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| enzyme engineering methods and protocols: Enzyme Engineering Francesca Magnani, Chiara Marabelli, Francesca Paradisi, 2021-11-24 This book provides comprehensive methods and protocols about enzyme design. The chapters are grouped by main topic, starting with methodologies describing library preparation and screening, state of the art techniques in directed evolution and rational design, followed by examples of immobilization of enzymes on sustainable polymers, as well as biocatalytic conversions mediated by homogenous enzymatic preparations or whole cells. 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 timely, Enzyme Engineering: Methods and Protocols is an ideal guide for both the novice and the veteran researcher interested in biocatalysis. Chapter 13 is available open access under a CC BY 4.0 license. |
| enzyme engineering methods and protocols: Enzyme Engineering James Samuelson, 2016-05-01 This book provides guidance to those wishing to create enzyme variants. It covers such topics as a simple method for generating site-specific mutations within bacterial chromosomes and the engineering of two difference types of rare-cutting endonucleases. |
| enzyme engineering methods and protocols: Enzyme Engineering Francesca Magnani, Chiara Marabelli, Francesca Paradisi, 2022 This book provides comprehensive methods and protocols about enzyme design. The chapters are grouped by main topic, starting with methodologies describing library preparation and screening, state of the art techniques in directed evolution and rational design, followed by examples of immobilization of enzymes on sustainable polymers, as well as biocatalytic conversions mediated by homogenous enzymatic preparations or whole cells. 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 timely, Enzyme Engineering: Methods and Protocols is an ideal guide for both the novice and the veteran researcher interested in biocatalysis. Chapter 13 is available open access under a CC BY 4.0 license. |
| enzyme engineering methods and protocols: Microbial Metabolic Engineering Christine Nicole S. Santos, Parayil Kumaran Ajikumar, 2019-02-21 This volume covers a wide array of topics that will aid researchers in the task of engineering complex biological systems. This book is divided into three parts: Part One discusses the discovery and identification of relevant biosynthetic pathways for engineering; Part Two looks at the development of genetic tools for manipulating enzymes, biosynthetic pathways, and whole genomes; and Part Three covers the characterization of engineered microbes using targeted and global systems biology tools, as well as in silico models. Chapters explore topics such as leveraging enzyme promiscuity to construct novel biosynthetic pathways; assembling combinatorial multigene pathways for rapid strain optimization; applying 'omics technologies for identifying bottlenecks; and engineering nontraditional host organisms like cyanobacterium and Yarrowia lipolytica. 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 authoritative, Microbial Metabolic Engineering: Methods and Protocols is a valuable resource for researchers and scientists interested in engineering and optimizing microbes for a variety of biotechnological applications. |
| enzyme engineering methods and protocols: Protein Engineering Uwe T. Bornscheuer, Matthias Höhne, 2017-10-30 This volume details basic and advanced protocols for both stages of protein engineering: the library design phase and the identification of improved variants by screening and selection. Chapters focus on enzyme engineering using rational and semi-rational approaches. 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. Authoritative and cutting-edge, Protein Engineering: Methods and Protocols aims to aid scientists in the planning and performance of their experiments. The chapter 'Functional Analysis of Membrane Proteins Produced by Cell-Free Translation' is open access under a CC BY 4.0 license via link.springer.com. |
| enzyme engineering methods and protocols: Directed Enzyme Evolution Frances H. Arnold, George Georgiou, 2008-02-02 Directed evolution comprises two distinct steps that are typically applied in an iterative fashion: (1) generating molecular diversity and (2) finding among the ensemble of mutant sequences those proteins that perform the desired fu- tion according to the specified criteria. In many ways, the second step is the most challenging. No matter how cleverly designed or diverse the starting library, without an effective screening strategy the ability to isolate useful clones is severely diminished. The best screens are (1) high throughput, to increase the likelihood that useful clones will be found; (2) sufficiently sen- tive (i. e. , good signal to noise) to allow the isolation of lower activity clones early in evolution; (3) sufficiently reproducible to allow one to find small improvements; (4) robust, which means that the signal afforded by active clones is not dependent on difficult-to-control environmental variables; and, most importantly, (5) sensitive to the desired function. Regarding this last point, almost anyone who has attempted a directed evolution experiment has learned firsthand the truth of the dictum “you get what you screen for. ” The protocols in Directed Enzyme Evolution describe a series of detailed p- cedures of proven utility for directed evolution purposes. The volume begins with several selection strategies for enzyme evolution and continues with assay methods that can be used to screen enzyme libraries. Genetic selections offer the advantage that functional proteins can be isolated from very large libraries s- ply by growing a population of cells under selective conditions. |
