This book delves into the field of immobilizing biologically active and non-active molecules. It discusses the designing strategy of immobilization and the current state-of-the-art applications for advancing biomedical, agricultural, environmental and industrial practices. It focuses on aspects ranging from fundamental principles to current technological advances at multi-scale levels (macro, micro, and nano) which are suitable for cell, enzyme, and nano-catalyst based applications. Written by experts from across the globe, the contents deal with illustrated examples of molecular and cellular interactions with materials/scaffolds and discussions on factors that can affect the functionality and yield of the process. With its discussions on material science, design of delivery vehicles, separation science, additive manufacturing, agriculture and environmental science, this book will be a useful reference for researchers across multiple disciplines.

This book of protocols is devoted to the yellow coenzymes derived from riboflavin or vitamin B2 and to the hundreds of enzymes whose functionality depends on them, and represents a compendium of techniques for working with flavoproteins or with the wide spectrum of functions that flavoproteins can drive in the cells. Starting with Rf production in microorganisms and the chemical, optical, and redox properties of these fascinating molecules and moving along to the variety and the peculiarity of some single flavoenzymes, the volume explores the complexity of functions and distribution of these molecules in the cell. 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 practical, Flavin and Flavoproteins: Methods and Protocols serves as an ideal guide for protein chemists interested in purifying and characterizing flavoproteins, as well as microbiologists, physiologists, and clinicians, who wish to further study problems connected with flavoproteins.

This 2nd edition of the book on DNA methyltransferases has been comprehensively updated to reflect many novel research findings regarding the structure, function, and technology of these enzymes that have emerged over the past 6 years. Like the previous edition, this 2nd edition explains the biochemical properties of DNA methyltransferases, describing their structures, mechanisms and biological roles in bacteria, humans and plants. It also discusses the biological processes of reading DNA methylation and the mechanisms of DNA demethylation. This volume highlights the newest findings on DNA methyltransferase inhibitors and their use in cancer therapy as well as the latest epigenome editing systems based on these enzymes. Overall, this 2nd edition comprehensively summarizes the current state of research in the field of DNA methylation and DNA methyltransferase and is essential reading for early career and advanced researchers in this exciting field.

This book is a translation of Emil Fischer's autobiography published in 1922 by Verlag von Julius Springer, Berlin, Germany. It is the first translation of this work into English, guiding the reader through the life of a man who was one of the greatest chemists of all time. Emil Fischer published very important papers on sugars, purines, and peptides. His proof of the stereo chemistry of glucose remains one of the great intellectual and practical achievements of science. The book is of great benefit to the current and future generations of chemists, giving them the chance to get to know Emil Fischer's life story.

The enzymology of milk and other products is of enormous significance for the production and quality of almost every dairy product. Milk itself is a complex biological fluid that contains a wide range of enzymes with diverse activities, some of which have identifiable functions while others are present as an accidental consequence of the mechanism of milk secretion. Over time milk enzymology has become an incredibly essential component of milk and other dairy product production, and with advancing technology and processing techniques, its importance is at its peak. Dairy Enzymology presents an expansive overview of the enzymology of milk and other dairy products, focusing on the use of indigenous and endogenous enzymes in milk and exogenous enzymes in cheese processing. A full section is dedicated to the enzymology of bovine milk, focusing on the main families of indigenous enzymes as well as their potential significance in the mammary gland plus the technological significance for the properties of dairy products. Implications for the manufacture and ripening of cheese plus the use of enzymes such as alkaline phosphatase for measuring heat treatment in milk are explored in full, and the role of milk protease plasmin and other indigenous enzymes in the age-gelation is focused on. Further sections focus on enzymes found in raw milk and enzymes deliberately added for manufacture or modification of properties and the manufacture of food ingredients from dairy-derived ingredients. The key bacterial families are discussed in depth as well as their known contributions to the quality of dairy products. With its comprehensive scope and fully up-to-date coverage of dairy product enzymology, this text is a singular source for researchers looking to understand this essential dairy processing aspect.

