta). Other contributions deal with new applications of both pectin and pectin-degrading enzymes, while more and more attention is paid to health and nutritional aspects ofpectins. The book provides a 'state of the art' account for both beginners and experienced researchers of almost all disciplines of pectin research.

Enzyme Kinetics and Mechanisms takes the reader through the experimental techniques and the logic by which the mechanisms of enzyme-catalyzed reactions can be elucidated by the results of steady-state kinetics and related experiments. It is meant to make these investigations both satisfying and effective. In distinction to other available descriptions, the descriptions in enzyme Kinetics and Mechanisms are limited to more commonly utilized and useful models and techniques. The logic relating the chemical models to the mathematical models and the logic of relating the mathematical models to data is presented in rather concise text, figures and equations. The development of mathematical models from chemical models is done by a unique algorithm that is both simple and quick, and the same concept are utilized to develop models for the effects of a variety of reaction conditions on the initial velocity. In addition, the various relationships of data, mathematical models and the chemical models is illustrated with examples from the scientific literature.

Enzyme Kinetics and Mechanisms is intended for research workers, graduate students, post doctoral associates, and faculty in biochemistry and molecular biology who are interested in the techniques and logic by which mechanisms of enzymes-catalyzed reactions can be elucidated by investigation of steady-state kinetic experiments.