| enzyme engineering methods and protocols: Yeast Metabolic Engineering Valeria Mapelli, 2016-08-23 Yeast Metabolic Engineering: Methods and Protocols provides the widely established basic tools used in yeast metabolic engineering, while describing in deeper detail novel and innovative methods that have valuable potential to improve metabolic engineering strategies in industrial biotechnology applications. Beginning with an extensive section on molecular tools and technology for yeast engineering, this detailed volume is not limited to methods for Saccharomyces cerevisiae, but describes tools and protocols for engineering other yeasts of biotechnological interest, such as Pichia pastoris, Hansenula polymorpha and Zygosaccharomyces bailii. Tools and technologies for the investigation and determination of yeast metabolic features are described in detail as well as metabolic models and their application for yeast metabolic engineering, while a chapter describing patenting and regulations with a special glance at yeast biotechnology closes the volume. Written in the highly successful Methods in Molecular Biology series format, most chapters include an introduction to their respective topic, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and tips on troubleshooting and avoiding known pitfalls. Comprehensive and authoritative, Yeast Metabolic Engineering: Methods and Protocols aims to familiarize researchers with the current state of these vital and increasingly useful technologies. |
| enzyme engineering methods and protocols: Fundamentals of Enzyme Engineering Young Je Yoo, Yan Feng, Yong-Hwan Kim, Camila Flor J. Yagonia, 2017-01-12 This book provides a comprehensive introduction to all aspects of enzyme engineering, from fundamental principles through to the state-of-the-art in research and industrial applications. It begins with a brief history, describing the milestones of advancement in enzyme science and technology, before going on to cover the fundamentals of enzyme chemistry, the biosynthesis of enzymes and their production. Enzyme stability and the reaction kinetics during enzymatic reactions are presented to show how enzymes function during catalysis and the factors that affect their activity. Methods to improve enzyme performance are also presented, such as cofactor regeneration and enzyme immobilization. The book emphasizes and elaborates on the performance and characteristics of enzymes at the molecular level. Finally, the book presents recent advances in enzyme engineering and some key industrial application of enzymes addressing the present needs of society. This book presents essential information not only for undergraduate and graduate students, but also for researchers in academia and industry, providing a valuable reference for the development of commercial applications of enzyme technology. |
| enzyme engineering methods and protocols: Protein Engineering Matthias Höhne, Uwe T. Bornscheuer, 2018 This volume details basic and advanced protocols for both stages of protein engineering: the library design phase and the identification of improved variants by screening and selection. Chapters focus on enzyme engineering using rational and semi-rational approaches. 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. Authoritative and cutting-edge, Protein Engineering: Methods and Protocols aims to aid scientists in the planning and performance of their experiments. |
| enzyme engineering methods and protocols: Enzyme Stabilization and Immobilization Shelley D. Minteer, 2016-10-22 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). |
| enzyme engineering methods and protocols: Immobilization of Enzymes and Cells Jose M. Guisan, Juan M. Bolivar, Fernando López-Gallego, Javier Rocha-Martín, 2021-01-29 This fourth edition volume expands on the previous editions with new insights on important aspects to take into accounting when immobilizing enzymes and cells, illustrating outstanding examples that support those aspects, and exploring ways to fabricate and characterize heterogeneous biocatalysts including both immobilized enzymes and cells. The transformation of soluble and usually instable enzymes into heterogeneous and highly stable biocatalysts is strongly emphasized. The chapters in this book cover topics such as the importance of enzyme orientation on the support surface; application and characterization of immobilized enzymes; different functionalization chemistries for the modulation of the immobilized enzyme properties; co-immobilization of multi-enzyme systems; new analytical techniques for the characterization of heterogeneous biocatalysts; protocols for cell entrapment in alginate; preparation and characterization of biofilms; and cell encapsulation technologies. 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 authoritative, Immobilization of Enzymes and Cells: Methods and Protocols, Fourth Edition is a valuable resource for researchers interested in expanding their knowledge of this developing field. |
