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.

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.

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".

This book is a unique resource for state-of-the-art research findings on biotechnological innovations in the area of industrial and therapeutic enzymes, and special-function and extreme-nature enzymes such as ribozymes, therozymes, cold-adapted enzymes, etc, covering all aspects such as the producing micro-organisms, their mode of cultivation, downstream processing and applications. It provides a great deal of information on potential of enzymes for their commercial exploitation. This book gives up-to-date information on advances in enzyme research useful to both the expert and researchers entering the field as well. The vital information has been organized in an easy-to-use format that lets readers become familiar with highlights of the most relevant topics and includes photographs, figures, and tables.

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 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 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.

Authored by the world's leading kinase experts, this is a comprehensive introduction to current knowledge and practice within this emerging field. Following an overview of the major players and pathways that define the kinome, the major part of this work is devoted to current strategies of kinome investigation and manipulation. As such, kinase engineering, peptide substrate engineering, co-substrate design and kinase inhibitor design are discussed in detail, and their potential applications in kinome analysis and kinome-based pharmacotherapy are shown. The result is a toolbox for every kinase researcher: By addressing and comparing current approaches to the study of kinase action, both novice and established researchers will benefit from the practical knowledge contained in this invaluable reference.

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.

This book covers the latest developments in metalloenzymes, including characterizing metal bridging in proteins and peptides, copper(II) complexes of marine peptides, high-spin Co(II) in model and metalloprotein systems to enzymes such as the molybdenum-containing enzymes, CW and pulse EPR of cytochrome P450 enzymes and the radical S-adenosylmethionine FeS family. In the previous two related volumes in the Biological Magnetic Resonance series, High-Resolution EPR: Applications to Metalloenzymes and Metals in Medicine and Metals in Biology:Applications of High-Resolution EPR to Metalloenzymes, topics covered included high-resolution EPR methods, iron proteins, nickel and copper enzymes, metals in medicine, iron-sulfur cluster-containing proteins, and molybdenum enzymes. In this volume, new developments in these areas are covered in detail and new areas that have emerged are also detailed. This is an ideal book for graduate students and researchers working in the fie lds of high-resolution EPR, metalloenzymes, and metals in biology.

This book provides an account of the recent advances in our understanding of the role of proteases under physiological and pathological conditions. It reviews the contributions that have been made in the fields of biochemistry, molecular biology, medicine, agriculture and ecology. In addition there is an account of the growing number of practical applications in biotechnology, reflecting the fact that peptidases are major targets for medical and agricultural purposes. The book shows how the numerous protease structures are essential for drug design. The contributions place special emphasis on cysteine, aspartic and metalloproteases and their role in physiological and certain pathological states. Another focus is the classification and nomenclature of peptidases and a review of those proteases currently under the most intensive investigation. The book provides an informative introduction for teachers and newcomers to the field, such as graduate students, while providing a valuable source of material and ideas for the academic and industrial researcher in areas including biochemistry, medicine, agriculture and biotechnology.

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).

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.

This edited work presents studies that clarify the basics of producing recombinant enzymes that finally lead to commercialization. It enables researchers to see what is crucial to the commercialization process, from examining the cloning method, using analytical techniques such as calculating the total protein content and enzyme activity, through considering upstream and downstream processes, to the final product. Readers will discover the importance of the cloning method as it influences the upstream and downstream processes and determines the level of success of the recombinant enzyme commercialization processes. We see that the two main factors that are particularly sensitive during the cloning process are the vector and the host. A discussion of analytical techniques is presented followed by studies on important stages during the upstream processes including the process of optimizing the media to get results and high enzyme activity. Downstream processes such as the cell disruption technique, purification and formulation of the final product are then considered. The reader is introduced to software that helps streamline recombinant enzyme production from the upstream to downstream processes, to facilitate the process of up-scaling production. This work includes a case study as tool, to guide understanding of the commercialization process. The work is written for researchers in the field and is especially suited to those who are under pressure to embark on the tough process of commercialization.

This volume summarizes our current knowledge on different biomass-converting enzymes and their potential use in converting biomass into simple sugar to generate bioenergy and other value added co-/by products. It consists of 13 chapters and is divided into three parts: cellulases; hemicellulases; and lignocellulose oxidoreductases. Written by international experts, the contributions offer clear and concise descriptions of both standard and new technologies. It is an invaluable reference resource for undergraduates, post-graduates, researchers and practitioners in the field of microbial enzymes for biofuel and biorefinery applications.

Metalloproteins are involved in many key biological processes such as gas transport and metabolism, photosynthesis, cell respiration, the Krebs cycle and many other vital redox reactions. Metalloproteins: Methods and Protocols addresses multiple aspects of metalloenzyme research from the production and purification of metalloproteins using both standard recombinant techniques and a cell-free system, to the electrochemical analysis of metalloproteins, as well as IR techniques, Moessbauer spectroscopy and the theoretical interpretation of metalloenzyme catalytic and redox processes, to name a few topics. Written in the 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 protocols and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Metalloproteins: Methods and Protocols seeks to serve both professionals and novices interested in the metalloprotein field.