The development of agents capable of cleaving RNA and DNA has attracted considerable attention from researchers in the last few years, because of the immediate and very important applications they can find in the emerging fields of biotechnology and pharmacology. There are essentially two classes of these agents - nucleases that occur naturally inside cells and synthetically produced artificial nucleases. The first class includes protein enzyme nucle ases and catalytic RNA structured ribozymes that perform cleavage of the phosphodiester bonds in nucleic acids according to a hydrolytic pathway in the course of different biochemical processes in the cell. A different pathway is used by some antibiotics which cleave DNA via redox-based mechanisms resulting in oxidative damage of nucleotide units and breakage of the DNA backbone. The above molecules are indispensable tools for manipulating nucleic acids and processing RNA; DNA-cleaving antibiotics and cytotoxic ribonucleases have demonstrated utility as chemotherapeutic agents. The second class, artificial nucleases, are rationally designed to imitate the active centers of natural enzymes by simple structures possessing minimal sets of the most important characteristics that are essential for catalysis. A dif ferent approach, in vitro selection, was also used to create artificial RNA and DNA enzymes capable of cleaving RNA. Being less efficient and specific as compared to the natural enzymes, the primitive mimics are smaller and robust and can function in a broad range of conditions."

Ribonucleic acid (RNA) is a macromolecule that plays a central role in cell physiology: RNA molecules act as intermediates between the deoxyribonucleic acid (DNA), where genetic information is stored, and proteins, which perform the necessary functions within the cell. Traditionally, the structural and functional properties of RNA are closely linked to gene expression. However, RNA-based enzymes, called ribozymes, are also involved in catalysis and small RNAs regulate key cellular processes, such as cell growth, division, differentiation, aging and death. RNA is a sensitive macromolecule that can be easily damaged by environmental conditions (ultraviolet radiation, oxidative stress) and biological factors (ribonucleases, ribotoxins, CRISPR-Cas systems). Therefore, cells have developed mechanisms to protect and/or repair RNA molecules. This book presents an overview of the biology of RNA damage, protection and repair in prokaryotes and eukaryotes. Individual chapters cover the expression regulation, enzymology and physiological role of such systems, and link them to important human diseases such as cancer and degenerative diseases.

This book introduces the new concept of "nanozyme", which refers to nanomaterials with intrinsic enzymatic activity, rather than nanomaterials with biological enzymes incorporated on the surface. The book presents the cutting-edge advances in nanozyme, with emphasis on state-of-the-art applications in many important fields, such as in the biomedical fields and for environmental protection. The nanozyme is a totally new type of artificial enzyme and exhibits huge advantages over natural enzymes, including greater stability, low cost, versatility, simplicity, and suitability for industry. It is of interest to university researchers, R&D engineers, as well as graduate students in nanoscience and technology, and biology wishing to learn the core principles, methods, and the corresponding applications of "nanozyme".

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."

A modern approach to enzyme kinetics and its applications

As catalysts for the majority of metabolic and biochemical reactions in the body, enzymes are important drug targets as well as useful synthetic catalysts. Enzyme kinetics is the study of the speed of an enzyme-catalyzed reaction and provides useful knowledge that aids in the design of enzyme-based processes. A. G. Marangoni’s Enzyme Kinetics: A Modern Approach provides a practical, how-to guide for students, technicians, and nonspecialists to evaluate enzyme kinetics, using common software packages to perform easy enzymatic analyses.

The treatment of enzyme kinetics in this book is radically different from the way the topic is traditionally covered. Marangoni stresses an understanding of how researchers arrive at models, what the models’ limitations are, and how they can be used in practical ways to analyze enzyme kinetic data. With the advent of computers, linear transformations of models have become unnecessary–Enzyme Kinetics does away with all linear transformations of enzyme kinetic models, advancing the use of nonlinear regression techniques. Marangoni develops new ways to carry out analyses of enzyme kinetic data, particularly in the study of pH effects on catalytic activity and multisubstrate enzymes. Other topics addressed include:

• Tools and techniques of kinetic analysis
• Reversible and irreversible enzyme inhibition
• Multisite and cooperative enzymes
• Immobilized and interfacial enzymes
• Characterization of enzyme stability

Enzyme Kinetics is a handy, innovative resource for practicing researchers in the chemical, pharmaceutical, and food science industries.