| enzyme engineering methods and protocols: Protocols and Applications in Enzymology Seema Anil Belorkar, Sudisha Jogaiah, 2021-11-24 Protocols and Applications in Enzymology provides instruction on the experimental procedures of enzyme isolation techniques, innovative screening techniques, and instrument enabled enzyme assays and their underlying principles, among other protocols. The book serves as a one-stop solution for those working with different enzyme protocols in the fields of biochemistry, microbiology, biotechnology and allied subjects. Each chapter offers a full overview of protocol key resources, materials required, quantifiable and statistical analysis, optimization and troubleshooting, safety considerations, and standards. Applications are discussed across distribution and diversity of microbial enzymes, enzyme screening, enzymes in solid state fermentations, enzyme assays, enzyme kinetics, and biotechnological uses. Provides step-by-step instruction on enzyme protocols and applications, with actionable discussions of needed resources, materials, quantification and statistical analysis, optimization and troubleshooting, safety considerations and standards Presents easy to read, reproducible protocols for researchers and students across academia and industry Includes color diagrams that illustrate key concepts |
| enzyme engineering methods and protocols: Biocatalysis for Practitioners Gonzalo de Gonzalo, Iván Lavandera, 2021-07-19 This reference book originates from the interdisciplinary research cooperation between academia and industry. In three distinct parts, latest results from basic research on stable enzymes are explained and brought into context with possible industrial applications. Downstream processing technology as well as biocatalytic and biotechnological production processes from global players display the enormous potential of biocatalysts. Application of extreme reaction conditions (i.e. unconventional, such as high temperature, pressure, and pH value) - biocatalysts are normally used within a well defined process window - leads to novel synthetic effects. Both novel enzyme systems and the synthetic routes in which they can be applied are made accessible to the reader. In addition, the complementary innovative process technology under unconventional conditions is highlighted by latest examples from biotech industry. |
| enzyme engineering methods and protocols: Enzyme Engineering and Evolution: General Methods , 2020-09-05 This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. - Provides the authority and expertise of leading contributors from an international board of authors - Presents the latest release in the Methods in Enzymology series |
| enzyme engineering methods and protocols: Noncanonical Amino Acids Edward A. Lemke, 2018 |
| enzyme engineering methods and protocols: Directed Enzyme Evolution: Advances and Applications Miguel Alcalde, 2017-02-14 This book focuses on some of the most significant advances in enzyme engineering that have been achieved through directed evolution and hybrid approaches. On the 25th anniversary of the discovery of directed evolution, this volume is a tribute to the pioneers of this thrilling research field, and at the same time provides a comprehensive overview of current research and the state of the art. Directed molecular evolution has become the most reliable and robust method to tailor enzymes, metabolic pathways or even whole microorganisms with improved traits. By mirroring the Darwinian algorithm of natural selection on a laboratory scale, new biomolecules of invaluable biotechnological interest can now be engineered in a manner that surpasses the boundaries of nature. The volume is divided into two sections, the first of which provides an update on recent successful cases of enzyme ensembles from different areas of the biotechnological spectrum, including tryptophan synthases, unspecific peroxygenases, phytases, therapeutic enzymes, stereoselective enzymes and CO2-fixing enzymes. This section also provides information on the directed evolution of whole cells. The second section of the book summarizes a variety of the most applicable methods for library creation, together with the future trends aimed at bringing together directed evolution and in silico/computational enzyme design and ancestral resurrection. |
| enzyme engineering methods and protocols: Synthetic Metabolic Pathways Michael Krogh Jensen, Jay D. Keasling, 2017-11-24 This volume outlines key steps associated with the design, building, and testing of synthetic metabolic pathways for optimal cell factory performance and robustness, and illustrates how data-driven learning from these steps can be used for rational cost-effective engineering of cell factories with improved performance. Chapters are divided into four sections focusing on the four steps of the iterative design-build-test-learn cycle related to modern cell factory engineering. 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. Authoritative and practical, Synthetic Metabolic Pathways: Methods and Protocols aims to ensure successful results in the further study of this vital field. |
| enzyme engineering methods and protocols: Protein Engineering Methods and Protocols Peter W. Goodenough, 1998-10 |
