Directed Enzyme Evolution Advances And Applications
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Directed Enzyme Evolution: Advances and Applications
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.
Directed Enzyme Evolution
Author: Frances H. Arnold
language: en
Publisher: Springer Science & Business Media
Release Date: 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.
The Autotrophic Biorefinery
Author: Robert Kourist
language: en
Publisher: Walter de Gruyter GmbH & Co KG
Release Date: 2021-10-25
The depletion of fossil resources and an ever-growing human population create an increasing demand for the development of sustainable processes for the utilization of renewable resources. As autotrophic microorganisms offer numerous metabolic pathways for the fixation of carbon dioxide and the metabolic utilization of light, electricity and inorganic energy donors, they are expected to play a pivotal role in an emerging carbon neutral society. This text-book presents the metabolic principles of autotrophy and current efforts for their utilization in biotechnology, including photoautotrophic, chemolithoautotrophic and electroautotrophic organisms. It outlines how modern molecular biology and process engineering create technologies that allow to use industrial off-gases and inorganic energy for the synthesis of bio-based plastics, materials and other chemical products. The text-book is ideally suited for students in advanced graduate and master courses and offers a reference for PhD students, engineers, chemists, biologists and all with an interests in biotechnology and renewable resources.