Beginning with the Escherichia coli ? protein, or bacterial DNA topoisomerase I, an ever-increasing number of enzymes have been identified that catalyze changes in the linkage of DNA strands. DNA topoisomerases are ubiquitous in nature and have been shown to play critical roles in most p- cesses involving DNA, including DNA replication, transcription, and rec- bination. These enzymes further constitute the cellular targets of a number of clinically important antibacterial and anticancer agents. Thus, further studies of DNA topology and DNA topoisomerases are critical to advance our und- standing of the basic biological processes required for cell cycle progression, cell division, genomic stability, and development. In addition, these studies will continue to provide critical insights into the cytotoxic action of drugs that target DNA topoisomerases. Such mechanistic studies have already played an important role in the development and clinical application of antimicrobial and chemotherapeutic agents. The two volumes of DNA Topoisomerase Protocols are designed to help new and established researchers investigate all aspects of DNA topology and the function of these enzymes. The chapters are written by prominent investigators in the field and provide detailed background information and st- by-step experimental protocols. The topics covered in Part I: DNA Topology and Enzymes, range from detailed methods to analyze various aspects of DNA structure, from linking number, knotting/unknotting, site-specific recombi- tion, and decatenation to the overexpression and purification of bacterial and eukaryotic DNA topoisomerases from a variety of cell systems and tissues.

This book presents an authoritative review of the most significant findings about all the epigenetic targets (writers, readers, and erasers) and their implication in physiology and pathology. The book also covers the design, synthesis and biological validation of epigenetic chemical modulators, which can be useful as novel chemotherapeutic agents. Particular attention is given to the chemical mechanisms of action of these molecules and to the drug discovery prose which allows their identification. This book will appeal to students who want to know the extensive progresses made by epigenetics (targets and modulators) in the last years from the beginning, and to specialized scientists who need an instrument to quickly search and check historical and/or updated notices about epigenetics.

This essential volume comprehensively discusses redox-active therapeutics, focusing particularly on their molecular design, mechanistic, pharmacological and medicinal aspects. The first section of the book describes the basic aspects of the chemistry and biology of redox-active drugs and includes a brief overview of the redox-based pathways involved in cancer and the medical aspects of redox-active drugs, assuming little in the way of prior knowledge. Subsequent sections and chapters describe more specialized aspects of central nervous system injuries, neurodegenerative diseases, pain, radiation injury and radioprotection (such as of brain, lungs, head and neck and erectile function) and neglected diseases (e.g., leishmaniasis). It encompasses several major classes of redox-active experimental therapeutics, which include porphyrins, salens, nitrones, and most notably metal-containing (e.g., Mn, Fe, Cu, Zn, Sb) drugs as either single compounds or formulations with nanomaterials and quantum dots. Numerous illustrations, tables and figures enhance and complement the text; extensive references to relevant literature are also included. Redox-Active Therapeutics is an invaluable addition to Springer's Oxidative Stress in Applied Basic Research and Clinical Practice series. It is essential reading for researchers, clinicians and graduate students interested in understanding and exploring the Redoxome-the organism redox network-as an emerging frontier in drug design, redox biology and medicine.

This book provides a comprehensive, organized, and concise overview of xenobiotic metabolic enzymes and their health implications. The subjects addressed are broad in scope with an emphasis on recent advances in research on biochemical and biomedical aspects of these enzymes. The xenobiotics discussed include not just drugs, but also food, smoke, and other environmental chemicals. The subjects covered in this work include: metabolic enzymes and their catalyzed reactions, reactive intermediates generated from metabolic activation, oxidative stress mediated by electrophilic reactive intermediates, bioactivation - mediated cellular and functional damages, activation of Nrf2 - ARE pathway, genetic variations affecting metabolic enzyme expression, enzyme polymorphisms affecting xenobiotic - mediated toxicity, induction of metabolic enzymes for health benefits, and a diversity of metabolic enzyme modulators.