| enzyme engineering methods and protocols: Protein Affinity Tags Richard J. Giannone, Andrew B. Dykstra, 2014-06-19 Protein Affinity Tags: Methods and Protocols provides researchers with the necessary information, tools, and strategy required for proper inquiry into a given protein’s function and structure. Today’s researchers can easily alter the amino acid sequence of any given protein, a powerful technology allowing for the precise engineering of specific proteins. Protein affinity tagging employs the use of known protein binding interactions in order to “fish out” a protein-of-interest or, with more advanced tags, a protein complex. This technology paired with recent advancements in DNA sequencing technology promises a future that is awash with novel genomic information – information that will not only expand our knowledge of the processes that govern life, but also fuel innovation that will undoubtedly conquer many of today’s most salient health and environmental challenges. 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, Protein Affinity Tags: Methods and Protocols will be an invaluable resource to those seeking to employ protein affinity tags to study their biological system of interest. |
| enzyme engineering methods and protocols: Enzyme Engineering Manfred T. Reetz, Zhoutong Sun, Ge Qu, 2023-01-10 Enzyme Engineering An authoritative and up-to-date discussion of enzyme engineering and its applications In Enzyme Engineering: Selective Catalysts for Applications in Biotechnology, Organic Chemistry, and Life Science, a team of distinguished researchers deliver a robust treatment of enzyme engineering and its applications in various fields such as biotechnology, life science, and synthesis. The book begins with an introduction to different protein engineering techniques, covers topics like gene mutagenesis methods for directed evolution and rational enzyme design. It includes industrial case studies of enzyme engineering with a focus on selectivity and activity. The authors also discuss new and innovative areas in the field, involving machine learning and artificial intelligence. It offers several insightful perspectives on the future of this work. Readers will also find: A thorough introduction to directed evolution and rational design as protein engineering techniques Comprehensive explorations of screening and selection techniques, gene mutagenesis methods in directed evolution, and guidelines for applying gene mutagenesis in organic chemistry, pharmaceutical applications, and biotechnology Practical discussions of protein engineering of enzyme robustness relevant to organic and pharmaceutical chemistry Treatments of artificial enzymes as promiscuous catalysts Various lessons learned from semi-rational and rational directed evolution A transdisciplinary treatise, Enzyme Engineering: Selective Catalysts for Applications in Biotechnology, Organic Chemistry, and Life Science is perfect for protein engineers, theoreticians, organic, and pharmaceutical chemists as well as transition metal researchers in catalysis and biotechnologists. |
| enzyme engineering methods and protocols: Enzymes in Nonaqueous Solvents Evgeny N. Vulfson, 2010-11-19 Enzymatic catalysis has gained considerable attention in recent years as an efficient tool in the preparation of natural products, pharmaceuticals, fine chemicals, and food ingredients. The high selectivity and mild reaction con- tions associated with enzymatic transformations have made this approach an attractive alternative in the synthesis of complex bioactive compounds, which are often difficult to obtain by standard chemical routes. However, the maj- ity of organic compounds are not very soluble in water, which was traditi- ally perceived as the only suitable reaction medium for the application of biocatalysts. The realization that most enzymes can function perfectly well under nearly anhydrous conditions and, in addition, display a number of useful properties, e. g. , highly enhanced stability and different selectivity, has d- matically widened the scope of their application to the organic synthesis. Another great attraction of using organic solvents rather than water as a reaction solvent is the ability to perform synthetic transformations with re- tively inexpensive hydrolytic enzymes. It is worth reminding the reader that in vivo, the synthetic and hydrolytic pathways are catalyzed by different enzymes. However, elimination of water from the reaction mixture enables the “reversal” of hydrolytic enzymes and thus avoids the use of the expensive cofactors or activated substrates that are required for their synthetic count- parts. |
| enzyme engineering methods and protocols: Unnatural Amino Acids Loredano Pollegioni, Stefano Servi, 2011-10-05 Even though they are present in nature, non-proteinogenic amino acids are usually defined as unnatural or non-natural. Beside their structural diversity, interest in these compounds is due to their occurrence in nature, their biological properties, the analytical aspects, their use as probes, and their incorporation into peptides and proteins, among other reasons. Divided into five convenient sections, Unnatural Amino Acids: Methods and Protocols deals with enzymatic methods used to produce non-natural amino acids, aspects concerning the presence of unnatural amino acids in peptides with antimicrobial properties, genetic incorporation of unnatural amino acids into proteins (yeast and mammalian cells), and detection and quantification of D-amino acids and related enzymes. Written in the highly successful Methods in Molecular BiologyTM 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. Authoritative and accessible, Unnatural Amino Acids: Methods and Protocols serves as an ideal guide for scientists and contributes to directing the attention of researchers to the many fields of growing scientific interest in non-natural amino acids. |
