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Author: Frances H. Arnold
Publisher: Springer Science & Business Media
ISBN: 1592593968
Category : Science
Languages : en
Pages : 381
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Book Description
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.
Author: Frances H. Arnold
Publisher: Springer Science & Business Media
ISBN: 1592593968
Category : Science
Languages : en
Pages : 381
Get Book Here
Book Description
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.
Author: Frances H. Arnold
Publisher: Humana Press
ISBN: 9781617374722
Category : Science
Languages : en
Pages : 0
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Book Description
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.
Author: Frances H. Arnold
Publisher: Humana Press
ISBN: 9781588292865
Category : Science
Languages : en
Pages : 383
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Book Description
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.
Author:
Publisher:
ISBN:
Category : Combinatorial chemistry
Languages : en
Pages :
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Book Description
Natural selection created optimal catalysts. However, optimal performance of enzyme catalysis does not necessarily refer to maximum reaction rate. Rather, it may be a compromise between specificity, rate, stability, and other chemical constraints that makes enzymes capable of catalyzing reactions under mild conditions and with high substrate specificity, accompanied by high regio- and enantioselectivity. The book presented here focuses on the directed evolution of proteins, which has established itself as a powerful method for designing enzymes showing new substrate specificities. It includes a comprehensive repertoire of techniques for producing combinatorial enzyme libraries, while the functional gene expression in a suitable host helps in selecting the appropriate structure, making fast screening a necessity.
Author: Aaron M. Leconte
Publisher: American Chemical Society
ISBN: 0841296375
Category : Science
Languages : en
Pages : 168
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Book Description
While the field of directed evolution is vast and varied, it comes from a simple principle: to apply the principles of biological evolution in the laboratory to generate useful molecules. This widely used technique has yielded blockbuster drugs, essential biological tools, and many more useful chemicals. Considering its impact, it’s unsurprising that the 2018 Chemistry Nobel Prize was awarded for the use of directed evolution of proteins. Directed Evolution is an excellent starting point for any scientist interested in directed evolution. This primer aims to provide readers with the foundational knowledge and vocabulary that enables them to tackle the literature and begin designing their projects. Directed evolution, as a field, can be challenging to start in; one of the biggest challenges to starting work in this field is that it requires a different style of thinking than many areas of chemistry. CHAPTER 1 explains this different mentality and then gives an overview of directed evolution: what it is, why it is useful, how evolution is applied in the lab, and how evolution can be applied for human benefit. Given the diversity and quantity of methods and approaches in directed evolution, the remaining chapters focus on helping the readers find the right approach to their specific problem by outlining the basic thought process and classical methods in directed evolution. Specifically, CHAPTER 2 discusses applying directed evolution to nucleic acids. Nucleic acid evolution has several unique features that make it easier to understand conceptually, making it a good starting point for understanding how to think about and leverage evolution. CHAPTERS 3 through 6 cover approaches to evolving proteins. CHAPTER 3 discusses the strengths and weaknesses of different approaches to the diversification of proteins, giving the reader a framework for designing libraries. CHAPTERS 4, 5, and 6 introduce different approaches to sorting by applying selection pressures to find the best variant from a large complex library. Upon completing this primer, the reader will have a theoretical basis to design and execute a directed evolution campaign. They will have the core knowledge to dive into the larger world of directed evolution and begin designing projects that leverage this powerful technique.
Author: Huimin Zhao
Publisher: John Wiley & Sons
ISBN: 3527344705
Category : Science
Languages : en
Pages : 41
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Book Description
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.
Author: Miguel Alcalde
Publisher: Springer
ISBN: 3319504134
Category : Science
Languages : en
Pages : 286
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Book Description
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.
Author: National Academy of Sciences
Publisher:
ISBN:
Category : Science
Languages : en
Pages : 388
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Book Description
The Arthur M. Sackler Colloquia of the National Academy of Sciences address scientific topics of broad and current interest, cutting across the boundaries of traditional disciplines. Each year, four or five such colloquia are scheduled, typically two days in length and international in scope. Colloquia are organized by a member of the Academy, often with the assistance of an organizing committee, and feature presentations by leading scientists in the field and discussions with a hundred or more researchers with an interest in the topic. Colloquia presentations are recorded and posted on the National Academy of Sciences Sackler colloquia website and published on CD-ROM. These Colloquia are made possible by a generous gift from Mrs. Jill Sackler, in memory of her husband, Arthur M. Sackler.
Author: Susanne Brakmann
Publisher: Wiley-VCH
ISBN: 9783527304233
Category : Science
Languages : en
Pages : 0
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Book Description
Die natürliche Auslese hat optimale Protein-Katalysatoren hervorgebracht; eine optimale Enzymkatalyse indes verläuft nicht notwendigerweise mit der höchstmöglichen Geschwindigkeit. Vielmehr ist es oft ein Kompromiß zwischen Spezifität, Geschwindigkeit, Stabilität und vielen anderen Faktoren, den die Natur für eine chemische Reaktion unter milden Bedingungen, mit hoher Regio- und Enantioselektivität, eingeht. Vor diesem Hintergrund hat sich die gerichtete Evolution von Proteinen als eine leistungsfähige Methode für das Design von Enzymen mit neuen Substratspezifitäten entwickelt. Das vorliegende Buch beleuchtet das umfassende Repertoire an Techniken für das Erstellen von kombinatorischen Enzymbibliotheken, angefangen von den theoretischen Grundlagen bis hin zur praktischen Anwendung - die immer mehr auch in der klassischen organischen Synthese an Bedeutung gewinnt!
Author: Frances H. Arnold
Publisher: Humana Press
ISBN: 9781617374715
Category : Science
Languages : en
Pages : 0
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Book Description
Biological systems are very special substrates for engineering—uniquely the products of evolution, they are easily redesigned by similar approaches. A simple algorithm of iterative cycles of diversification and selection, evolution works at all scales, from single molecules to whole ecosystems. In the little more than a decade since the first reported applications of evolutionary design to enzyme engineering, directed evolution has matured to the point where it now represents the centerpiece of industrial biocatalyst development and is being practiced by thousands of academic and industrial scientists in com- nies and universities around the world. The appeal of directed evolution is easy to understand: it is conceptually straightforward, it can be practiced without any special instrumentation and, most important, it frequently yields useful solutions, many of which are totally unanticipated. Directed evolution has r- dered protein engineering readily accessible to a broad audience of scientists and engineers who wish to tailor a myriad of protein properties, including th- mal and solvent stability, enzyme selectivity, specific activity, protease s- ceptibility, allosteric control of protein function, ligand binding, transcriptional activation, and solubility. Furthermore, the range of applications has expanded to the engineering of more complex functions such as those performed by m- tiple proteins acting in concert (in biosynthetic pathways) or as part of mac- molecular complexes and biological networks.