This detailed book examines the main methods to study mammalian monoamine oxidases (MAOs), ranging from cell biology to computational chemistry. Beginning with techniques on how to obtain pure samples of MAO A and MAO B, the volume continues by covering assays and techniques used to measure MAO enzymatic activity and perform inhibition studies, methods to address cellular localization and function of MAOs, either in cell lines or in animal models, as well as computational methods applied to rational drug design approaches that are used to develop new MAO inhibitors. Written for 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 and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Monoamine Oxidase: Methods and Protocols serves as a vital resource for scientists who are interested in studying MAOs and other similar amine oxidase enzymes.

This book gathers selected studies on the industrial applications of glycoside hydrolases (GHs), presenting an updated classification of these enzymes, and discussing their structure, mechanisms, and various approaches to improve their catalytic efficiency. Further, it explains the various industrial applications of glycoside hydrolases in food, effluent treatment, biofuel production, and the paper and pulp industries. Lastly, the book provides a comparative analysis of glycoside hydrolases and discusses the role of metagenomics in the discovery of industrially important enzymes. As such it is a thought-provoking, instructive and informative resource for biochemists, enzymologists, molecular biologists and bioprocess technologists.

This second edition further develops the principles of applying kinetic principles to drug metabolizing enzymes and transporters. Chapters are divided into six sections detailing fundamental principles of enzyme kinetics, enzyme and transporter structures, highlighting specific oxidative and conjugative drug metabolizing enzymes and drug transporters, modeling approaches for drug metabolizing enzymes and transporters, understanding of variability both experimental and interindividual (pharmacogenomic), and expanded case studies that provide real life examples of applying these principles. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, in some cases step-by-step instructions with readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls with extensive cross referencing to assist in learning. Authoritative and fully updated, Enzyme Kinetics in Drug Metabolism: Fundamentals and Applications, Second Edition serves as a practical teaching tool for novice and advanced scientists interested in the fundamental concepts.

This volume explores a collection of different protocols for the analysis and characterization of DNAzymes and their functions. The topics covered in this book range from bioinformatics and molecular dynamics simulations for the study or modification of nucleic acids to the descriptions of spectroscopic, fluorescence-based, or crystallographic methods to understand the structure and function of DNAzymes. 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 comprehensive, DNAzymes: Methods and Protocols is a valuable resource for scientists and researchers interested in learning more about this evolving field.

This book covers the fundamental aspects of the electrochemistry and redox enzymes that underlie enzymatic bioelectrocatalysis, in which a redox enzyme reaction is coupled with an electrode reaction. Described here are the basic concept and theoretical aspects of bioelectrocatalysis and the various experimental techniques and materials used to study and characterize related problems. Also included are the various applications of bioelectrocatalysis to bioelectrochemical devices including biosensors, biofuel cells, and bioreactors. This book is a unique source of information in the area of enzymatic bioelectrocatalysis, approaching the subject from a cross-disciplinary point of view.

The VIIth International Conference on Aspartic Proteinases was held in Banff, Alberta, Canada, from October 22 to 27, 1996. The venue was the Banff Centre in the Canadian Rockies, a setting well known worldwide for the scenic beauty and mountain grandeur. It was perhaps presumptuous of the organizers to call this the seventh Aspartic Proteinase Conference but it was felt that the meeting in 1982, organized by Tom Blundell and John Kay, was of an international stature and covered topics sufficiently broad to constitute a conference. Thus, there is a discontinuity in that the Gifu Conference organized by Prof. Kenji Takahashi was the fifth International Conference on Aspartic Proteinases. Officially, there has not been a sixth Conference and if there is confusion, it is the result of my desire to recognize the importance of the London meeting. Banffhosted 106 scientists from 14 different countries. There were 26 invited speak ers among the 44 oral presentations of the 7 main sessions. In addition, there were 53 con tributed poster presentations that spanned the whole range of interest in aspartic proteinases.