| enzyme engineering methods and protocols: Modern Biocatalysis Gavin Williams, Mélanie Hall, 2018-05-31 The synergy between synthetic biology and biocatalysis is emerging as an important trend for future sustainable processes. This book reviews all modern and novel techniques successfully implemented in biocatalysis, in an effort to provide better performing enzymatic systems and novel biosynthetic routes to (non-)natural products. This includes the use of molecular techniques in protein design and engineering, construction of artificial metabolic pathways, and application of computational methods for enzyme discovery and design. Stress is placed on current 'hot' topics in biocatalysis, where recent advances in research are defining new grounds in enzyme-catalyzed processes. With contributions from leading academics around the world, this book makes a ground-breaking contribution to this progressive field and is essential reading for graduates and researchers investigating (bio)catalysis, enzyme engineering, chemical biology, and synthetic biology. |
| enzyme engineering methods and protocols: In Vitro Mutagenesis Protocols Jeff Braman, 2008-02-05 Hands-on researchers with proven track records describe in stepwise fashion their advanced mutagenesis techniques. The contributors focus on improvements to conventional site-directed mutagenesis, including a chapter on chemical site-directed mutagenesis, PCR-based mutagenesis and the modifications that allow high throughput mutagenesis experiments, and mutagenesis based on gene disruption (both in vitro- and in situ-based). Additional methods are provided for in vitro gene evolution; for gene disruption based on recombination, transposon, and casette mutagenesis; and for facilitating the introduction of multiple mutations. Time-tested and highly practical, the protocols in In Vitro Mutagenesis Protocols, 2nd Edition offer today's molecular biologists reliable and powerful techniques with which to illuminate the proteome. |
| enzyme engineering methods and protocols: Plant Metabolism Ganesh Sriram, 2013-11-12 In Plant Metabolism: Methods and Protocols, expert researchers in the field present the latest methods on quantitative analysis of plant metabolism. The methods focus on measurements, analyses and simulations of molecules, fluxes, and ultimately entire metabolic pathways and networks. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials, reagents, or software, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Plant Metabolism: Methods and Protocols seeks to benefit scientists ranging from plant biology, metabolic engineering, and biotechnology. |
| enzyme engineering methods and protocols: Protein Engineering Huimin Zhao, 2021-08-23 A one-stop reference that reviews protein design strategies to applications in industrial and medical biotechnology Protein Engineering: Tools and Applications is a comprehensive resource that offers a systematic and comprehensive review of the most recent advances in the field, and contains detailed information on the methodologies and strategies behind these approaches. The authors—noted experts on the topic—explore the distinctive advantages and disadvantages of the presented methodologies and strategies in a targeted and focused manner that allows for the adaptation and implementation of the strategies for new applications. The book contains information on the directed evolution, rational design, and semi-rational design of proteins and offers a review of the most recent applications in industrial and medical biotechnology. This important book: Covers technologies and methodologies used in protein engineering Includes the strategies behind the approaches, designed to help with the adaptation and implementation of these strategies for new applications Offers a comprehensive and thorough treatment of protein engineering from primary strategies to applications in industrial and medical biotechnology Presents cutting edge advances in the continuously evolving field of protein engineering Written for students and professionals of bioengineering, biotechnology, biochemistry, Protein Engineering: Tools and Applications offers an essential resource to the design strategies in protein engineering and reviews recent applications. |
| enzyme engineering methods and protocols: Pichia Protocols James M Cregg, 2007-08-08 This book focuses on recent developments of Pichia pastoris as a recombinant protein production system. Highlighted topics include a discussion on the use of fermentors to grow Pichia pastoris, information on the O- and N-linked glycosylation, methods for labeling Pichia pastoris expressed proteins for structural studies, and the introduction of mutations in Pichia pastoris genes by the methods of restriction enzyme-mediated integration (REMI). Each chapter presents cutting-edge and cornerstone protocols for utilizing P. pastoris as a model recomibinant protein production system. This volume fully updates and expands upon the first edition. |
