Directed Evolution of Selective Enzymes

Author: Manfred T. Reetz
Publisher: John Wiley & Sons
ISBN: 3527316604
Format: PDF, Kindle
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Authored by one of the world's leading organic chemists, this authoritative reference provides an overview of basic strategies in directed evolution and introduces common gene mutagenesis, screening and selection methods. Throughout the text, emphasis is placed on methodology development to maximize efficiency, reliability and speed of the experiments and to provide guidelines for efficient protein engineering. Professor Reetz highlights the application of directed evolution experiments to address limitations in the field of enzyme selectivity, substrate scope, activity and robustness. He critically reviews recent developments and case studies, takes a look at future applications in the field of organic synthesis, and concludes with lessons learned from previous experiments.

Directed Enzyme Evolution

Author: Frances Hamilton Arnold
Publisher: Springer Science & Business Media
ISBN: 1592593968
Format: PDF, Kindle
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Seasoned practitioners from many leading laboratories describe their best readily reproducible screening strategies for isolating useful clones. These techniques have been optimized for sensitivity, high throughput, and robustness, and are of proven utility for directed evolution purposes. The assays presented use a variety of techniques, including genetic complementation, microtiter plates, solid-phase screens with colorimetric substrates, and flow cytometric screens. An accompanying volume, Directed Evolution Library Creation: Methods and Protocols, describes readily reproducible methods for the creation of mutated DNA molecules and DNA libraries.

Biocatalysis and Agricultural Biotechnology

Author: Ching T. Hou
Publisher: CRC Press
ISBN: 9781420077070
Format: PDF, ePub
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Worldwide energy and food crises are spotlighting the importance of bio-based products – an area many are calling on for solutions to these shortages. Biocatalysis and Agricultural Biotechnology encapsulates the cutting-edge advances in the field with contributions from more than 50 international experts comprising sectors of academia, industry, and government research institutes, a virtual Who’s Who among biocatalysis scientists. Created Under the Editorial Guidance of Leading Biotechnology Experts With the aid of numerous graphs and illustrations, this authoritative reference documents such important advances as: Cloning and characterization of Kennedy pathway acyltransferases Engineering of plants for industrial uses New approaches from acquired tolerance to the biotic and abiotic stress of economically important crops This comprehensive text also explores a variety of bio-based industrial products, including: The modification of enzyme character through gene manipulation The biocatalytic synthesis of chiral intermediates for drug development The use of Omega-3 phospholipid nano capsules as effective forms for transporting immune response modifiers Providing in-depth reviews of this ancient field and its modern-day advances, Biocatalysis and Agricultural Biotechnology is an invaluable lab reference for teachers, graduate students, and industrial scientists conducting research in the biosciences.

Organotitanium Reagents in Organic Synthesis

Author: Manfred Reetz
Publisher: Springer Science & Business Media
ISBN: 3642707041
Format: PDF, ePub, Docs
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Titanium has been used to perform many kinds of reactions in organic and inorganic chemistry. The present book is concerned primarily with a new development in titanium chemistry which is useful in organic synthesis. In 1979/80 it was discovered that the titanation of classical carbanions using C1TiX leads to species with reduced basicity and reactivity. This increases 3 chemo-, regio-and stereo selectivity in reactions with organic compounds such as aldehydes, ketones and alkyl halides. Many new examples have been reported in recent times. Since the nature of the ligand X at titanium can be widely varied, the electronic and steric nature of the reagents is easily controlled. This helps in predicting the stereochemical outcome of many of the C-C bond forming reactions, but the trial and error method is still necessary in other cases. One of the ultimate objectives of chemistry is to understand correlations between structure and reactivity. Although this goal has not been reached in the area of organotitanium chemistry, appreciable progress has been made. A great deal of physical and computational data of organotitanium compounds described in the current and older literature (e. g. , Ziegler-Natta type catalysts) has been reported by polymer, inorganic and theoretical chemists. It is summarized in Chapter 2 of this book, because some aspects are useful in understanding reactivity and selectivity of organo titanium compounds in organic synthesis as described in the chapters which follow.