This book provides a review of the multitude of nucleic acid polymerases, including DNA and RNA polymerases from Archea, Bacteria and Eukaryota, mitochondrial and viral polymerases, and other specialized polymerases such as telomerase, template-independent terminal nucleotidyl transferase and RNA self-replication ribozyme. Although many books cover several different types of polymerases, no book so far has attempted to catalog all nucleic acid polymerases. The goal of this book is to be the top reference work for postgraduate students, postdocs, and principle investigators who study polymerases of all varieties. In other words, this book is for polymerase fans by polymerase fans.

Nucleic acid polymerases play a fundamental role in genome replication, maintenance, gene expression and regulation. Throughout evolution these enzymes have been pivotal in transforming life towards RNA self-replicating systems as well as into more stable DNA genomes. These enzymes are generally extremely efficient and accurate in RNA transcription and DNA replication and share common kinetic and structural features. How catalysis can be so amazingly fast without loss of specificity is a question that has intrigued researchers for over 60 years. Certain specialized polymerases that play a critical role in cellular metabolism are used for diverse biotechnological applications and are therefore an essential tool for research.

This book provides a selection of recently developed methods and protocols in bacterial glycomics to aid in bettering our understanding of the structures and functions of bacterial polysaccharides, their attachments to proteins and lipids, their role in biofilm formation, as well as their biosynthesis. With the emerging bacterial resistance to commonly used antibiotics world-wide, these techniques to study the outer polysaccharides of bacteria, with their functions in bacterial adhesion, colonization, growth, establishment of biofilms, and control virulence and pathogenicity, are increasingly important. 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 cutting-edge, Bacterial Polysaccharides: Methods and Protocols aims to support researchers contributing to future approaches that will fill our knowledge gaps and define anti-bacterial targets.

Enzyme Technology is one the most promising disciplines in modern biotechnology. In this book, the applications of a wide variety of enzymes are highlighted. Current studies in enzyme technology are focused towards the discovery of novel enzymes (termed "bio-discovery" or "bio-prospecting") and the identification and elucidation of novel pathways of these novel enzymes with emphasis on their industrial relevance. With the development of molecular techniques and other bioinformatics tools, the time to integrate this subject with other fields in the life sciences has arrived. A rapid expansion of the knowledge base in the field of enzyme biotechnology has occurred over the past few years. Much of this expansion has been driven by the bio-discovery of many new enzymes from a wide range of environments, some extreme in nature, followed by subsequent protein (enzyme) engineering. These enzymes have found a wide range of applications, ranging from bioremediation, bio-monitoring, biosensor development, bioconversion to biofuels and other biotechnologically important value-added products. Hydrolases constitute a major component of the global annual revenue generated by industrial enzymes and the emphasis has therefore been placed on these enzymes and their applications. With the immense interest of researchers active in this area, this book will serve to provide information on current aspects in this field of study. In the current edition, the contributions of many diversified topics towards establishing new directions of research in the area of enzyme biotechnology are described. This book serves to provide a unique source of information to undergraduates, post graduates and doctoral courses in microbiology and biotechnology along with allied life sciences. The present edition of the book covers all important areas of enzyme biotechnology i.e. the wide variety of enzymes in the field of enzyme biotechnology and their industrial applications, new methods and state-of-the-art information on modern methods of enzyme discovery. This book will act as good resource on most of the current facets of enzyme technology for all students engaged in bioengineering and biotechnology.

This thesis describes a series of investigations designed to assess the value of metalloenzymes in systems for artificial and adapted photosynthesis. The research presented explores the interplay between inherent enzyme properties such as structure, rates and thermodynamics, and the properties of the semiconducting materials to which the enzyme is attached. Author, Andreas Bachmeier provides a comprehensive introduction to the interdisciplinary field of artificial photosynthesis, allowing the reader to grasp the latest approaches being investigated, from molecular systems to heterogeneous surface catalysis. Bachmeier's work also uses metalloenzymes to highlight the importance of reversible catalysts in removing the burden of poor electrocatalytic rates and efficiencies which are common characteristics for most artificial photosynthesis systems. Overall, this thesis provides newcomers and students in the field with evidence that metalloenzymes can be used to establish new directions in artificial photosynthesis research.

This volume presents an overview of nonribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) structure and function. It then continues with methods for the analysis of these pathways including conventional enzymological assays, contemporary mass spectrometric analysis techniques, specialized molecular biological approaches applicable to NRPSs and PKSs, and small molecule analysis tools tailored to this very special class of natural products, and concludes by examining bioinformatics tools for the analysis of these enzymes, pathways, and molecules. 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, Nonribosomal Peptide and Polyketide Biosynthesis: Methods and Protocols serves as a valuable reference for those experienced in studying NRPS and PKS enzymes, pathways, and natural products as well as a gateway for those just entering the field.