| enzyme engineering methods and protocols: Downstream Processing of Proteins Mohamed A. Desai, 2008-02-05 Considerable effort and time is allocated to introducing cell culture and fermentation technology to undergraduate students in academia, generally through a range of courses in industrial biotechnology and related disciplines. Similarly, a large number of textbooks are available to describe the appli- tions of these technologies in industry. However, there has been a general lack of appreciation of the significant developments in downstream processing and isolation technology, the need for which is largely driven by the stringent re- latory requirements for purity and quality of injectable biopharmaceuticals. This is particularly reflected by the general absence of coverage of this s- ject in many biotechnology and related courses in educational institutions. For a considerable while I have felt that there is increasing need for an introductory text to various aspects of downstream processing, particularly with respect to the needs of the biopharmaceutical and biotechnology ind- try. Although there are numerous texts that cover various aspects of protein purification techniques in isolation, there is a need for a work that covers the broad range of isolation technology in an industrial setting. It is anticipated that Downstream Processing of Proteins: Methods and Protocols will play a small part in filling this gap and thus prove a useful contribution to the field. It is also designed to encourage educational strategists to broaden the coverage of these topics in industrial biotechnology courses by including accounts of this important and rapidly developing element of the industrial process. |
| enzyme engineering methods and protocols: Techniques in Genetic Engineering Isil Aksan Kurnaz, 2015-05-08 Although designed for undergraduates with an interest in molecular biology, biotechnology, and bioengineering, this book-Techniques in Genetic Engineering-IS NOT: a laboratory manual; nor is it a textbook on molecular biology or biochemistry. There is some basic information in the appendices about core concepts such as DNA, RNA, protein, genes, and |
| enzyme engineering methods and protocols: Protocols and Applications in Enzymology Seema Anil Belorkar, Sudisha Jogaiah, 2021-11-19 Protocols and Applications in Enzymology provides instruction on the experimental procedures of enzyme isolation techniques, innovative screening techniques, and instrument enabled enzyme assays and their underlying principles, among other protocols. The book serves as a one-stop solution for those working with different enzyme protocols in the fields of biochemistry, microbiology, biotechnology and allied subjects. Each chapter offers a full overview of protocol key resources, materials required, quantifiable and statistical analysis, optimization and troubleshooting, safety considerations, and standards. Applications are discussed across distribution and diversity of microbial enzymes, enzyme screening, enzymes in solid state fermentations, enzyme assays, enzyme kinetics, and biotechnological uses. - Provides step-by-step instruction on enzyme protocols and applications, with actionable discussions of needed resources, materials, quantification and statistical analysis, optimization and troubleshooting, safety considerations and standards - Presents easy to read, reproducible protocols for researchers and students across academia and industry - Includes color diagrams that illustrate key concepts |
| enzyme engineering methods and protocols: Directory of Therapeutic Enzymes Barry M. McGrath, Gary Walsh, 2005-08-12 Carefully crafted to provide tightly focused and authoritative information, the Directory of Therapeutic Enzymes covers all approved therapeutic enzymes currently used in medicine. Written mainly by industry experts, the book includes information sourced directly from the company that developed or manufactured the product. It explores major |
| enzyme engineering methods and protocols: Yeast Surface Display Bin Liu, 2016-10-09 In addition to research and discovery, yeast surface display technology has found applications in industrial processes such as biofuel production and environmental pollutant absorption and degradation. Yeast Surface Display: Methods, Protocols, and Applications guides readers through yeast surface antibody display library and antibody engineering, yeast surface display as a tool for protein engineering, yeast surface cDNA display library construction and applications, and yeast surface display in bioassay and industrial applications. 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 key tips on troubleshooting and avoiding known pitfalls. Concise and easy-to-use, Yeast Surface Display: Methods, Protocols, and Applications aims to help accelerate the work of protein chemists, antibody engineers, molecular and cell biologists, and industrial bioengineers. |