Biocatalysis in the Pharmaceutical and Biotechnology Industries

Author: Ramesh N. Patel
Publisher: CRC Press
ISBN: 1420019376
Format: PDF, Kindle
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Because enzyme-catalyzed reactions exhibit higher enantioselectivity, regioselectivity, substrate specificity, and stability, they require mild conditions to react while prompting higher reaction efficiency and product yields. Biocatalysis in the Pharmaceutical and Biotechnology Industries examines the use of catalysts to produce fine chemicals and chiral intermediates in a variety of pharmaceutical, agrochemical, and other biotechnological applications. Written by internationally recognized scientists in biocatalysis, the authors analyze the synthesis of chiral intermediates for over 60 brand-name pharmaceuticals for a wide range of drug therapies and treatments. From starting material to product, the chapters offer detailed mechanisms that show chiral intermediates and other by-products for each reaction—including hydrolytic, acylation, halogenation, esterification, dehalogenation, oxidation-reduction, oxygenation, hydroxylation, deamination, transamination, and C–C, C–N, C–O bonds formation. Cutting-edge topics include advanced methodologies for gene shuffling and directed evolution of biocatalysts; the custom engineering of enzymes; the use of microbial cells and isolated biocatalysts; the use of renewable starting materials; and generating novel molecules by combinatorial biocatalysis and high-throughput screening. Focusing on industrial applications, the book also considers factors such as bulk processes, instrumentation, solvent selection, and techniques for catalyst immobilization, reusability, and yield optimization throughout. Biocatalysis in the Pharmaceutical and Biotechnology Industries showcases the practical advantages and methodologies for using biocatalysts to develop and produce chiral pharmaceuticals and fine chemicals.

Directed Enzyme Evolution Advances and Applications

Author: Miguel Alcalde
Publisher: Springer
ISBN: 3319504134
Format: PDF, Docs
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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.

Chemical Aspects of Enzyme Biotechnology

Author: Thomas O. Baldwin
Publisher: Springer Science & Business Media
ISBN: 1475796374
Format: PDF, ePub, Mobi
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The Industry-University Cooperative Chemistry Program has sponsored seven previous international symposia covering a wide variety of topics of interest to industrial and academic chemists. The eighth IUCCP symposium, held March 19-22, 1990, at Texas A&M University, represents a deviation from the former symposia, in that it is the first of a two-symposium series dedicated to the rapidly moving new field of industrial biochemistry that has beco~e known as biotechnology. Biotechnology is really not a new discipline, but rather is a term coined to describe the new and exciting commercial applications of biochemistry. The development of the field of biotechnology is a direct result of recombinant DNA technology, which began in earnest about 15 years ago. Today, we can routinely do experiments that were inconceivable in the early 1970's. Only comparatively simple technology available even in small laboratories is required to synthesize a gene and from it, to produce vast amounts of biological materials of enormous commercial value. These technical developments and others have stimulated increased activities in the field of enzyme biotechnology, using enzymes to catalyze "unnatural" reactions to produce complex molecules with stereochemical precision. It is true today, we can readily produce DNA fragments that will encode any amino acid sequence that we might desire, but at this point, our foundation of basic knowledge falls short. The dream of "designer enzymes" is still a fantasy, but the current wave of research activity and exciting new developments suggest that in the future the dream may become a reality.

Biocatalysis for the Pharmaceutical Industry

Author: Junhua (Alex) Tao
Publisher: John Wiley & Sons
ISBN: 0470823151
Format: PDF, ePub
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Biocatalysis is rapidly evolving into a key technology for the discovery and production of chemicals, especially in the pharmaceutical industry, where high yielding chemo-, regio-, and enantioselective reactions are critical. Taking the latest breakthroughs in genomics and proteomics into consideration, Biocatalysis for the Pharmaceutical Industry concisely yet comprehensively discusses the modern application of biocatalysis to drug discovery, development, and manufacturing. Written by a team of leading experts, the book offers deep insight into this cutting edge field. Covers a wide range of topics in a systematic manner with an emphasis on industrial applications Provides a thorough introduction to the latest biocatalysts, modern expression hosts, state-of-the-art directed evolution, high throughput screening, and bioprocess engineering Addresses frontier subjects such as emerging enzymes, metabolite profiling, combinatorial biosynthesis, metabolic engineering, and autonomous enzymes for the synthesis and development of chiral molecules, drug metabolites, and semi-synthetic medicinal compounds and natural product analogs Highlights the impact of biocatalysis on green chemistry Contains numerous graphics to illustrate concepts and techniques Biocatalysis for the Pharmaceutical Industry is an essential resource for scientists, engineers, and R&D policy makers in the fine chemical, pharmaceutical, and biotech industries. It is also an invaluable tool for academic researchers and advanced students of organic and materials synthesis, chemical biology, and medicinal chemistry.

Enzymes in Synthetic Organic Chemistry

Author: Chi-Huey Wong
Publisher: Elsevier
ISBN: 9780080359410
Format: PDF
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This book covers the most recent development of enzymatic organic synthesis, with particular focus on the use of isolated enzymes. It is organized into one introductory chapter dealing with the characteristics of enzymes as catalysts, and five chapters dealing with different types of chemical transformations. Methods for enzyme immobilization and stabilizaton, the use of enzymes in extreme environments, and the alteration of enzyme properties by chemical modification and site-directed mutagenesis for synthetic purposes are covered.