This volume introduces the reader to the field of enzyme stabilization and the different theories of enzyme stabilization, including the use of immobilization as a stabilization technique. The first part of the book focuses on protocols for enzyme stabilization in solutions including liposome formation, micelle introduction, crosslinking, and additives. The second part of the book discusses protocols for enzyme stabilization during enzyme immobilization, including common techniques like sol-gel encapsulation, polymer encapsulation, and single enzyme nanoparticle formation. Protocols for a variety of enzymes are shown, but the enzymes are chosen as examples to show that these protocols can be used for both enzymes of biological importance, as well as enzymes of industrial importance. The final part details spectroscopic protocols, methods, and assays for studying the effectiveness of the enzyme stabilization and immobilization strategies. 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, Enzyme Stabilization and Immobilization: Methods and Protocols, Second Edition provides molecular biologists, biochemists, and biomedical and biochemical engineers with the state-of-the-art technical information required to effectively stabilize their enzyme of interest in a variety of environments (i.e., harsh temperature, pH, or solvent conditions).

CRISPR-Cas Enzymes, Volume 616, the latest release in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Topics covered in this release include CRISPR bioinformatics, A method for one-step assembly of Class 2 CRISPR arrays, Biochemical reconstitution and structural analysis of ribonucleoprotein complexes in Type I-E CRISPR-Cas systems, Mechanistic dissection of the CRISPR interference pathway in Type I-E CRISPR-Cas system, Site-specific fluorescent labeling of individual proteins within CRISPR complexes, Fluorescence-based methods for measuring target interference by CRISPR-Cas systems, Native State Structural Characterization of CRISRP Associated Complexes using Mass Spectrometry, and more.

Microbes are widely used in large-scale industrial processes due to their versatility, easy growing cultivation, kinetic potential, and the ability to generate metabolites with a wide range of potential applications to various commercial sectors, such as the food, pharmaceutical and cosmetic industries, in addition to the potential for agriculture, biomedical, and several others. Among the metabolites of greatest commercial interest, and many obtained on an industrial scale, the wide range of enzymes, biofuels, organic acids, amino acids, vitamins, biopolymers, and many other classes of metabolites. This book is intended for Bioengineers, Biologist, Biochemist, Biotechnologists, microbiologist, food technologist, enzymologist, and related Professionals/ researchers. Explores recent advances in the valorization of agri-food waste Provides technical concepts on the production of various bio-products of commercial interest Discusses the main process conditions to overcome the difficulties of using waste as alternative raw materials Introduces technical-economic details on the advantages and disadvantages of exploring the waste recovery chain Explores the main technological advances in the recovery of residues in functional products

This definitive reference work describes in detail the enzyme systems that participate in the metabolism of xenobiotics, particularly medicinal drugs. Each chapter focuses on a specific enzyme system, emphasising its role in the activation and detoxication of chemicals. Aspects discussed critically include:

• enzyme function in the metabolism and bioactivation of xenobiotics

• substrate specificity

• tissue distribution

• species distribution (to include laboratory animals and humans)

• hormonal regulation

• sex differences

• modulation by prior exposure to other chemicals

• age-dependent expression

• pharmacogenetics and modulation by disease.

The objective of the Springer Handbook of Enzymes is to provide in concise form data on enzymes sufficiently well characterized. Data sheets are arranged in their EC-Number sequence. Each volume comprises one enzyme class, sometimes the enzyme classes have to be divided into several volumes. Considerable progress has been made in enzymology since the publication of the first edition (published as "Enzyme Handbook): many enzymes are newly classified or reclassified. In the 2nd edition 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). Altogether the amount of data has doubled so that the 2nd edition will consist of approx. 25 volumes. This collection is an indispensable source of information for researchers in biochemistry, biotechnology, organic and analytical chemistry, and food sciences.