| enzyme engineering methods and protocols: Applied Bioengineering Toshiomi Yoshida, 2017-06-06 A comprehensive overview of the topic, highlighting recent developments, ongoing research trends and future directions. Experts from Europe, Asia and the US cover five core areas of imminent importance to the food, feed, pharmaceutical and water treatment industries in terms of sustainable and innovative processing and production. In the field of enzyme engineering, they summarize historic developments and provide an overview of molecular enzyme engineering, while also discussing key principles of microbial process engineering, including chapters on process development and control. Further sections deal with animal and plant cell culture engineering. The final section of the book deals with environmental topics and highlights the application of bioengineering principles in waste treatment and the recovery of valuable resources. With its cutting-edge visions, extensive discussions and unique perspectives, this is a ready reference for biotechnologists, bioengineers, bioengineers, biotechnological institutes, and environmental chemists. |
| enzyme engineering methods and protocols: Cell-Free Protein Production Yaeta Endo, Kazuyuki Takai, Takuya Ueda, 2009-12-04 During the past decade as the data on gene sequences and expression patterns rapidly accumulated, cell-free protein synthesis technology has also experienced a revolution, becoming a powerful tool for the preparation of proteins for their functional and structural analysis. In Cell-Free Protein Production: Methods and Protocols, experts in the field contribute detailed techniques, the uses of which expand deep into the studies of biochemistry, molecular biology, and biotechnology. Beginning briefly with basic methods and historical aspects, the book continues with thorough coverage of protein preparation methods, the preparation of proteins that are generally difficult to prepare in their functional forms, applications of the cell-free technologies to protein engineering, as well as some methods that are expected to constitute a part of future technologies. Written in the highly successful Methods in Molecular BiologyTM series format, the chapters include 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. Authoritative and cutting-edge, Cell-Free Protein Production: Methods and Protocols aims to help researchers continue the growth of the vital exploration of cell-free sciences and technologies in order to better understand the dynamic lives of cells. |
| enzyme engineering methods and protocols: Enzyme Engineering and Evolution: Specific Enzyme Applications , 2020-09-15 Methods in Enzymology, Volume 644, the latest release in this ongoing serial, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Chapters in this new release include Site-directed recombination (SDR) in vivo: a fast and reliable tool to unveil beneficial epistasis, Creation and application of amine oxidase with expanded substrate specificities from porcine kidney D-amino acid oxidase, Methods to assess correlation networks for engineering transketolase, Exploration of Enzyme Diversity by Integrating Bioinformatics with Microfluidics, Engineering lytic polysaccharide monooxygenases (LPMOs), Emulsion-based directed evolution of enzymes in yeast, and much more. - Provides the authority and expertise of leading contributors from an international board of authors - Presents the latest release in the Methods in Enzymology series |
| enzyme engineering methods and protocols: The Cannabis Genome Angel Fernandez i Marti, Jaime Prohens, 2025-06-17 Cannabis is undergoing a remarkable transformation—from a misunderstood plant to a cornerstone of modern medicine, industry, and agriculture. This book bridges the gap between tradition and innovation, providing a holistic exploration of cannabis genetics, breeding, genomics, and its wide-ranging applications. Authored by leading researchers, industry pioneers, and legacy breeders, it offers a unique blend of academic rigor and practical insights. Unlike other resources, this book integrates perspectives from public and private sectors, honoring those who have advanced cannabis cultivation despite legal and social barriers. It recognizes the pivotal role of startups like MyFloraDNA, which invest heavily in groundbreaking research, shaping the future of this extraordinary plant. Key Features: Comprehensive insights into cannabis origins, taxonomy, and genetic diversity. Advanced breeding techniques and applications of tissue culture, CRISPR, and omics technologies. Practical strategies for pest management, crop improvement, and intellectual property protection. Forward-looking perspectives on cannabis in personalized medicine, sustainability, and global agriculture. Written for scientists, growers, policymakers, and enthusiasts, this book addresses the economic, social, and scientific dimensions of cannabis. It offers actionable knowledge to drive progress in cultivation, industry, and research. |
| enzyme engineering methods and protocols: Trichoderma Reesei Astrid R. Mach-Aigner, Roland Martzy, 2021 |
| enzyme engineering methods and protocols: Protein Engineering Protocols Kristian Müller, Katja Arndt, 2007-10-26 Protein engineering is a fascinating mixture of molecular biology, protein structure analysis, computation, and biochemistry, with the goal of developing useful or valuable proteins. Protein Engineering Protocols will consider the two general, but not mutually exclusive, strategies for protein engineering. The first is known as rational design, in which the scientist uses detailed knowledge of the structure and function of the protein to make desired changes. The s- ond strategy is known as directed evolution. In this case, random mutagenesis is applied to a protein, and selection or screening is used to pick out variants that have the desired qualities. By several rounds of mutation and selection, this method mimics natural evolution. An additional technique known as DNA shuffling mixes and matches pieces of successful variants to produce better results. This process mimics recombination that occurs naturally during sexual reproduction. The first section of Protein Engineering Protocols describes rational p- tein design strategies, including computational methods, the use of non-natural amino acids to expand the biological alphabet, as well as impressive examples for the generation of proteins with novel characteristics. Although procedures for the introduction of mutations have become routine, predicting and und- standing the effects of these mutations can be very challenging and requires profound knowledge of the system as well as protein structures in general. |
| enzyme engineering methods and protocols: Non-Natural Amino Acids , 2009-07-24 By combining the tools of organic chemistry with those of physical biochemistry and cell biology, Non-Natural Amino Acids aims to provide fundamental insights into how proteins work within the context of complex biological systems of biomedical interest. The critically acclaimed laboratory standard for 40 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. With more than 400 volumes published, each Methods in Enzymology volume presents material that is relevant in today's labs -- truly an essential publication for researchers in all fields of life sciences. - Demonstrates how the tools and principles of chemistry combined with the molecules and processes of living cells can be combined to create molecules with new properties and functions found neither in nature nor in the test tube - Presents new insights into the molecular mechanisms of complex biological and chemical systems that can be gained by studying the structure and function of non-natural molecules - Provides a one-stop shop for tried and tested essential techniques, eliminating the need to wade through untested or unreliable methods |
| enzyme engineering methods and protocols: Genomic Intelligence Sheetanshu Gupta, Dhirendra Kumar, Radhika Negi, Ranjan Singh, Mohammad Javed Ansari, Shakuli Kashyap, Sudhir Mehrotra, 2024-12-06 The field of metagenomics has revolutionized our comprehension of microbial diversity and function across various habitats, from the human body to terrestrial and aquatic environments. Simultaneously, advancements in AI have empowered researchers to analyze vast troves of genomic data with unprecedented speed and precision, facilitating new insights into the complex interplay between microorganisms and their surroundings. The subject matter in this book provides an overview of metagenomics and discusses the combination of metagenomics and AI and its significant consequences for advancements in science. The chapters examine the approaches, difficulties, and revolutionary uses of AI in metagenomics and provide insight into the convergence of genomics, metagenomics, and AI’s potential to revolutionize diverse fields from healthcare to environmental. Print edition not for sale in South Asia (India, Sri Lanka, Nepal, Bangladesh, Pakistan or Bhutan) |
Enzyme - Wikipedia
An enzyme (/ ˈ ɛ n z aɪ m /) is a protein that acts as a biological catalyst by accelerating chemical reactions. The molecules upon which enzymes may act are called substrates, and the enzyme converts …
Enzyme | Definition, Mechanisms, & Nomenclature …
May 20, 2025 · Enzyme, a catalyst that regulates the rate at which chemical reactions proceed in living organisms without itself being altered in the process. Most critically, enzymes catalyze all …
Enzymes: Function, definition, and examples - Medical News …
Dec 8, 2023 · Enzymes provide help with facilitating chemical reactions within each cell. Since they are not destroyed during the process, a cell can reuse each enzyme repeatedly. This article...
Enzymes: What Are Enzymes, Pancreas, Digestion & Liver Fu…
May 12, 2021 · Enzymes are proteins that help speed up chemical reactions in our bodies. Enzymes are essential for digestion, liver function and much more. Too much or too little of a certain …
Enzymes: principles and biotechnological applications
Enzymes are biological catalysts (also known as biocatalysts) that speed up biochemical reactions in living organisms. They can also be extracted from cells and then used to catalyse a wide range of …
Enzyme - Wikipedia
An enzyme (/ ˈ ɛ n z aɪ m /) is a protein that acts as a biological catalyst by accelerating chemical reactions. The …
Enzyme | Definition, Mechanisms, & Nomenclatur…
May 20, 2025 · Enzyme, a catalyst that regulates the rate at which chemical reactions proceed in living …
Enzymes: Function, definition, and examples - Medical News …
Dec 8, 2023 · Enzymes provide help with facilitating chemical reactions within each cell. Since they are not …
Enzymes: What Are Enzymes, Pancreas, Digestion & Liver F…
May 12, 2021 · Enzymes are proteins that help speed up chemical reactions in our bodies. Enzymes are essential …
Enzymes: principles and biotechnological applications …
Enzymes are biological catalysts (also known as biocatalysts) that speed up biochemical reactions in living …
